RU2777911C2 - Combination of cell therapy and immunomodulator compound - Google Patents

Abstract

FIELD: immunotherapy.

SUBSTANCE: treatment method includes the administration of T-cell therapy to a subject with cancer, where T-cell therapy includes genetically engineered T-lymphocytes expressing a chimeric antigen receptor (CAR), which is specifically binds to an antigen related to cancer; and the administration to a subject of an immunomodulator compound, which relieves degradation of Ikaros (IKZF1) and/or Aiolos (IKZF3), where the beginning of the administration of the specified immunomodulator compound is performed at least two days after the beginning of the administration of T-cell therapy. Another option of the treatment method is also disclosed.

EFFECT: group of inventions provides for increased or improved activity, efficiency, persistency, reproduction and/or proliferation of T-lymphocytes for adoptive cell therapy or endogenous T-lymphocytes recruited by immunotherapeutic agents.

80 cl, 40 dwg, 6 tbl, 18 ex

Description

Cross-references to related applications

[0001] This application claims priority of U.S. Provisional Application No. 62/492,947, filed May 1, 2017, entitled "COMBINATION OF A CELL THERAPY AND AN IMMUNOMODULATORY COMPOUND", U.S. Provisional Application No. 62/538,670, filed July 29, 2017 entitled “COMBINATION OF A CELL THERAPY AND AN IMMUNOMODULATORY COMPOUND”, U.S. Provisional Application No. 62/549390, filed August 23, 2017, entitled “COMBINATION OF A CELL THERAPY AND AN IMMUNOMODULATORY COMPOUND”, U.S. Provisional Application No. 62/580433, filed November 1, 2017, entitled “COMBINATION OF A CELL THERAPY AND AN IMMUNOMODULATORY COMPOUND”, and U.S. Provisional Application No. 62/596753, filed December 8, 2017, entitled “COMBINATION OF A CELL THERAPY AND AN IMMUNOMODULATORY COMPOUND”, the contents of which are incorporated by reference in their entirety.

Inclusion as a reference of Sequence Listing

[0002] This application is filed with the Sequence Listing in electronic format. The sequence list is offered as a file named 735042009640SeqList.TXT, created on April 30, 2018, which has a size of 328355 bits. Information in the electronic format of the Sequence Listing is incorporated by reference in its entirety.

The field of technology to which the invention relates

[0003] The present invention in some aspects relates to methods, compositions and uses, including immunotherapy, such as adoptive cell therapy, for example, T-cell therapy, and an immunomodulatory compound, such as a structural or functional analog or derivative of thalidomide and / or an E3 ubiquitin ligase inhibitor. The provided methods, compositions, and uses include methods, compositions, and uses for combination therapy involving the administration or application of one or more immunomodulatory compounds in combination with T cell therapy, such as genetically engineered T lymphocyte therapy, including cells with recombinant receptors. , for example, using T-lymphocytes expressing chimeric antigen receptors (CARs). Also provided are compositions, methods of administration to subjects, articles of manufacture, and kits for use in the methods. In some aspects, the features of the methods and cells provide increased or improved activity, efficacy, persistence, expansion and/or proliferation of T lymphocytes for adoptive cell therapy or endogenous T lymphocytes recruited with immunotherapeutic agents.

State of the art

[0004] Various immunotherapy strategies are available, for example, the introduction of genetically engineered T-lymphocytes for adoptive therapy. For example, strategies are available for genetically engineering T lymphocytes expressing engineered antigen receptors such as CARs and administering compositions containing such cells to subjects. Improved strategies are needed to improve the efficiency of cells, for example, to improve the persistence, activity and/or proliferation of cells when administered to subjects. Methods, compositions, kits, and systems are provided that meet such needs.

The essence of the invention

[0005] This document proposes a combination therapy, including the introduction of immunotherapy, including the functioning or activity T lymphocytes, such as T cell therapy; and an immunomodulatory compound, such as a structural or functional analog or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor. In some aspects, the present methods improve or modulate the proliferation and/or activity or activities of T lymphocytes associated with the administration of immunotherapy or an immunotherapeutic agent, such as a composition containing cells for adoptive cell therapy, such as, for example, T cell therapy (for example, CAR-expressing T-lymphocytes). In some embodiments, combination therapy typically comprises administering an immunomodulatory compound, such as a structural or functional analog of thalidomide and/or an E3 ubiquitin ligase inhibitor (e.g., lenalidomide (3-(4-amino-1-oxo-1,3- dihydro-2H-isoindol-2-yl)piperidine-2,6-dione)) and administering a T cell therapy, such as a composition containing cells for adoptive cell therapy, such as, for example, T cell therapy (e.g., CAR expressing T-lymphocytes).

[0006] Provided herein are methods of treatment that include: (a) administering T cell therapy to a subject having a disease or condition; and (b) administering to the subject an immunomodulatory compound.

[0007] Provided herein are methods of treatment that include administering T cell therapy to a subject having a disease or condition, wherein at the time the T cell therapy is started, the subject is being administered and/or being treated with an immunomodulatory compound, and/ or a blood sample or biopsy of the subject contains detectable levels of T-lymphocytes from therapy with genetically engineered T-lymphocytes.

[0008] Provided herein are methods of treatment that include administering an immunomodulatory compound to a subject having a disease or condition where, at the time the immunomodulatory compound is started, the subject has previously received T cell therapy for the treatment of the disease or condition, and/or a blood sample or a biopsy of the subject contains detectable levels of T-lymphocytes from therapy with genetically engineered T-lymphocytes. In some embodiments, the method thereby prevents, reduces, or alleviates one or more symptoms or outcomes of diseases or conditions.

[0009] In some embodiments of any of the methods provided herein, (a) the amount of immunomodulatory compound administered is insufficient, as a single agent and/or without the administration of T cell therapy, to alleviate, reduce, or prevent the disease or condition, or symptom or result; and/or (b) the amount of the immunomodulatory compound administered is insufficient, as a single agent and/or without the administration of T cell therapy, to alleviate, reduce, or prevent the subject's disease or condition or symptom or result; and/or (c) thereby the method reduces or alleviates the symptom or outcome or severity disease or condition to a degree that is greater than a combination of (i) the degree of reduction or amelioration effected by the administration of the immunomodulatory agent alone, optionally the average of this indicator for a population of subjects having the disease or condition, and (ii) the degree of reduction or amelioration by the administration one only T-cell therapy, optionally, the average value of this indicator for a population of subjects with a disease or condition; and/or (d) the amount of the immunomodulatory compound administered in the method, or the compound administered in one or more doses, is a maintenance dose of the compound or corresponds to the dose of the compound administered to subjects showing a response, optionally a complete response, following administration of the compound for treatment.

[0010] In some embodiments of any of the methods provided herein, the disease or condition is untreatable or resistant to an immunomodulatory compound and/or becomes untreatable or resistant thereto after treatment with an immunomodulatory compound; and/or the subject or disease or condition is determined to have a mutation or factor that renders the disease or condition resistant to treatment with an immunomodulatory compound.

[0011] In some embodiments of any of the methods provided herein, the immunomodulatory compound is selected from: immunomodulatory drugs (IMiDs), thalidomide analogs, thalidomide derivatives, compounds that interact and/or bind to cereblon (CRBN) and/or from one or more elements of the CRBN and E3 ubiquitin ligase complex, Ikaros (IKZF1) inhibitors, Aiolos (IKZF3) inhibitors, compounds that enhance or facilitate the ubiquitination and/or degradation of Ikaros (IKZF1) and/or Aiolos (IKZF3).

[0012] Provided herein are methods of treatment that include (a) administering T cell therapy to a subject having a disease or condition; and (b) administering to the subject an immunomodulatory compound, wherein said immunomodulatory compound is selected from the group consisting of lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2, 6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione) or avadomide (3-(5-amino-2-methyl-4-oxo- 4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomers, enantiomers or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

[0013] Provided herein are methods of treatment that include administering T cell therapy to a subject having a disease or condition, wherein at the time the T cell therapy is started, the subject is being administered and/or being treated with an immunomodulatory compound, and/ or a blood sample or biopsy of the subject contains detectable levels of T-lymphocytes from therapy with genetically engineered T-lymphocytes, where the specified immunomodulatory compound is selected from the group consisting of lenalidomide (3-(4-amino-1-oxo-1,3- dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione) or avadomide (3 -(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomers, enantiomers or mixture of enantiomers, or their pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph.

[0014] Provided herein are methods of treatment that include administering an immunomodulatory compound to a subject having a disease or condition where, at the time the immunomodulatory compound is administered, the subject has previously received T cell therapy for the treatment of the disease or condition, and/or a blood sample or the subject's biopsy contains detectable levels of T lymphocytes from genetically engineered T lymphocyte therapy, wherein said immunomodulatory compound is selected from the group consisting of lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H- isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione) or avadomide (3-(5- amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomers, enantiomers or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph.

[0015] Provided herein are methods of treatment that include (a) administering T cell therapy to a subject having a disease or condition; and (b) administering to the subject an immunomodulatory compound, wherein said immunomodulatory compound is selected from the group consisting of lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2, 6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione) or avadomide (3-(5-amino-2-methyl-4-oxo- 4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomers, enantiomers, or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, and where the start of administration of the immunomodulatory compound corresponds to the time time: (1) at least 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the start of T- cell therapy, and/or it is carried out after 2-28 or 7-21 days after the start of the introduction of T-cell therapy; and/or (2) at or after the time point, optionally immediately after, or within 1-3 days after: (i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time points after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0016] Provided herein are methods of treatment that include administering an immunomodulatory compound to a subject who is administered prior to initiation of administration of the immunomodulatory compound, T cell therapy for treating a disease or condition, wherein said immunomodulatory compound is selected from the group consisting of lenalidomide (3-( 4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxpiperidin-3-yl) isoindole-1,3-dione) or avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomer, enantiomer or mixture enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, and wherein the start of administration of the immunomodulatory compound corresponds to the time point: (1) at least 2 days, at least 1 week, at least , 2 weeks, at least 3 weeks, or at least 4 weeks after the start of T-cell administration therapy, and/or it is carried out after 2-28 or 7-21 days after the start of the introduction of T-cell therapy; and/or (2) at or after a time point, optionally immediately thereafter or within 1-3 days after: (i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0017] Provided herein are methods of treatment that comprise administering a therapeutically effective amount of an immunomodulatory compound, wherein said immunomodulatory compound is selected from the group consisting of lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H- isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione) or avadomide (3-(5- amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomers, enantiomers or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph, to a subject who, prior to the start of administration of an immunomodulatory compound, is administered T-cell therapy for the treatment of a disease or condition, where the subject is such that on or about day 12-15, optionally on or about day 14, after the initiation of T cell therapy for the treatment of a disease or condition: (i) the number of cells from T cells less than 75% of the average number of cells from T-cell therapy in a subject at the same time in a plurality of subjects who are administered the same or similar dose of T-cell therapy; and/or (ii) the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, less than 10 cells per µl, less than 5 cells per µl, or less than 1 cell per µl in the blood.

[0018] Provided herein are methods of treatment that include (a) selecting a subject that corresponds to or approximately corresponds to day 12-15, optionally, day 14 or so, after the start of the administration of T cell therapy for the treatment of a disease or condition: (i) the number of cells from T-cell therapy in the subject is less than 75% of the average number of cells from T-cell therapy at the same time in a plurality of subjects who are administered the same or similar dose of T-cell therapy; and/or (ii) the number of CD3+ or CD8+ cells from T-cell therapy, optionally CAR+ T-lymphocytes, in the blood is less than 10 cells per µl, less than 5 cells per µl, or less than 1 cell per µl; and (b) administering a therapeutically effective amount of an immunomodulatory compound to a subject, wherein said immunomodulatory compound is selected from the group consisting of lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine -2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione) or avadomide (3-(5-amino-2-methyl-4 -oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomer, enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

[0019] Provided herein are methods of treatment that include administering T cell therapy to a subject having a disease or condition, wherein the subject is administered an immunomodulatory compound prior to initiation of T cell therapy, wherein said immunomodulatory compound is selected from the group consisting of: lenalidomide (3 -(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxpiperidin-3- yl)isoindol-1,3-dione) or avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomer, enantiomer or a mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, and wherein the immunomodulatory compound is administered in a cycle comprising: (i) administration for up to 21 days, sequentially, where the cycle comprises more than 30 days from start administration of an immunomodulatory compound; and/or (ii) administration for multiple consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or (iii) administration for no more than 14 consecutive days.

[0020] Provided herein are methods of treatment that include administering an immunomodulatory compound to a subject, wherein the subject has a disease or condition and is being administered T cell therapy, wherein said immunomodulatory compound is selected from the group consisting of: lenalidomide (3-(4-amino -1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1 ,3-dione) or avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomer, enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, and wherein the immunomodulatory compound is administered in a cycle comprising: (i) administering the immunomodulatory compound for up to 21 days, consecutively, wherein the cycle comprises more than 30 days from the start of administration of the immunomodulatory compound ; and/or (ii) administering an immunomodulatory compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or (iii) administering an immunomodulatory compound for up to 14 consecutive days.

[0021] In some embodiments of any of the methods provided herein, administering an immunomodulatory compound comprises: (i) at least one cycle greater than 30 days from the start of administration of the immunomodulatory compound, wherein the cycle comprises administering the compound, optionally daily, or, at least daily, for up to 21 days, consecutively, and/or where the last administration of the compound in the cycle corresponds to 21 days or less after the first administration of the compound in the cycle; and/or (ii) at least two cycles, every at least two cycles comprising administering the compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or (iii) administration, optionally daily, or at least daily, for no more than 14 consecutive days.

[0022] In some embodiments of any of the methods provided herein, the start of the administration of the immunomodulatory compound or the start of the administration of the compound in at least one cycle, and the start of the administration of T-cell therapy occurs on the same day or in consecutive days, optionally, simultaneously; and/or at least one dose of the immunomodulatory compound is administered on the same day, within one or two days, before or after a dose of T cell therapy.

[0023] In some embodiments of any of the methods provided herein, the initiation of administration of the immunomodulatory compound, or the initiation of administration of the compound in at least one cycle, occurs prior to the initiation of administration of T cell therapy.

[0024] Provided herein are methods of treatment that include administering T cell therapy to a subject having a disease or condition, wherein the subject is administered an immunomodulatory compound prior to initiation of T cell therapy, wherein the cycle comprises: (i) administering for up to 21 days, sequentially, where the cycle includes more than 30 days from the start of the administration of the immunomodulatory compound; and/or (ii) administration for multiple consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or (iii) administration for no more than 14 consecutive days. In some embodiments, the start of administration of the immunomodulatory compound occurs within 14 days prior to the start of T cell therapy.

[0025] In some embodiments provided herein, the immunomodulatory compound is selected from the group consisting of: thalidomide analogs; thalidomide derivatives; compounds that interact and/or bind to cereblon (CRBN) and/or one or more elements of the CRBN and E3 ubiquitin ligase complex; Ikaros inhibitors (IKZF1); inhibitors of Aiolos (IKZF3); and compounds that enhance or facilitate the ubiquitination and/or degradation of Ikaros (IKZF1) and/or Aiolos (IKZF3).

[0026] In some embodiments of any of the methods provided herein, the administration of an immunomodulatory compound begins prior to the administration of T cell therapy, it begins: (i) one week or within one week before or after the collection of a sample containing T from the subject -lymphocytes to be processed and/or genetically engineered to receive therapy, optionally, where the sample is an apheresis sample; and/or (ii) within 14 days prior to initiation of T cell therapy.

[0027] In some embodiments of any of the methods provided herein, the T cell therapy comprises engineered cells to express the recombinant receptor. In some embodiments, the genetic engineering comprises one or more ex vivo preparation method steps, optionally selected from: (1) isolating cells from a biological sample using leukapheresis or apheresis; (2) selecting or enriching cells using immunoaffinity methods; (3) introducing the recombinant nucleic acid, optionally a viral vector, into the cells; (4) incubating cells, optionally genetically engineered cells, in the presence of one or more stimulation conditions; (5) preparing cells in the presence of a cryoprotectant; and/or (6) preparing cells for administration to a subject, optionally in the presence of a pharmaceutically acceptable excipient.

[0028] In some embodiments of any of the methods provided herein, the method includes performing the preparation method and/or further comprising genetically engineering T lymphocytes to express the recombinant receptor, thereby generating a T cell therapy. In some embodiments of any of the methods provided herein, the method comprises contacting cells with an immunomodulatory compound during one or more of the ex vivo preparation steps of the method.

[0029] In some embodiments of any of the methods provided herein, the T cell therapy comprises engineered T lymphocytes obtained by a preparation method comprising ex vivo incubation of the cells in the presence of an immunomodulatory compound.

[0030] In some embodiments of any of the methods provided herein, the method includes incubating cells in the presence of one or more stimulation conditions, which is carried out in the presence of an immunomodulatory compound.

[0031] In some embodiments of any of the methods provided herein, the start of administration of the immunomodulatory compound occurs within 10 days, 7 days, 4 days, 3 days, or 2 days prior to the start of T cell therapy. In some embodiments of any of the methods provided herein, the start of administration of the immunomodulatory compound, at least one cycle, occurs after the start of administration of T cell therapy.

[0032] Provided herein are methods of treatment that include administering an immunomodulatory compound to a subject, wherein the subject has a disease or condition and is being administered T cell therapy, wherein the immunomodulatory compound is administered in a cycle including: (i) administering the immunomodulatory compound for up to 21 days, consecutively, where the cycle includes more than 30 days from the start of the administration of the immunomodulatory compound; and/or (ii) administering an immunomodulatory compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or (iii) administering an immunomodulatory compound for up to 14 consecutive days.

[0033] In some embodiments of any of the methods provided herein, T cell therapy is a therapy in which the peak population cells from therapy, which are optionally CD3+ or CD8+ cells from T-cell therapy, and/or optionally are CAR+ T-lymphocytes, in blood is ((a) the mean of multiple subjects treated with T-cell therapy without administration of an immunomodulatory compound, or (b) subjects after administration of T-cell therapy) is less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl.

[0034] In some embodiments of any of the methods provided herein, the T cell therapy comprises cells expressing a recombinant receptor, optionally a CAR. In some embodiments of any of the methods provided herein, the recombinant receptor comprises a B cell maturation antigen (BCMA) specific antigen-binding domain.

[0035] In some embodiments of any of the methods provided herein, the start of administration of the immunomodulatory compound, at least one cycle, is carried out after the start of the administration of T-cell therapy. In some embodiments of any of the methods provided herein, the start of administration of the immunomodulatory compound is at least 2 days later, at least 1 week later, at least 2 weeks later, at least 3 weeks later or at least 4 weeks after the start of administration or after the last dose of T-cell therapy, and/or it is carried out 2-28 or 7-21 days after the start of administration or after the last dose of T-cell therapy.

[0036] Provided herein are methods of treatment that include (a) administering T cell therapy to a subject having a disease or condition; and (b) administering to the subject an immunomodulatory compound, wherein the start of administration of the immunomodulatory compound corresponds to the time point: (a) at least 2 days later, at least 1 week later, at least 2 weeks later, at least 3 weeks or at least 4 weeks after the start of the introduction of T-cell therapy, and/or it is carried out after 2-28 or 7-21 days after the start of the introduction of T-cell therapy; and/or (b) at or after a time point, optionally immediately after or within 1-3 days after: (i) a peak maximum cell level from T cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0037] Provided herein are methods of treatment that include administering an immunomodulatory compound to a subject who is receiving T cell therapy prior to initiation of administration of the immunomodulatory compound for the treatment of a disease or condition, where the onset of administration of the immunomodulatory compound corresponds to a time point of: (a) at least after 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the start of the introduction of T-cell therapy, and / or it is carried out through 2-28 or 7-21 days after the start of the introduction of T-cell therapy; and/or (b) at or after a time point, optionally immediately after or within 1-3 days after: (i) a peak peak cell level from T cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0038] In some embodiments of any of the methods provided herein, the start of administration of the immunomodulatory compound is after a period of time that is greater than about 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 24 days, or 28 days after initiation of T-cell therapy.

[0039] In some embodiments of any of the methods provided herein, prior to initiation of administration of an immunomodulatory compound, a subject is selected in which: (i) a peak maximum cell level from T cell therapy is detected in the blood; (ii) the number of cells from T-cell therapy detected in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared to the previous point in time after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0040] Provided herein are methods of treatment that include administering a therapeutically effective amount of an immunomodulatory compound to a subject who, prior to initiation of administration of the immunomodulatory compound, is receiving T-cell therapy for the treatment of a disease or condition, wherein the subject is one who on day 12-15 or on or about that day, optionally on or about day 14, after initiation of T cell therapy for the treatment of a disease or condition: (i) the subject's T cell count is less than 75% of the mean T cell count -cell therapy at the same time in multiple subjects who are administered the same or similar dose of T-cell therapy; and/or (ii) the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, in the blood is less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl.

[0041] Provided herein are methods of treatment that include (a) selecting a subject that corresponds to, or approximately corresponds to, day 12-15, optionally, day 14 or so, after initiation of T cell therapy for the treatment of a disease or condition: (i) the number of cells from T-cell therapy in the subject is less than 75% of the average number of cells from T-cell therapy at the same time in a plurality of subjects who are administered the same or similar dose of T-cell therapy; and/or (ii) the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, in the blood is less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl; and (b) administering a therapeutically effective amount of the immunomodulatory compound to the subject. In some embodiments of any of the methods provided herein, the immunomodulatory compound is administered daily, optionally once daily.

[0042] In some embodiments of any of the methods provided herein, the immunomodulatory compound is administered for more than or about more than 7 consecutive days, more than about more than about 14 consecutive days, more than or about more than about 21 consecutive days, more than or about more than about 21 consecutive days. consecutive day, or more, or more than about 28 consecutive days. In some embodiments of any of the methods provided herein, the immunomodulatory compound is administered in a cycle comprising daily administration for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered. In some embodiments, the rest period during which no immunomodulatory compound is administered is greater than 7 consecutive days, greater than 14 consecutive days, greater than 21 days, or greater than 28 days.

[0043] In some embodiments of any of the methods provided herein, the cycle of administration of the immunomodulatory compound is repeated at least once. In some embodiments, the immunomodulatory compound is administered for at least 2 cycles, at least 3 cycles, at least 4 cycles, at least 5 cycles, at least 6 cycles, at least 7 cycles, at least 8 cycles, at least 9 cycles, at least 10 cycles, at least 11 cycles, or at least 12 cycles.

[0044] In some embodiments of any of the methods provided herein, the administration of the immunomodulatory compound continues at least from the time after the start of the administration of T-lymphocytes, until: the number of cells from the administered T-cell therapy or received of those detected in the subject's blood will not increase compared to the subject's cell count at the time immediately prior to the administration of the immunomodulatory compound or compared to the time prior to the administration of the T-cell therapy; the number of cells from or derived from T-cell therapy detected in the blood will not be within 2.0 times (more or less) the peak or maximum number observed in the blood of the subject after the start of the administration of T-lymphocytes; the number of cells from T-cell therapy detected in the subject's blood will not become greater than or about greater than 10%, 15%, 20%, 30%, 40%, 50%, or 60% of the total number of peripheral blood mononuclear cells (PBMC) in subject's blood; and/or the subject will not show a reduction in tumor weight relative to tumor weight at the time immediately prior to administration of the T cell therapy or at the time immediately prior to administration of the immunomodulatory compound; and/or the subject will not demonstrate complete or clinical remission.

[0045] In some embodiments of any of the methods provided herein, the immunomodulatory compound binds to Cereblon (CRBN) and/or to the CRBN and E3 ubiquitin ligase complex; and/or inhibits the transcription growth factor Ikaros (IKZF1) or Aiolos (IKZF3); and/or enhances ubiquitination or degradation of Ikaros (IKZF1) or Aiolos (IKZF3).

[0046] In some embodiments of any of the methods provided herein, the immunomodulatory compound is thalidomide or is a derivative or analog of thalidomide. In some embodiments, the immunomodulatory compound is lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione) or pomalidomide (4-amino -2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione), avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine -2,6-dione), stereoisomer of lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione), pomalidomide (4-amino -2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione), avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine -2,6-dione) or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In some embodiments, the immunomodulatory compound is lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione), a stereoisomer of lenalidomide (3- (4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione) or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

[0047] In some embodiments, provided herein, the immunomodulatory compound is lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione ), pomalidomide (4-amino-2-(2,6-dioxo-piperidin-3-yl)isoindol-1,3-dione) or avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazoline -3-yl)-piperidin-2,6-dione), stereoisomer of lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione ), pomalidomide (4-amino-2-(2,6-dioxo-piperidin-3-yl)isoindol-1,3-dione) or avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazoline -3-yl)-piperidine-2,6-dione), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

[0048] In some embodiments, the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione, or a stereoisomer thereof, or their pharmaceutically acceptable salt, MES, hydrate, co-crystal, clathrate or polymorph. In some embodiments, the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione. In some embodiments, the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof , solvate, hydrate, co-crystal, clathrate or polymorph. In some embodiments, the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione.

[0049] In some embodiments of any of the methods provided herein, the immunomodulatory compound is administered orally, subcutaneously, or intravenously. In some embodiments, the immunomodulatory compound is administered orally. In some embodiments, the immunomodulatory compound is administered in a capsule or tablet.

[0050] In some embodiments of any of the methods provided herein, the immunomodulatory compound is administered in an amount of from or about 0.1 mg to about 100 mg, from or about 0.1 mg to 50 mg, from or about 0.1 mg to 25 mg, from or about 0.1 mg to 10 mg, from or about 0.1 mg to 5 mg, from or about 0.1 mg to 1 mg, from or about 1 mg up to 100 mg, from or about 1 mg to 50 mg, from or about 1 mg to 25 mg, from or about 1 mg to 10 mg, from or about 1 mg to 5 mg, from or about 5 mg up to 100 mg, from or about 5 mg to 50 mg, from or about 5 mg to 25 mg, from or about 5 mg to 10 mg, from or about 10 mg to 100 mg, from or about 10 mg up to 50 mg, from or from 10 mg to 25 mg, from or about 25 mg to 100 mg, from or about 25 mg to 50 mg, or from or about 50 mg to 100 mg, each inclusive.

[0051] In some embodiments of any of the methods provided herein, the immunomodulatory compound is administered once a day, twice a day, three times a day, four times a day, five times a day, or six times a day. In some embodiments, the immunomodulatory compound is administered at a total daily dosage amount of at least, or about, at least 0.1 mg per day, 0.5 mg per day, 1.0 mg per day, 2.5 mg per day, 5 mg per day, 10 mg per day, 25 mg per day, 50 mg per day, or 100 mg per day.

[0052] In some embodiments of any of the methods provided herein, the immunomodulatory compound is administered in an amount greater than or about greater than 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg, and less than 25 mg; or the immunomodulatory compound is administered in an amount greater than or about greater than 1 mg per day, 2.5 mg per day, 5 mg per day, 7.5 mg per day, 10 mg per day, 15 mg per day, and less than 25 mg per day day.

[0053] In some embodiments of any of the methods provided herein, the administration of a therapeutically effective amount of an immunomodulatory compound stimulates an increase in the proliferation of T-lymphocytes associated with T-cell therapy, compared with the reproduction after administration of T-cell therapy without an immunomodulatory compound.

[0054] In some embodiments of any of the methods provided herein, the administration of a therapeutically effective amount of an immunomodulatory compound stimulates an increase in T-lymphocyte-mediated cytolytic activity associated with T-cell therapy, compared with cytolytic activity after administration of T-lymphocytes without immunomodulatory connections.

[0055] In some embodiments of any of the methods provided herein, the administration of a therapeutically effective amount of an immunomodulatory compound stimulates an increase in cytokine production by T-lymphocytes associated with T-cell therapy, compared with the production of cytokines after administration of T-lymphocytes without an immunomodulatory compound . In some embodiments, this is greater than or about greater than 1.5x, 2.0x, 3.0x, 4.0x, 5.0x, 10.0x, or greater.

[0056] In some embodiments of any of the methods provided herein, the T cell therapy is or comprises tumor infiltrating lymphocyte (TIL) therapy or engineered cells expressing a recombinant receptor that specifically binds to an antigen. In some embodiments of any of the methods provided herein, the T cell therapy is or contains engineered cells expressing a recombinant receptor that specifically binds to an antigen. In some embodiments, the T cell therapy comprises cells expressing a recombinant receptor that is or contains a functional non-TCR antigen receptor, or a TCR or antigen-binding fragment thereof. In some embodiments, the recombinant antigen receptor is a chimeric antigen receptor (CAR).

[0057] In some embodiments of any of the methods provided herein, the T cell therapy comprises a recombinant antigen receptor that contains an extracellular domain containing an antigen-binding domain that specifically binds to an antigen. In some embodiments, the antigen is associated with, specific to, and/or expressed on a cell or tissue of the disease, disorder, or condition. In some embodiments, the disease, disorder, or condition is an infectious disease or disorder, an autoimmune disease, an inflammatory disease, or a tumor or cancer. In some embodiments, the antigen is a tumor antigen.

[0058] In some embodiments of any of the methods provided herein, the antigen is selected from ROR1, B-cell maturation antigen (BCMA), carbonic anhydrase 9 (CAIX), tEGFR, Her2/neu (erbB2 tyrosine kinase receptor), L1- CAM, CD19, CD20, CD22, mesothelin, CEA and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG -40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate-binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL- 22R-alpha, IL-13R-alpha2, kinase insert domain-containing receptor (kdr), kappa light chain, Ley antigen, L1 cell adhesion molecule, (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE -A3, MAGE-A6, preferably expressed melanoma antigen (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CA IX, HLA-AI MAGE Al, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, cancer-testicular antigen, mesothelin, mouse CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, G protein coupled receptor 5D (GPCR5D), oncofetal antigen , ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms tumor 1 (WT-1), cyclin, A2 cyclin, CCL-1, CD138, optionally, a human antigen to any of the above antigens; a pathogen-specific antigen; and an antigen associated with a universal label. In some embodiments, the antigen is or includes CD19, optionally human CD19. In some embodiments, the antigen is or includes an antigen associated with multiple myeloma, optionally BCMA, optionally human BCMA.

[0059] In some embodiments of any of the methods provided herein, the antigen binding domain is or contains an antibody or antibody fragment, which is optionally a single chain fragment. In some embodiments, the fragment contains the variable regions of the antibody connected by a flexible linker. In some embodiments, the fragment contains scFv.

[0060] In some embodiments of any of the methods provided herein, the T cell therapy comprises a recombinant receptor that further comprises a spacer, optionally derived from an immunoglobulin, optionally containing a hinge region. In some embodiments, the recombinant antigen receptor contains an intracellular signaling region. In some embodiments, the intracellular signaling region contains an intracellular signaling domain. In some embodiments, the intracellular signaling domain is or contains a primary signaling domain, a signaling domain that can induce a primary activation signal in a T-lymphocyte, a T-lymphocyte receptor (TCR) signaling domain component, and/or a signaling domain containing an immunoreceptor tyrosine activating motive (ITAM). In some embodiments of any of the methods provided herein, the intracellular signaling domain is or contains an intracellular signaling domain of the CD3 chain, optionally the CD3-zeta (CD3ζ) chain, or a signal portion thereof.

[0061] In some embodiments of any of the methods provided herein, the recombinant receptor further comprises a transmembrane domain located between the extracellular domain and the intracellular signaling region, where the transmembrane domain is optionally a CD8 or CD28 transmembrane domain. In some embodiments, the intracellular signaling region further includes a costimulatory signaling region. In some embodiments, the co-stimulatory signaling region comprises the intracellular signaling domain of the co-stimulatory T-lymphocyte molecule or a signaling portion thereof. In some embodiments, the co-stimulatory signaling region comprises a CD28, 4-1BB, or ICOS intracellular signaling domain, or a signaling portion thereof. In some embodiments, the co-stimulatory signaling region contains the 4-1BB intracellular signaling domain. In some embodiments of any of the methods provided herein, the costimulatory signaling region is located between the transmembrane domain and the intracellular signaling region.

[0062] In some embodiments of any of the methods provided herein, T cell therapy comprises:

T lymphocytes selected from central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central memory T lymphocytes, effector T lymphocytes and regulatory T lymphocytes; and/or a plurality of cells, the plurality comprising at least 50% of a population of cells selected from CD4+ T lymphocytes, CD8+ T lymphocytes, central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central Memory T-lymphocytes, effector T-lymphocytes and regulatory T-lymphocytes.

[0063] In some embodiments of any of the methods provided herein, the T cell therapy comprises T lymphocytes that are CD4+ or CD8+. In some embodiments, the T cell therapy comprises primary cells obtained from a subject. In some embodiments, the T cell therapy comprises cells that are autologous to the subject. In some embodiments, the T cell therapy comprises T lymphocytes that are allogeneic to the subject. In some embodiments of any of the methods provided herein, the subject is a human.

[0064] In some embodiments of any of the methods provided herein, T cell therapy comprises administering a total of, from or about 1 x 10 ⁵ to 1 x 10 ⁸ cells, T lymphocytes expressing recombinant receptors, in total , or peripheral blood mononuclear cells (PBMC), in total, from or about 5 × 10 ⁵ to 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in in total, or from or about 1 x 10 ⁶ to 1 x 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, each value inclusive.

[0065] In some embodiments of any of the methods provided herein, T cell therapy comprises the administration of no more than 1 x 10 ⁸ cells, in total, T lymphocytes expressing recombinant receptors in total, or peripheral blood mononuclear cells (PBMC ), in total, not more than 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in general, or peripheral blood mononuclear cells (PBMC), in total, not more than 0.5 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, no more than 1 × 10 ⁶ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or mononuclear cells peripheral blood cells (PBMC), in general, no more than 0.5 × 10 ⁶ cells, in general, T-lymphocytes expressing recombinant receptors, in general, or peripheral blood mononuclear cells (PBMC), in general.

[0066] In some embodiments of any of the methods provided herein, the number of cells administered in T cell therapy is less than that in another method in which T cell therapy is administered without administration of an immunomodulatory compound, optionally this the other method results in a similar or lower degree of alleviation or reduction or prevention of the disease or condition or symptom, or their severity, compared to the result of the present method. In some embodiments, the number of cells administered is 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, or 10-fold less than the amount administered in the other method.

[0067] In some embodiments of any of the methods provided herein, T cell therapy is administered as a separate pharmaceutical composition containing cells. In some embodiments of any of the methods provided herein, T cell therapy comprises a cell dose that is a divided dose, where the cells of the dose are administered in a plurality of compositions collectively containing all the cells of the dose, over a period of no more than three days.

[0068] In some embodiments of any of the methods provided herein, the method further comprises administering antilymphocyte chemotherapy prior to administering T cell therapy.

[0069] In some embodiments of any of the methods provided herein, the disease or condition is a cancer. In some embodiments, the cancer is B-lymphocyte cancer and/or myeloma, lymphoma, or leukemia. In some embodiments, the cancer is mantle cell lymphoma (MCL), multiple myeloma (MM), acute lymphocytic leukemia (ALL), adult ALL, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL), or diffuse large B cell lymphoma. (DLBCL). In some embodiments, the cancer is a non-hematologic cancer or is a solid tumor.

[0070] In some embodiments of any of the methods provided herein, T cell therapy demonstrates an increase or prolongation of proliferation and/or persistence in a subject compared to a method in which T cell therapy is administered to a subject without an immunomodulatory compound.

[0071] In some embodiments of any of the methods provided herein, the method reduces tumor mass to a greater extent and/or for a longer period of time compared to the reduction that would be observed for a comparable method in which T cell therapy is administered a subject without an immunomodulatory compound and/or in which an immunomodulatory compound is administered without T cell therapy, optionally at the same dose or dosing time schedule.

[0072] This document provides a kit containing a pharmaceutical composition containing a standard unit dose of T-cell therapy; and instructions for administering the composition to a subject having a disease or condition in conjunction with administering a composition comprising an immunomodulatory compound, wherein the instructions specify administering the immunomodulatory compound in one or more unit doses according to an administration cycle, including administering the immunomodulatory compound for up to 21 days, sequentially where the cycle includes more than 30 days from the start of the administration of the immunomodulatory compound; and/or administering an immunomodulatory compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or administration of an immunomodulatory compound for up to 14 consecutive days.

[0073] Also provided herein is a kit containing a pharmaceutical composition comprising one or more standard unit doses of an immunomodulatory compound; and instructions for administering an immunomodulatory compound to a subject having a disease or condition in conjunction with administering a unit dose of a pharmaceutical composition comprising T cell therapy, wherein the instructions specify administering one or more unit dose units of the immunomodulatory compound in accordance with an administration cycle including administration of the immunomodulatory compounds for up to 21 days, consecutively, where the cycle includes more than 30 days from the start of administration of the immunomodulatory compound; and/or administering an immunomodulatory compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or administration of an immunomodulatory compound for up to 14 consecutive days.

[0074] In some such embodiments, the instructions specify the initiation of administration of one or more standard unit doses of the immunomodulatory compound on the same day, optionally simultaneously, as the initiation of administration of T cell therapy. In some such embodiments, the instructions specify the initiation of administration of one or more standard unit doses of the immunomodulatory compound prior to the initiation of administration of T cell therapy.

[0075] In some such embodiments, the instructions specify commencing administration of one or more standard unit doses of the immunomodulatory compound one week or within a week prior to collection of a sample containing T lymphocytes to be genetically engineered from a subject, optionally when the sample is sample from apheresis; and/or during one or more steps of an ex vivo preparation method for receiving therapy with genetically engineered T-lymphocytes; and/or within 14 days prior to the introduction of T-cell therapy.

[0076] In some embodiments, one or more steps of the ex vivo preparation method is selected from isolating cells from a biological sample using leukapheresis or apheresis; selecting or enriching cells using immunoaffinity methods; introducing a recombinant nucleic acid, optionally a viral vector, into cells ; incubating cells, optionally genetically engineered cells, in the presence of one or more stimulation conditions; preparation of cells in the presence of a cryoprotectant; and/or preparing cells for administration to a subject, optionally in the presence of a pharmaceutically acceptable excipient.

[0077] In some such embodiments, the instructions specify the initiation of administration of one or more standard unit doses of the immunomodulatory compound within 10 days, 7 days, 4 days, 3 days, or 2 days prior to the administration of T cell therapy. In some examples, the instructions specify the initiation of administration of one or more standard unit doses of the immunomodulatory compound following the initiation of administration of T cell therapy. In some aspects, the instructions specify the start of administration of one or more standard unit doses of the immunomodulatory compound after at least 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least at least 4 weeks after the start of the administration of T-cell therapy, and/or 2-28 days or 7-21 days after the start of the administration of T-cell therapy.

[0078] Also provided herein is a kit containing a pharmaceutical composition comprising a standard unit dose of T cell therapy; and instructions for administering the composition to a subject having a disease or condition in conjunction with administering an immunomodulatory compound, wherein the instructions specify the start of administration of the immunomodulatory compound at one or more unit doses at a time point of at least 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the start of T-cell therapy administration, and/or it is carried out after 2-28 or after 7-21 days after the start of the introduction of T-cell therapy; and/or at or after a time point, optionally immediately after or within 1-3 days after: (i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0079] Provided herein is a kit containing a pharmaceutical composition comprising one or more standard unit doses of an immunomodulatory compound; and instructions for administering an immunomodulatory compound to a subject having a disease or condition in conjunction with administering a unit dose of a pharmaceutical composition comprising a T cell therapy, wherein the instructions specify the start of administration of one or more unit unit doses of the immunomodulatory compound at a time point of at least after 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the start of the introduction of T-cell therapy, and / or it is carried out through 2-28 or 7-21 days after the start of the introduction of T-cell therapy; and/or at or after a time point, optionally immediately after or within 1-3 days after: (i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0080] In some such embodiments, the instructions specify the initiation of administration of one or more standard unit doses of the immunomodulatory compound after a period of time that is greater than or about greater than 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 24 days, or 28 days after the start of T-cell therapy. In some such embodiments, the instructions specify the selection of a subject to administer one or more standard unit doses of an immunomodulatory compound, following administration of a T cell therapy, wherein: (i) a peak peak cell level from the T cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0081] This document provides a kit containing a pharmaceutical composition containing a standard unit dose of T-cell therapy; and instructions for administering the composition to a subject having a disease or condition in conjunction with administering an immunomodulatory compound, wherein the instructions specify administering the immunomodulatory compound to a subject in one or more unit doses if on or about day 12-15, optionally on the day On or about the 14th day after initiation of T-cell therapy for the treatment of the disease or condition, the cell count from T-cell therapy in the subject is less than 75% of the mean cell count from T-cell therapy at the same time in the plurality of subjects administered the same or similar dose of T-cell therapy; and/or the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, in blood less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl.

[0082] Provided herein is a kit containing a pharmaceutical composition comprising one or more standard unit doses of an immunomodulatory compound; and instructions for administering one or more standard unit doses of an immunomodulatory compound to a subject having a disease or condition in conjunction with administering a pharmaceutical composition comprising a unit dose of T cell therapy, wherein the instructions specify administering one or more unit doses of an immunomodulatory compound to a subject if on or about day 12-15, optionally on or about day 14, after initiation of T-cell therapy for the treatment of a disease or condition, the subject's T-cell therapy cell count is less than 75% of the mean cells from T-cell therapy at the same time in multiple subjects who are administered the same or similar dose of T-cell therapy; and/or the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, in blood less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl.

[0083] Provided herein are kits that contain (a) a pharmaceutical composition containing a unit dose of T cell therapy; and (b) instructions for administering the composition to a subject having a disease or condition in conjunction with administering a composition comprising an immunomodulatory compound, wherein said immunomodulatory compound is selected from the group consisting of: lenalidomide (3-(4-amino-1-oxo-1 ,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-diox-piperidin-3-yl)isoindol-1,3-dione) or avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomer, enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate thereof , hydrate, co-crystal, clathrate or polymorph, and where the instructions specify the administration of the immunomodulatory compound in one or more standard unit doses according to an administration cycle comprising: (i) administration of the immunomodulatory compound for up to 21 days, sequentially, where the cycle includes more than 30 days from starting the administration of the immunomodulatory compound; and/or (ii) administering an immunomodulatory compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or (iii) administering an immunomodulatory compound for up to 14 consecutive days.

[0084] Provided herein are kits that contain (a) a pharmaceutical composition containing one or more standard unit doses of an immunomodulatory compound; and (b) instructions for administering an immunomodulatory compound to a subject having a disease or condition, in conjunction with administering a standard unit dose of a pharmaceutical composition containing T cell therapy, wherein said immunomodulatory compound is selected from the group consisting of: lenalidomide (3-(4- amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol- 1,3-dione) or avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomer, enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, and wherein the instructions specify the administration of one or more standard unit doses of the immunomodulatory compound according to an administration cycle comprising: (i) administering the immunomodulatory compound for up to 21 days, sequentially, where the cycle includes more than 30 days from the start of the introduction of an immunomodulatory compound opinions; and/or (ii) administering an immunomodulatory compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or (iii) administering an immunomodulatory compound for up to 14 consecutive days.

[0085] Provided herein are kits that contain (a) a pharmaceutical composition containing a unit dose of T cell therapy; and (b) instructions for administering the composition to a subject having a disease or condition in conjunction with administering an immunomodulatory compound, wherein said immunomodulatory compound is selected from the group consisting of: lenalidomide (3-(4-amino-1-oxo-1,3- dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione) or avadomide (3 -(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomer, enantiomer or mixture of enantiomers, or their pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph, and where the instructions specify the start of administration of the immunomodulatory compound in one or more standard unit doses at the time point: (1) at least 2 days later, at least 1 week later, at least 2 weeks, at least 3 weeks or at least 4 weeks after the start of the introduction of T-cell therapy, and / or it is carried out after 2-2 8 or 7-21 days after the start of the introduction of T-cell therapy; and/or (2) at or after a time point, optionally immediately thereafter or within 1-3 days after: (i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0086] Provided herein are kits that comprise (a) a pharmaceutical composition comprising one or more standard unit doses of an immunomodulatory compound, wherein said immunomodulatory compound is selected from the group consisting of: lenalidomide (3-(4-amino-1-oxo -1,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione ) or avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), a stereoisomer, enantiomer or mixture of enantiomers thereof, or a pharmaceutically acceptable salt thereof , solvate, hydrate, co-crystal, clathrate or polymorph; and (b) instructions for administering an immunomodulatory compound to a subject having a disease or condition in conjunction with administering a unit dose of a pharmaceutical composition comprising a T cell therapy, wherein the instructions specify the start of administration of one or more unit unit doses of the immunomodulatory compound at a time point: ( 1) at least 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the start of T-cell therapy, and/or it is carried out after 2-28 or 7-21 days after the start of the introduction of T-cell therapy; and/or (2) at or after a time point, optionally immediately thereafter or within 1-3 days after: (i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0087] Provided herein are kits that contain (a) a pharmaceutical composition containing a unit dose of T cell therapy; and (b) instructions for administering the composition to a subject having a disease or condition in conjunction with administering an immunomodulatory compound, wherein said immunomodulatory compound is selected from the group consisting of: lenalidomide (3-(4-amino-1-oxo-1,3- dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione) or avadomide (3 -(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomer, enantiomer or mixture of enantiomers, or their pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph, and wherein the instructions specify administering the immunomodulatory compound to a subject in one or more unit doses if on or about day 12-15, optionally on or about day 14, after initiation of T-cell administration. therapy for the treatment of a disease or condition: (i) the number of cells from T-cell therapy in the subject is less than 75% of the average number cells from T-cell therapy at the same time in a plurality of subjects receiving the same or similar dose of T-cell therapy; and/or (ii) the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, in the blood is less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl.

[0088] Provided herein are kits that comprise (a) a pharmaceutical composition comprising one or more standard unit doses of an immunomodulatory compound, wherein said immunomodulatory compound is selected from the group consisting of: lenalidomide (3-(4-amino-1-oxo -1,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione ) or avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomer, enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt thereof , solvate, hydrate, co-crystal, clathrate or polymorph; and (b) instructions for administering one or more unit dose units of an immunomodulatory compound to a subject having a disease or condition in conjunction with administering a pharmaceutical composition comprising a unit dose of T cell therapy, wherein the instructions specify the administration of one or more unit unit doses of the immunomodulatory compound to a subject, if on or about day 12-15, optionally, on or about day 14, after initiation of T-cell therapy administration for the treatment of a disease or condition: (i) the number of cells from T-cell therapy in the subject less than 75% of the average number of cells from T-cell therapy at the same time in multiple subjects who are administered the same or similar dose of T-cell therapy; and/or (ii) the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, in the blood is less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl.

[0089] In some such embodiments, the immunomodulatory compound is formulated in an amount for daily administration and/or instructions specify daily administration of the immunomodulatory compound. In some such embodiments, the instructions specify administering the immunomodulatory compound for more than or about more than 7 consecutive days, more than or about more than 14 consecutive days, more than or about more than 21 consecutive days, more than or about more than 21 consecutive days, or more than or about more 28 consecutive days.

[0090] In some such embodiments, the instructions specify the administration of the immunomodulatory compound in an administration cycle , including daily administration for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered. In some examples, the instructions specify a rest period during which no immunomodulatory compound is administered, greater than 7 consecutive days, greater than 14 consecutive days, greater than 21 days, or greater than 28 days. In some such embodiments, the instructions specify an immunomodulatory compound administration cycle that is repeated at least once.

[0091] In some such embodiments, the instructions specify the continuation of the administration of the immunomodulatory compound, at least from the time after the start of the administration of T-lymphocytes, until the number of cells from the administered T-cell therapy, or derived from them, is detected in the subject's blood will not increase compared to the subject's cell count at the time immediately prior to the administration of the immunomodulatory compound or compared to the time prior to the administration of the T-cell therapy; until the number of cells from or derived from T-cell therapy detected in the blood is within 2.0 times (more or less) the peak or maximum number observed in the blood of the subject after the start of T-lymphocyte administration; until the number of cells from T-cell therapy detected in the subject's blood is greater than or about greater than 10%, 15%, 20%, 30%, 40%, 50%, or 60% of the total number of peripheral blood mononuclear cells (PBMC) in subject's blood; and/or until the subject demonstrates a reduction in tumor weight compared to tumor weight at the time immediately prior to administration of the T cell therapy or at the time immediately prior to administration of the immunomodulatory compound; and/or until the subject demonstrates complete or clinical remission.

[0092] In some such embodiments, the immunomodulatory compound binds to Cereblon (CRBN) and/or to the CRBN-E3 ubiquitin ligase complex; and/or inhibits the growth transcription factor Ikaros (IKZF1) or Aiolos (IKZF3); and/or enhances ubiquitination or degradation of Ikaros (IKZF1) or Aiolos (IKZF3). In some such embodiments, the immunomodulatory compound is thalidomide or is a derivative or analogue of thalidomide. In some such embodiments, the immunomodulatory compound is lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione) or pomalidomide (4- amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione), avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)- piperidin-2,6-dione), stereoisomer of lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4- amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione), avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)- piperidine-2,6-dione) or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In some such embodiments, the immunomodulatory compound is lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione), a stereoisomer of lenalidomide (3 -(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione) or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

[0093] In some such embodiments, the immunomodulatory compound is formulated for oral, subcutaneous, or intravenous administration. In some examples, the immunomodulatory compound is formulated for oral administration. In some such embodiments, the immunomodulatory compound is formulated into capsules or tablets.

[0094] In some such embodiments, each of the one or more unit dose unit doses of the immunomodulatory compound contains an amount of from or about 0.1 mg to about 100 mg, from or about 0.1 mg to 50 mg, from or about 0.1 mg to 25 mg, from or about 0.1 mg to 10 mg, from or about 0.1 mg to 5 mg, from or about 0.1 mg to 1 mg, from or about 1 mg up to 100 mg, from or about 1 mg to 50 mg, from or about 1 mg to 25 mg, from or about 1 mg to 10 mg, from or about 1 mg to 5 mg, from or about 5 mg up to 100 mg, from or about 5 mg to 50 mg, from or about 5 mg to 25 mg, from or about 5 mg to 10 mg, from or about 10 mg to 100 mg, from or about 10 mg up to 50 mg, from or from 10 mg to 25 mg, from or about 25 mg to 100 mg, from or about 25 mg to 50 mg, or from or about 50 mg to 100 mg, each inclusive; and/or each of one or more standard unit doses of the immunomodulatory compound contains an amount of at least, or about, at least 0.1 mg, 0.5 mg, 1.0 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg or 100 mg. In some such embodiments, one or more unit doses of the immunomodulatory compound contains an amount greater than or about greater than 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg, and less than 25 mg.

[0095] In some such embodiments, the T cell therapy is or comprises tumor infiltrating lymphocyte (TIL) therapy or engineered cells expressing a recombinant receptor that specifically binds to an antigen. In some such embodiments, the T cell therapy is or contains engineered cells expressing a recombinant receptor that specifically binds to an antigen.

[0096] In some such embodiments, the recombinant receptor is or contains a functional non-TCR antigen receptor, or a TCR or antigen-binding fragment thereof. In some such embodiments, the recombinant antigen receptor is a chimeric antigen receptor (CAR). In some such embodiments, the recombinant antigen receptor comprises an extracellular domain containing an antigen-binding domain that specifically binds to an antigen.

[0097] In some such embodiments, the antigen is associated with, specific to, and/or expressed on a cell or tissue of the disease, disorder, or condition. In some instances, the disease, disorder, or condition is an infectious disease or disorder, an autoimmune disease, an inflammatory disease, or a tumor or cancer.

[0098] In some such embodiments, the antigen is a tumor antigen. In some such embodiments, the antigen is selected from ROR1, B cell maturation antigen (BCMA), charcoal anhydrase 9 (CAIX), tEGFR, Her2/neu (erbB2 tyrosine kinase receptor), L1-CAM, CD19, CD20, CD22, mesothelin, CEA and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain-containing receptor (kdr), kappa light chain, Ley antigen, L1 cell adhesion molecule, (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, preferably expressed antigen melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE Al, HLA-A2 NY- ESO-1, PSCA, receptor-a folates, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, testicular cancer antigen, mesothelin, mouse CMV, mucin 1 (MUC1), MUC16, PSCA , NKG2D, NY-ESO-1, MART-1, gp100, G protein coupled receptor 5D (GPCR5D), oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms tumor 1 (WT-1), cyclin, cyclin A2, CCL-1, CD138, optional, human antigen to any object from the above; a pathogen-specific antigen; and an antigen associated with a universal label. In some embodiments, the antigen is or includes CD19, optionally human CD19. In some embodiments, the antigen is or includes BCMA, optionally human BCMA.

[0099] In some such embodiments, the antigen-binding fragment is or contains an antibody or antibody fragment, which is optionally a single chain fragment. In some cases, the fragment contains the variable regions of the antibody connected by a flexible linker. In some embodiments, the fragment comprises a single chain scFv variable fragment. In some such embodiments, the recombinant receptor further comprises a spacer, optionally derived from an immunoglobulin, optionally containing a hinge region.

[0100] In some such embodiments, the recombinant antigen receptor contains an intracellular signaling region. In some such embodiments, the intracellular signaling region contains an intracellular signaling domain. In some examples, the intracellular signaling domain is or contains a primary signaling domain, a signaling domain that can induce a primary activation signal in a T-lymphocyte, a T-lymphocyte receptor (TCR) signaling domain component, and/or a signaling domain containing an immunoreceptor tyrosine activating motif. (ITAM).

[0101] In some such embodiments, the intracellular signaling domain is or comprises an intracellular signaling domain of the CD3 chain, optionally the CD3-zeta (CD3ζ) chain, or a signal portion thereof. In some such embodiments, the recombinant receptor further comprises a transmembrane domain located between the extracellular domain and the intracellular signaling region, where the transmembrane domain is optionally a CD8 or CD28 transmembrane domain.

[0102] In some such embodiments, the intracellular signaling region further comprises a costimulatory signaling region. In some cases, the costimulatory signaling region contains the intracellular signaling domain of the costimulatory T-lymphocyte molecule or a signaling portion thereof. In some embodiments, the co-stimulatory signaling region comprises a CD28, 4-1BB, or ICOS intracellular signaling domain, or a signaling portion thereof. In some examples, the costimulatory signaling region contains the 4-1BB intracellular signaling domain. In some such embodiments, the costimulatory signaling region is located between the transmembrane domain and the intracellular signaling region.

[0103] In some such embodiments, the T cell therapy comprises T lymphocytes selected from the group consisting of central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central memory T lymphocytes, effector T -lymphocytes and regulatory T-lymphocytes; and/or a plurality of cells, the plurality being at least 50% of a population of cells selected from the group consisting of CD4+ T lymphocytes, CD8+ T lymphocytes, central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes , stem central memory T-lymphocytes, effector T-lymphocytes and regulatory T-lymphocytes. In some such embodiments, the T cell therapy comprises T lymphocytes that are CD4+ or CD8+.

[0104] In some such embodiments, the T cell therapy comprises primary cells obtained from a subject. In some such embodiments, the T cell therapy is autologous to the subject. In some such embodiments, the T cell is allogeneic to the subject. In some such embodiments, the subject is a human.

[0105] In some such embodiments, a unit dose of T-cell therapy contains from or about 1 x 10 ⁵ to 1 x 10 ⁸ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, from or about 5 × 10 ⁵ to 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, or from or from about 1 x 10 ⁶ to 1 x 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, each value is inclusive. In some such embodiments, a standard unit dose of T cell therapy comprises administering no more than 1 x 10 ⁸ cells, in total, no more than total recombinant receptor-expressing T lymphocytes, or no more than total peripheral blood mononuclear cells (PBMCs). than 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in general, or peripheral blood mononuclear cells (PBMC), in total, not more than 0.5 × 10 ⁷ cells, in total, T-lymphocytes, expressing recombinant receptors in total, or peripheral blood mononuclear cells (PBMC) in total, not more than 1 × 10 ⁶ cells in total, T-lymphocytes expressing recombinant receptors in total, or peripheral blood mononuclear cells (PBMC) in total in general, not more than 0.5×10 ⁶ cells in total, T-lymphocytes expressing recombinant receptors in general, or peripheral blood mononuclear cells (PBMCs) in general.

[0106] In some such embodiments, a unit dose of T cell therapy comprises a cell dose that is a divided dose, where the cells of the dose are administered in a plurality of compositions collectively containing all the cells of the dose, over a period of no more than three days.

[0107] In some such embodiments, the instructions further specify the administration of anti-lymphocyte chemotherapy prior to the administration of T cell therapy. In some such embodiments, the disease or condition is a cancer. In some such embodiments, the cancer is B-lymphocyte cancer and/or myeloma, lymphoma, or leukemia. In some embodiments, the cancer is mantle cell lymphoma (MCL), multiple myeloma (MM), acute lymphocytic leukemia (ALL), adult ALL, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL), or diffuse large B cell lymphoma ( DLBCL). In some cases, the cancer is a non-hematologic cancer or is a solid tumor.

[0108] This document proposes an industrial product containing any of the kits described in this document.

[0109] Also provided herein is a pharmaceutical composition comprising a T cell therapy, an immunomodulatory compound, and a pharmaceutically acceptable carrier. In some embodiments, the T cell therapy is formulated in a unit dose amount. In some cases, a standard unit dose of T-cell therapy contains from or about 1 x 10 ⁵ to 1 x 10 ⁸ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in in total, from or about 5 × 10 ⁵ to 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, or from or from about 1 × 10 ⁶ to 1 x 10 ⁷ cells in total, T-lymphocytes expressing recombinant receptors in total, or peripheral blood mononuclear cells (PBMC) in total, each value inclusive.

[0110] Provided herein are pharmaceutical compositions that include T cell therapy, an immunomodulatory compound, and a pharmaceutically acceptable carrier, wherein said immunomodulatory compound is selected from the group consisting of: lenalidomide (3-(4-amino-1-oxo-1, 3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4-amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione) or avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione), their stereoisomer, enantiomer or mixture of enantiomers, or their pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph.

[0111] In some embodiments, a standard unit dose of T cell therapy comprises administering no more than 1 x 10 ⁸ cells, in total, T lymphocytes expressing recombinant receptors in total, or peripheral blood mononuclear cells (PBMCs), in total, not more than 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in general, or peripheral blood mononuclear cells (PBMC), in total, not more than 0.5 × 10 ⁷ cells, in total, T- recombinant receptor-expressing lymphocytes in total or peripheral blood mononuclear cells (PBMC) in total not more than 1 × 10 ⁶ cells in total recombinant receptor-expressing T-lymphocytes in total or peripheral blood mononuclear cells (PBMC) , in general, not more than 0.5 × 10 ⁶ cells, in general, T-lymphocytes expressing recombinant receptors, in general, or peripheral blood mononuclear cells (PBMC), in general.

[0112] In some such embodiments, the immunomodulatory compound binds to Cereblon (CRBN) and/or to the CRBN and E3 ubiquitin ligase complex; and/or inhibits the growth transcription factor Ikaros (IKZF1) or Aiolos (IKZF3); and/or enhances ubiquitination or degradation of Ikaros (IKZF1) or Aiolos (IKZF3). In some such embodiments, the immunomodulatory compound is thalidomide or is a derivative or analogue of thalidomide. In some such embodiments, the immunomodulatory compound is lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione) or pomalidomide (4- amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione), avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)- piperidin-2,6-dione), stereoisomer of lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione), pomalidomide (4- amino-2-(2,6-dioxopiperidin-3-yl)isoindol-1,3-dione), avadomide (3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)- piperidine-2,6-dione) or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In some such embodiments, the immunomodulatory compound is lenalidomide (3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione), a stereoisomer of lenalidomide (3 -(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione) or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

[0113] In some such embodiments, the immunomodulatory compound is formulated in a unit dose amount. In some such embodiments, the amount of the immunomodulatory compound in the composition is from or about 0.1 mg to about 100 mg, from or about 0.1 mg to 50 mg, from or about 0.1 mg to 25 mg, from or about 0.1 mg to 10 mg, from or about 0.1 mg to 5 mg, from or about 0.1 mg to 1 mg, from or about 1 mg to 100 mg, from or about 1 mg to 50 mg, from or about 1 mg to 25 mg, from or about 1 mg to 10 mg, from or about 1 mg to 5 mg, from or about 5 mg to 100 mg, from or about 5 mg to 50 mg, from or about 5 mg to 25 mg, from or about 5 mg to 10 mg, from or about 10 mg to 100 mg, from or about 10 mg to 50 mg, from or from 10 mg up to 25 mg, from or about 25 mg to 100 mg, from or about 25 mg to 50 mg, or from or about 50 mg to 100 mg, each inclusive; and/or the amount of the immunomodulatory compound in the composition is at least or approximately at least 0.1 mg, 0.5 mg, 1.0 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg or 100 mg. In some such embodiments, the amount of the immunomodulatory compound in the composition is greater than or greater than about 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg, and less than 25 mg.

[0114] In some such embodiments, the T cell therapy is or comprises tumor infiltrating lymphocyte (TIL) therapy or engineered cells expressing a recombinant receptor that specifically binds to an antigen. In some such embodiments, the T cell therapy is or contains engineered cells expressing a recombinant receptor that specifically binds to an antigen. In some such embodiments, the recombinant receptor is or contains a functional non-TCR antigen receptor, or a TCR or antigen-binding fragment thereof. In some such embodiments, the recombinant antigen receptor is a chimeric antigen receptor (CAR).

[0115] In some such embodiments, the recombinant antigen receptor comprises an extracellular domain containing an antigen-binding domain that specifically binds to an antigen. In some such embodiments, the antigen is associated with, specific to, and/or expressed on a cell or tissue of the disease, disorder, or condition. In some instances, the disease, disorder, or condition is an infectious disease or disorder, an autoimmune disease, an inflammatory disease or tumor, or a cancer.

[0116] In some such embodiments, the antigen is a tumor antigen. In some such embodiments, the antigen is selected from ROR1, B cell maturation antigen (BCMA), charcoal anhydrase 9 (CAIX), tEGFR, Her2/neu (erbB2 tyrosine kinase receptor), L1-CAM, CD19, CD20, CD22, mesothelin, CEA and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain-containing receptor (kdr), kappa light chain, Ley antigen, L1 cell adhesion molecule, (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, preferably expressed antigen melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE Al, HLA-A2 NY- ESO-1, PSCA, Pho receptor-a lats, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, fetal AchR, NKG2D ligands, CD44v6, dual antigen, testicular cancer antigen, mesothelin, mouse CMV, mucin 1 (MUC1), MUC16, PSCA , NKG2D, NY-ESO-1, MART-1, gp100, G protein coupled receptor 5D (GPCR5D), oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms tumor 1 (WT-1), cyclin, cyclin A2, CCL-1, CD138, optional, human antigen to any of the above; a pathogen-specific antigen; and an antigen associated with a universal label. In some such embodiments, the antigen is or comprises CD19, optionally human CD19. In some embodiments, the antigen is or contains BCMA, optionally human BCMA.

[0117] In some such embodiments, the antigen-binding fragment is or contains an antibody or antibody fragment, which is optionally a single chain fragment. In some examples, the fragment contains the variable regions of the antibody connected by a flexible linker. In some embodiments, the fragment contains scFv. In some such embodiments, the recombinant receptor further comprises a spacer, optionally derived from an immunoglobulin, optionally containing a hinge region.

[0118] In some such embodiments, the recombinant antigen receptor contains an intracellular signaling region. In some examples, the intracellular signaling region contains an intracellular signaling domain. In some cases, the intracellular signaling domain is or contains a primary signaling domain, a signaling domain that can induce a primary activation signal in a T-lymphocyte, a T-lymphocyte receptor (TCR) signaling domain component, and/or a signaling domain containing an immunoreceptor tyrosine activating motif. (ITAM). In some embodiments, the intracellular signaling domain is or contains an intracellular signaling domain of the CD3 chain, optionally the CD3-zeta (CD3ζ) chain, or a signal portion thereof.

[0119] In some such embodiments, the recombinant receptor further comprises a transmembrane domain located between the extracellular domain and the intracellular signaling region, where the transmembrane domain is optionally a CD8, CD28, CTLA-4, or PD-1 transmembrane domain. In some such embodiments, the intracellular signaling region further comprises a costimulatory signaling region.

[0120] In some such embodiments, the co-stimulatory signaling region comprises an intracellular signaling domain of a co-stimulatory T-lymphocyte molecule or a signaling portion thereof. In some embodiments, the co-stimulatory signaling region comprises a CD28, 4-1BB, or ICOS intracellular signaling domain, or a signaling portion thereof. In some examples, the costimulatory signaling region contains the 4-1BB intracellular signaling domain. In some such embodiments, the costimulatory signaling region is located between the transmembrane domain and the intracellular signaling region. In some such embodiments, the recombinant receptor is or comprises a chimeric antigen receptor comprising an antigen binding domain, a spacer, a transmembrane domain from CD28, an intracellular signaling domain containing a CD3-zeta chain (CD3ζ), and an intracellular signaling domain from 4-1BB .

[0121] In some such embodiments, the T cell therapy comprises T lymphocytes selected from the group consisting of central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central memory T lymphocytes, effector T -lymphocytes and regulatory T-lymphocytes; and/or a plurality of cells, the plurality being at least 50% of a population of cells selected from the group consisting of CD4+ T lymphocytes, CD8+ T lymphocytes, central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central memory T-lymphocytes, effector T-lymphocytes and regulatory T-lymphocytes. In some such embodiments, the T cell therapy comprises T lymphocytes that are CD4+ or CD8+. In some examples, the ratio of CD4+ T lymphocytes to CD8+ T lymphocytes is from or about 1:3 to 3:1, optionally 1:1.

[0122] In some such embodiments, the T cell therapy comprises primary cells obtained from a subject. In some cases, the subject is a human.

[0123] In some such embodiments, the pharmaceutical composition comprises a volume of from or about 1 ml to 100 ml, 1 ml to 75 ml, 1 ml to 50 ml, 1 ml to 25 ml, 1 ml to 10 ml , 1 ml to 5 washes 5 ml to 100 ml, 5 ml to 75 ml, 5 ml to 50 ml, 5 ml to 25 washes 5 ml to 10 ml, 10 ml to 100 ml, 10 ml to 75 ml, 10 ml to 50 ml, 10 ml to 25 ml, 25 ml to 100 ml, 25 ml to 75 ml, 25 ml to 50 ml, 50 ml to 100 ml, 50 ml to 75 ml or from 75 ml to 100 ml. In some such embodiments, the pharmaceutical composition contains a volume of at least, or about, at least, or about 1 ml, 5 ml, 10 ml, 20 ml, 25 ml, 30 ml, 40 ml, 50 ml, 60 ml , 70 ml, 80 ml, 90 ml or 100 ml.

[0124] In some such embodiments, the pharmaceutical composition further comprises a cryoprotectant. In some such embodiments, the pharmaceutical composition is sterile.

[0125] This document proposes an industrial product containing any of the pharmaceutical compositions described in this document.

[0126] A method of treatment is also provided, comprising administering any of the pharmaceutical compositions described herein to a subject for the treatment of a disease or condition. In some cases, the disease or condition is a cancer. In some cases, the cancer is B-lymphocyte cancer and/or myeloma, lymphoma, or leukemia. In some embodiments, the cancer is mantle cell lymphoma (MCL), multiple myeloma (MM), acute lymphocytic leukemia (ALL), adult ALL, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL), diffuse large B cell lymphoma ( DLB) or follicular lymphoma (FL).

Brief description of the drawings

[0127] Figure 1A shows surface expression of BCMA by multiple myeloma cell lines (RPMI-8226, MM1.S and OPM-2). The dashed line shows the background of the assay and BCMA-negative cell lines are stained with anti-BCMA antibody. MFI - median fluorescence intensity.

[0128] Figure 1B shows the percent reduction in BCMA-expressing target cells (RPMI-8226) by anti-BCMA CAR+ T lymphocytes in the presence and absence of lenalidomide (10 μM) after 6 days of co-culture. 1C shows the effect of lenalidomide on the cytolytic activity of anti-BCMA CAR+ T lymphocytes against RPMI-8226 target cells.

[0129] Figures 2A-2C show the amount of IL-2 (Figure 2A), IFNγ (Figure 2B) and TNF-α (Figure 2C) observed in supernatants after incubation of target RPMI-8226 cells with CAR anti -BCMA in the presence in the presence and in the absence.

[0130] Figure 3A shows the effect of increasing lenalidomide concentrations on the cytolytic activity of anti-BCMA CAR+ T lymphocytes against target OPM2 cells.

[0131] Figures 3B-D show the amount of IFNγ (FIG. 3B), IL-2 (FIG. 3C), and TNF-α (FIG. 3D) observed in supernatants after target OPM2 cells were incubated with anti-BCMA CAR T lymphocytes in the presence of elevated concentrations of lenalidomide, or in the absence of lenalidomide.

[0132] Figure 3E shows antigen-specific cytolytic activity of CAR-T anti-BCMA and cytokine production by CAR+ anti-BCMA T lymphocytes derived from representative healthy donors and multiple myeloma patients against target OPM-2 cells in the presence of various concentrations lenalidomide (0.01 μM, 0.1 μM, 1.0 μM or 10 μM lenalidomide) or in the absence of lenalidomide.

[0133] Figure 3F shows the antigen-specific cytolytic activity of CAR-T anti-BCMA T-lymphocytes CAR+ anti-BCMA derived from three healthy donors and one multiple myeloma patient against target OPM-2 and RPMI-8226 cells in the presence of various concentrations of lenalidomide (0.01 μM, 0.1 μM, 1.0 μM or 10 μM lenalidomide) or in the absence of lenalidomide. 3G shows cytokine production by anti-BCMA CAR+ T lymphocytes derived from three healthy donors and one multiple myeloma patient against target OPM-2 cells in the presence of various concentrations of lenalidomide (0.01 μM, 0.1 μM, 1, 0 µM or 10 µM lenalidomide) or in the absence of lenalidomide. 3H shows cytokine production by anti-BCMA CAR+ T lymphocytes derived from three healthy donors and one multiple myeloma patient against target RPMI-8226 cells in the presence of various concentrations of lenalidomide (0.01 μM, 0.1 μM, 1, 0 µM or 10 µM lenalidomide) or in the absence of lenalidomide.

[0134] Figure 4A shows the expansion of anti-BCMA CAR T lymphocytes after re-stimulation in the presence of various concentrations of lenalidomide.

[0135] Figure 4B shows the expansion of anti-BCMA CAR T lymphocytes after re-stimulation in both the presence and absence of lenalidomide.

[0136] Figure 5A shows cell counts (projective population doublings) of anti-BCMA CAR+ T lymphocytes from three donors at each time point in the restimulation assay, in the presence of vehicle or 0.1 μM lenalidomide. “×” indicates the insufficiency of cells for reinoculation in the assay. 5B shows CD25 median fluorescence intensity (MFI) (tuned to live CD3+ CAR+). 5C shows cytokine production normalized to the number of cells inoculated (upper and lower left panels) and CD25 median fluorescence intensity (MFI) (tuned to live CD3+ CAR+) (lower right panel).

[0137] Figure 6A shows the total number of CD3+ cells in culture on days 2 and 7 after anti-BCMA CAR T-lymphocyte incubation. or T-lymphocytes that do not express CAR (sham), in the presence or absence of lenalidomide. Figures 6B and 6C show CD25+ expression in CD4+ (FIG. 6B) and CD8+ (FIG. 6C) T lymphocytes in culture on days 2 and 7 after anti-BCMA CAR T lymphocyte incubation. or T lymphocytes that do not express CAR (mock) in the presence or absence of lenalidomide.

[0138] Figure 7A shows tumor volume in mice as a function of time after administration of a low dose of anti-BCMA CAR+ T-lymphocytes in the presence, presence and absence.

[0139] FIG. 7B shows the percentage survival of mice treated with a low dose of anti-BCMA CAR+ T-lymphocytes in the presence, presence and absence. For control groups, T-lymphocytes that do not express CAR (sham), in the presence in the presence and in the absence, are administered, and lenalidomide is also administered without T-lymphocytes.

[0140] Fig.8A shows the levels of T-lymphocytes CD4+ CAR+ in the blood of mice treated with CAR+ T lymphocytes anti-BCMA and lenalidomide compared with the other treatment groups on days 7 and 14. FIG. BCMA and lenalidomide compared with other treatment groups on days 21 and 36. Figure 8C shows blood levels of CD8+ CAR+ T lymphocytes in mice treated with anti-BCMA CAR+ T lymphocytes and lenalidomide compared with other treatment groups on days 7 and 14. FIG. 8D shows blood levels of CD8+ CAR+ T lymphocytes in mice treated with anti-BCMA CAR+ T lymphocytes and lenalidomide compared to other treatment groups on days 21 and 36.

[0141] FIG. 8E shows blood levels of CD4+ CAR+ T lymphocytes in mice treated with non-CAR+ T lymphocytes and lenalidomide compared with other treatment groups on days 7 and 14. FIG. 8F shows levels of CD4+ CAR+ T lymphocytes. in the blood of mice treated with non-CAR+ T lymphocytes and lenalidomide compared with other treatment groups on days 21 and 36. Figure 8G shows the blood levels of CD8+ CAR+ T lymphocytes in mice treated with non-CAR+ T lymphocytes and lenalidomide compared with other treatment groups on days 7 and 14. FIG. 8H shows blood levels of CD8+ CAR+ T lymphocytes in mice treated with non-CAR+ T lymphocytes and lenalidomide compared with other treatment groups on days 21 and 36.

[0142] Figures 9A and 9B show the results of tumor weight in mice treated with Regimen A (LenA), in which mice are treated with lenalidomide one day prior to administration of CAR+ T-lymphocytes.

[0143] Figure 9C shows single mouse tumor weight as a function of time to day 53.

[0144] FIG. 9D shows the results as images of the tumor at day 46 after CAR+ cell injection for a single mouse receiving a higher dose of CAR+ (1 x 10 ⁶ ) with lenalidomide at day -1 (Len A). Fig. 9E shows the results as images of the tumor at day 46 post CAR+ cell injection for a single mouse receiving a higher dose of CAR+ (1 x 10 ⁶ ) without lenalidomide on day -1 (Len A).

[0145] Figures 9F and 9G show the results on tumor weight of mice treated according to regimen B (LenB), in which administration of lenalidomide begins on day 14 after administration of CAR+ T.

[0146] Figure 9H shows tumor mass of an individual mouse over time to day 53.

[0147] Figure 9I shows the results as a tumor image (day 46 after CAR+ cell injection) for a single mouse receiving a higher dose of CAR+ (1 x 10 ⁶ ) with lenalidomide on day -1 (Len A). FIG. 9J shows the results as a tumor image (day 46 after CAR+ cell injection) for a single mouse receiving a higher dose of CAR+ (1 x 10 ⁶ ) without lenalidomide on day -1 (Len A).

[0148] Figures 10A-10D show the survival of mice in the presence or absence of lenalidomide. Lenalidomide is administered via Mode A (Len A; lenalidomide administration begins on day -1) or Mode B (Len B; lenalidomide administration begins on day 14) in combination with low (5 × 10 ⁵ or 5e ⁵ ) or high (1 × 10 ⁶ or 1e ⁶ ) doses of CAR+ T-lymphocytes. For control groups, T-lymphocytes that do not express CAR (sham) are administered in the presence in the presence and absence of both Mode A and Mode B, and lenalidomide without T-lymphocytes is also administered through both Mode A and Mode b.

[0149] Figure 10E shows the results of tumor weight assessments for mice receiving a higher dose of CAR+ (1 x 10 ⁶ ) and receiving daily intraperitoneal administration of 10 mg/kg lenalidomide or vehicle control, or starting on day -1 (the day before administration of CAR-T) (simultaneously with lenalidomide (lenalidomide (C) or vehicle (vehicle (C)) on day 14 after administration of CAR T-lymphocytes (or dummy cells) (delayed lenalidomide (D)). Results are shown up to day 60 as analyzed by bioluminescence measured by flow cytometry Figure 10F shows the percent survival of mice in the presence or absence of lenalidomide Figures 10G and 10H show flow cytometric analysis of control dummy cells and CAR T-lymphocytes in the blood of mice at day 8, 14, 22 and 28 after injection of CAR T-lymphocytes from two donors.

[0150] Figure 11 shows the number of CD4+ and CD8+ T lymphocytes in cultures of anti-CD19 CAR T lymphocytes stimulated with a suboptimal concentration of anti-CD3 in the presence, presence and absence of lenalidomide.

[0151] FIG. 12A shows peripheral blood CD3+/CAR+ T-lymphocyte counts measured at specific time points post-infusion for subjects grouped by best overall response.

[0152] FIG. 12B shows peripheral blood CD3+/CAR+ T-lymphocyte counts measured at specific time points after infusion in subjects that achieve a response, grouped by continuous response at 3 months.

[0153] Figures 12C-12D show CD4+/CAR ⁺ and CD8+/CAR+ T-lymphocyte levels in peripheral blood measured at specific time points after infusion in subjects that achieve a response, grouped by continuous response at 3 months.

[0154] FIG. 13A shows the number of CD3+/CAR+, CD4+/CAR+, CD8+/CAR+ T lymphocytes in the peripheral blood of a subject with chemotherapy-resistant DLBCL, measured at specific time points. 13B shows a pre-treatment axial PET-CT image showing intracranial abnormality in the right middle cranial fossa and extensive subcutaneous tissue abnormality in the right posterior auricular region. Fig. 13C is a post-treatment PET-CT image showing resolution of the anomaly in Fig. 13B after treatment with anti-CD19 CAR+ T lymphocytes. 13D is a pre-treatment MRI image of the brain (T ₁ -weighted high resolution image using contrast material; axial view) showing a uniform increase in mass in the right middle cranial fossa. 13E is a post-treatment MRI image showing almost complete resolution of the mass increase. 13F is an axial PET-CT image at relapse showing the reappearance of a right posterior auricular tumor associated with heavy consumption ^{of 18} F-fluorodeoxyglycose (arrow). 13G is a PET-CT image showing resolution of a posterior auricular tumor after incisional biopsy and reexpansion of CAR+ T-lymphocytes.

[0155] Figure 14 shows the level of viable target cells over a period of approximately 120 hours when anti-CD19 CAR+ T lymphocytes are incubated with K562-CD19 effector cells at an effector to target cell ratio (E:T) of 5:1 in the presence of or in the absence of 1 nM, 5 nM, 60 nM, 550 nM or 5000 nM lenalidomide or in the absence of lenalidomide (control).

[0156] Figure 15A shows levels of CD25+ expression in both CD4+ and CD8+ T lymphocytes when anti-CD19 CAR+ T lymphocytes are incubated with K562-CD19 effector cells in the presence of lenalidomide or an alternative kinase-targeting compound.

[0157] Figure 15B shows levels of CD25+ expression in T lymphocytes, both CD4+ and CD8+, when anti-CD19 CAR+ T lymphocytes are incubated with PD-1 effector cells in the presence of lenalidomide or an alternative kinase-targeting compound.

[0158] Figure 16 shows the amount of IL-10 in culture supernatants after incubation of anti-CD19 CAR+ T lymphocytes with K562-CD19 effector cells at an effector to target cell ratio (E:T) of 3:1 or 9:1, in the presence of or in the absence of various concentrations of lenalidomide.

[0159] Figure 17A shows the fold increase in cell numbers after stimulation of CAR+ T lymphocytes with anti-CD19 from two donors (pt 1 and pt 2) with K562-CD19 effector cells in the presence or absence of 1 μM lenalidomide or 50 nM or 500 nM alternative kinase-targeting compound. Figv shows the number of doublings of cells compared with the initial number after the 2nd and 4th stimulation.

[0160] Figure 18A shows the cytolytic activity of anti-CD19 CAR+ T-lymphocytes from two donor cells (pt 1 and pt 2) restimulated with K562-CD19 cells (labeled with NucLight Red (NLR)) and in the presence of 1 μM lenalidomide or 50 nM or 500 nM alternative compound targeting the kinase.

[0161] FIG. 18B shows the percent killing of target cells by anti-CD19 CAR+ T-lymphocytes from two donor cells (1 or 2) restimulated with K562-CD19 cells compared to a vehicle-only control (set at 100% ).

[0162] FIG. 19A shows a histogram of CTV cell staining, in general, in a CAR+ anti-BCMA T-lymphocyte composition after incubation with beads (200 μg/ml BCMA-conjugated beads composition) at a ratio of T-lymphocytes to beads of 1:1 and in the presence or absence of 5 μM lenalidomide.

[0163] FIGS. 19B and 19C show flow cytometry histograms for CD25 in CD4+ T lymphocytes (left panel) or CD8+ T lymphocytes (right panel) present in the CAR+ anti-BCMA? T lymphocyte composition. after incubation with beads (200 μg/ml composition BCMA-conjugated beads) at a ratio of T-lymphocytes to beads or immobilized anti-CD3 1:1, respectively, in the presence or absence of lenalidomide.

[0164] Figures 20A-20I show graphs depicting the levels of transcription factors and activation markers in CD4+ T lymphocytes (left panels) or CD8+ T lymphocytes (right panels)? present in or on anti-BCMA CAR+ T cell composition, after incubation without stimulation or with varying amounts of BCMA-conjugated beads or anti-CD3 and anti-CD28 conjugated beads and in the presence of 0 μM, 0.5 μM, or 50 μM lenalidomide . The levels of Blimp1 (Fig. 20A), CD25 (Fig. 20B), CD31 (Fig. 20C), PD-1 (Fig. 20D), Tbet (Fig. 20E), EOMES (Fig. 20F), GATA3 (Fig. 20G), Helios (Fig. 20H) and Ikaros (Fig. 20I). 200 BCMA, 50 BCMA, and 5 BCMA show BCMA-conjugated beads generated by incubating BCMA with beads of 200, 50, and 5 μg BCMA for approximately 4×10 ⁸ beads, respectively.

[0165] Figures 21A-C show graphs depicting the levels of extracellular IFN-gamma (Figure 21A), IL-2 (Figure 21B), and TNF alpha (Figure 21C) from cultures after incubation of a CAR+ anti- BCMA with two different numbers of BCMA-conjugated beads? in the presence or absence of 5 μM lenalidomide. 50 µg BCMA and 5 µg BCMA show BCMA-conjugated beads generated by incubating BCMA with 50 and 5 µg BCMA beads for approximately 4×10 ⁸ beads, respectively.

[0166] Figure 21D shows a graph depicting extracellular IL-2 levels from cultures after incubation of an anti-BCMA CAR+ T-lymphocyte composition from two different donors with different amounts of BCMA-conjugated beads, in the presence of 0 μM, 1 μM, or 5 μM lenalidomide. 200 BCMA and 5 BCMA show BCMA-conjugated beads generated by incubating BCMA with beads at 200 μg and 5 μg BCMA for approximately 4×10 ⁸ beads, respectively.

[0167] Fig. 21E and Fig. 21F show total cell counts after culturing a CAR+ anti-BCMA T cell composition after incubation for 4 days (FIG. 21E) or 7 days (FIG. 21F) with varying amounts of BCMA-conjugated beads in the presence of 5 μM lenalidomide. 50 BCMA and 5 BCMA show BCMA-conjugated beads generated by incubating the BCMA antigen with 50 µg and 5 µg BCMA beads for approximately 4 x 10^eight balls, respectively.

[0168] FIG. 21G shows histograms of CTV staining of CD4+ T lymphocytes or CD8+ T lymphocytes in anti-BCMA CAR+ T lymphocyte composition after incubation for 4 or 7 days with BCMA conjugated beads in the presence of 5 μM lenalidomide or in the absence of lenalidomide (carrier medium).

[0169] FIGS. 21H and 21I show graphs depicting the percentage of cells positive for the EGFRt surrogate marker, as determined by an anti-EGFR antibody, after incubating a CAR+ anti-BCMA T cell composition for 4 days (FIG. 21H) or 7 days (FIG. 21I) with varying amounts of BCMA-conjugated beads in the presence of 5 μM lenalidomide or in the absence of lenalidomide (carrier medium). “50” and “5” show beads generated by incubating BCMA with beads, at 50 μg and 5 μg BCMA for approximately 4×10 ⁸ beads, respectively.

[0170] FIG. 21J shows percent cell killing for target RPMI-8226 cells by CAR+ anti-BCMA effector T lymphocytes that are incubated with various amounts of BCMA-conjugated beads, in the presence of 5 μM lenalidomide or in the absence of lenalidomide (vehicle medium) . The cytolytic activity of compositions containing a ratio of effector cells to target cells of 3:1 or 1:1 and in the presence or absence, in the future, of lenalidomide is shown. “50” and “5” show BCMA-conjugated beads generated by incubating BCMA with beads of 50 and 5 μg BCMA for approximately 4×10 ⁸ beads, respectively.

[0171] Figure 22A shows a flow cytometric analysis of phosphorylated STAT5 after 2 hours of CAR stimulation (stim) with 50 μg BCMA beads. The absence of control stimulation is indicated by the dotted line. 22B shows flow cytometric analysis of intracellular cytokine levels on a representative normal T CAR donor after 24 hours of BCMA bead stimulation (tuned to transduced live CD3+).

[0172] FIGS. 23A-23B show the results of a sequential restimulation assay of anti-BCMA CAR T-lymphocyte compositions that are incubated for seven days with BCMA-conjugated beads (50 μg/ml). The results of compositions from three different donors are shown. Fig.23A and Fig.23B show the cytolytic activity of T-lymphocytes CAR+ anti-BCMA at each time point for two different donors.

[0173] Figure 24A shows results for antigen-specific CAR cytolytic activity and Figure 24B shows results for cytokine production for anti-BCMA CAR T lymphocytes that are pre-stimulated with BCMA beads (versus freshly thawed (not pre-stimulated) T -lymphocytes CAR anti-BCMA) in cocultures, comparing cells cultured in the presence and absence of lenalidomide. 24C shows overall viability and cell counts assessed for three anti-BCMA T CAR donors. 24D shows the results of flow cytometric analysis of CD25 and PD-1 surface expression (mean fluorescence intensity (MFI), for CD4+ or CD8+ CAR anti-BCMA T-lymphocytes after stimulation (pre-treatment) with BCMA beads for 7 days, in the presence of or in the absence of 1 μM lenalidomide Figure 24E shows flow cytometric analysis among CAR T donors for median fluorescence intensity (MFI; CD25 and Tim3) or the percentage of positive PD-1 and Lag3 markers on the surface of T lymphocytes in the CD4+ CAR+ and CD8+ CAR+ subsets (set on live CD3+ cells) Values shown are MFI baseline percent (Veh), viability or counts.

[0174] Figure 25A shows an analysis of effector cytokine production following CAR-specific stimulation with 50 μg BCMA beads for 24 hours in the presence of 1 μM lenalidomide compared to baseline (vehicle) response for each of the three donors.

[0175] Figure 25B shows the effects of anti-BCMA CAR T lymphocytes activated at various concentrations of BCMA beads (i.e., 5 μg, 50 μg, and 200 μg), in the presence or absence of lenalidomide (0.1 μM or 1 μM ) on the production of CAR T effector cytokines.

[0176] Figure 25C shows cytokine production by anti-BCMA CAR T lymphocytes derived from representative healthy donors and multiple myeloma patients stimulated with BCMA beads with or without the addition of PD-L1 to the beads, in the presence of 1 μM lenalidomide) or in the absence of lenalidomide.

[0177] Figures 26A and 26B show principal component analysis (PCA) results for gene expression (based on RNA-seq results; Figure 26A) and chromatin availability (based on ATAC-seq results; Figure 26B), in CAR-expressing Anti-BCMA T-lymphocytes generated by 4 different donors (Donors 1-4), stimulated with BCMA-conjugated beads for 24 hours (24 hours + stim) or 7 days (d7 + stim), or cultured without stimulation for 24 hours (24 hours), in the presence or absence of lenalidomide.

[0178] Figures 27A and 27B show volcano plots depicting the statistical significance of expression (log ₁₀ p-value) with log ₂ -fold change in gene expression, including genes or peaks that show increased (right side) or decreased (left side) expression in CAR+ T-lymphocytes stimulated with BCMA-conjugated beads for 24 hours (24 hours+stim, Fig. 27A) or 7 days (d7+stim, Fig. 27B), in the presence or absence of lenalidomide. The tables show the number of genes or peaks that show a statistically significant increase (up) or decrease (down) in expression.

[0179] Figures 27C and 27D show volcano plots depicting the statistical significance of an expression (established log ₁₀ p-value) with a log ₂ fold change in G chromatin availability, including genes or peaks that show increased (right side) or decreased (left side ) availability, in CAR+ T-lymphocytes stimulated with BCMA-conjugated beads, within 24 hours (24 hours+stim, Fig. 27C) or 7 days (d7+stim, Fig. 27D). The tables show the number of genes or peaks that show a statistically significant increase (up) or decrease (down) in availability.

[0180] Figures 28A and 28B show the direction and significance of expression for genes in biological signaling pathways that are enriched in sets of genes for which expression is statistically significantly up or down in CAR+ T lymphocytes stimulated with BCMA-conjugated beads over 24 hours (24 hours+stim, Fig. 28A) or 7 days (d7+stim, Fig. 28B).

[0181] Figure 29 shows a graph comparing individual chromatin availability peaks (diamonds) and changes in mean chromatin availability for each gene (circles), as gene expression changes, selected genes are involved in T-lymphocyte activation and signaling.

[0182] Figure 30 shows analysis of motif enrichment, enrichment log p-values, abundance, and transcription factors predicted to associate motifs for increased availability peaks in the presence of lenalidomide in day 7 cultures.

[0183] Figure 31 shows a flow cytometric analysis of intracellular expression of Ikaros in both CAR-expressing CD4+ anti-CD19 T lymphocytes and CAR-expressing CD8+ anti-CD19 T lymphocytes. CAR-expressing T lymphocytes are stimulated with CAR-T anti-idiotypic antibody (5 μg/mL) treated over a range of concentrations of lenalidomide or Compound 1. Median fluorescence intensity (MFI) values for Ikaros are normalized and calculated as a percentage of the control carrier. environment.

[0184] Figures 32A and 32B show an analysis of cytokine production by CAR-expressing anti-CD19 T lymphocytes in the presence of Compound 1 (FIG. 32A) or lenalidomide (FIG. 32B) after incubation with target cells. Multiplex cytokine analysis of supernatants performed 24 hours later from triple wells of CAR-expressing anti-CD19 T-lymphocytes co-cultured with target K562.CD19 cells in the presence of several concentrations of Compound 1 or lenalidomide. Determine the concentration of IFN-γ, IL-2 and TNF-α for CAR-expressing T-lymphocytes from three different donors for two E:T ratios. Data are mean +/- S.D. (standard deviation) for 3 experiments.

[0185] Fig.33 shows the analysis of the cytolytic function of CAR-expressing anti-CD19 T-lymphocytes in the presence of Compound 1 or lenalidomide after incubation with target cells. CAR-expressing anti-CD19 T lymphocytes from three different donors were co-cultured with target K562.CD19 cells in triplicate for two E:T ratios in the presence of Compound 1 or lenalidomide for 5 days. The results are calculated as a normalized death rate. Data are mean +/- S.D. for 3 experiments.

[0186] Figures 34A and 34B show an analysis of cytokine production by CAR-expressing anti-CD19 T lymphocytes in the presence of Compound 1 (FIG. 34A) or lenalidomide (FIG. 34B) stimulated with anti-idiotypic antibodies. Multiplex cytokine analysis of supernatants was performed 24 hours later from triple wells of anti-CD19 CAR-expressing T-lymphocytes co-cultured with an anti-idiotypic antibody agonist in the presence of 100 or 1000 nM Compound 1 (FIG. 34A), or 500 or 5000 nM lenalidomide (FIG. .34B). Determine the concentration of IFN-γ, IL-2 and TNF-α for CAR-expressing T-lymphocytes from three different donors. Data are mean +/- S.D. for 3 experiments.

[0187] Figures 35A and 35B show an analysis of surface marker expression on CAR-expressing CD4+ anti-CD19 T lymphocytes (FIG. 35A) and CAR-expressing CD8+ anti-CD19 T lymphocytes (FIG. 35B) in the presence of Stimulating Compound 1 anti-idiotypic antibodies. CAR-expressing anti-CD19 T lymphocytes from three different donors are stimulated with anti-idiotypic antibody at 0, 0.3, 3, or 30 μg/ml in the presence of 100 or 1000 nM Compound 1. Cells are analyzed by flow cytometry on day 4 Calculate the absolute change in median fluorescence intensity with respect to vehicle control for each concentration of anti-idiotypic antibody. Data are presented for 3 experiments.

[0188] Figures 36A and 36B show an analysis of surface marker expression on CD4+ anti-CD19 CAR-expressing T lymphocytes (FIG. 36A) and CD8+ anti-CD19 CAR-expressing T lymphocytes (FIG. 36B) in the presence of lenalidomide after anti-CD19 stimulation. - idiotypic antibodies. CAR-expressing anti-CD19 T lymphocytes from three different donors are stimulated with anti-idiotypic antibody at 0, 0.3, 3 or 30 μg/ml in the presence of 500 or 5000 nM lenalidomide. Cells are analyzed by flow cytometry on day 4. The absolute change in median fluorescence intensity relative to vehicle control for each concentration of anti-idiotypic antibody is calculated. Data are from 3 experiments.

[0189] Figures 37A and 37B show an analysis of CD28 surface expression on CAR-expressing CD4+ and CD8+ anti-CD19 T lymphocytes in the presence of Compound 1 (FIG. 37A) or lenalidomide (FIG. 37B) after sequential stimulation. CAR-expressing anti-CD19 T lymphocytes from three different donors stimulate K562.CD19 at an E:T ratio of 2.5:1 every 3-4 days in the presence of Compound 1 (FIG. 37A) or lenalidomide (FIG. 37B). The percentage of cells positive for CD28 is measured by flow cytometry on day 28.

[0190] Fig.38 shows the analysis of the cytolytic function of CAR-expressing anti-CD19 T-lymphocytes in the presence of Compound 1 or lenalidomide after sequential stimulation. CAR-expressing anti-CD19 T lymphocytes from three different donors, after 24 consecutive days of stimulation, are co-cultured with irradiated target K562.CD19 cells in triplicate at two E:T ratios in the presence of Compound 1 or lenalidomide. The results are calculated as a normalized death rate.

[0191] Figures 39A and 39B show an analysis of population doublings of anti-CD19 CAR-expressing T lymphocytes during a 28-day sequential stimulation period in the presence or absence of Compound 1. Anti-CD19 CAR-expressing T lymphocytes from three different donors are stimulated target K562.CD19 cells at an E:T ratio of 2.5:1 or 10:1 every 3-4 days in the presence of 500 nM Compound 1 for 28 days (represented by the x-axis). Cells are counted after each stimulation and cell doubling is calculated. (FIG. 39A) The percentage change in cell doublings on day 24 of sequential stimulation in the presence of 10 nM, 100 nM, or 500 nM Compound 1 is shown in FIG. 39B. Data represent the mean +/-S.E.M (standard error of the mean) of triplicate treatments from 3 donors. Each arrow represents a time point in restimulation.

[0192] Figures 40A and 40B show an analysis of population doublings of anti-CD19 CAR-expressing T-lymphocytes during a 28-day sequential stimulation period in the presence or absence of lenalidomide. CAR-expressing anti-CD19 T lymphocytes from three different donors are stimulated with target K562.CD19 cells at an E:T ratio of 2.5:1 or 10:1 every 3-4 days in the presence of 1000 nM lenalidomide for 28 days (shown in Figure 1). x axis). Cells are counted after each stimulation and cell doubling is calculated. (FIG. 40A) The percentage change in cell doublings on day 24 of sequential stimulation in the presence of 100 nM or 1000 nM lenalidomide is shown in FIG. 40B. Data represent the mean +/-S.E.M of triplicate treatments from three donors. Each arrow represents a time point in restimulation.

Detailed description

[0193] Provided herein is a combination therapy comprising the administration of an immunotherapy involving T-lymphocyte function or activity, such as T-cell therapy, and an immunomodulatory compound, such as a structural or functional analog or derivative of thalidomide, and/or an inhibitor of E3 ubiquitin ligase . In some aspects, the present methods enhance or modulate the proliferation and/or activity of T lymphocytes associated with the administration of immunotherapy or an immunotherapeutic agent, such as a composition containing cells for adoptive cell therapy, such as, for example, T cell therapy (e.g., CAR -expressing T-lymphocytes). In some such embodiments, the combination therapy comprises administering an immunomodulatory compound, such as a structural or functional analog of thalidomide and/or an E3 ubiquitin ligase inhibitor, and administering a T cell therapy, such as a composition containing cells for adoptive cell therapy, such as, for example, T-cell therapy (for example, CAR-expressing T-lymphocytes).

[0194] T-lymphocyte-based therapy, such as adoptive T-cell therapy (including therapy involving the introduction of cells expressing chimeric receptors specific for the disease or disorder of interest, such as chimeric antigen receptors (CARs) and/or other recombinant antigen receptors, as well as other types of adoptive immune cell and adoptive T-cell therapies) may be effective in the treatment of cancer and other diseases and disorders. The engineered expression of recombinant receptors, such as chimeric antigen receptors (CARs), on the surface of T lymphocytes makes it possible to redirect the specificity of T lymphocytes. In clinical studies, CAR T lymphocytes, such as anti-CD19 CAR T lymphocytes, produce persistent complete responses in patients with both leukemia and lymphoma (Porter et al. (2015) Sci Transl Med., 7:303ra139 ; Kochenderfer (2015) J. Clin. Oncol., 33: 540-9; Lee et al. (2015) Lancet, 385:517-28; Maude et al. (2014) N Engl J Med, 371:1507-17 ).

[0195] In a certain context, available approaches to adoptive cell therapy may not always be completely satisfactory. In some context, optimal performance may depend on the ability of the injected cells to recognize and bind the target, eg, the target antigen, to locate and successfully enter the appropriate centers in the subject, tumors and their environment. In some context, optimal efficacy may depend on the ability of the injected cells to activate, proliferate, perform various effector functions, including cytotoxic killing and secretion of various factors such as cytokines, persistence, including long-term, differentiation, transition or entry when reprogrammed into certain phenotypic conditions (such as long-lived memory, less differentiated, and effector conditions) to eliminate or reduce immunosuppressive conditions in the local disease microenvironment, to ensure efficient and robust re-responses after interruption and re-exposure for the target ligand or antigen, and to prevent or reduce exhaustion, weakness, peripheral tolerance, eventual differentiation and/or differentiation into a suppressive state.

[0196] In some embodiments, the exposure and persistence of the genetically engineered cells decrease or fall after administration to a subject. Furthermore, the observation indicates that, in some cases, increased exposure of the subject to administered cells expressing recombinant receptors (eg, increased cell number or time duration) may improve the efficacy and therapeutic outcomes of adoptive cell therapy. Preliminary analysis following administration of various CD19-targeted CAR-expressing T-lymphocytes to subjects with various CD19-expressing cancers in a variety of clinical studies proves a correlation between greater and/or longer exposure for CAR-expressing cells and treatment outcomes. Such results include patient survival and remission, even in individuals with an acute stage or significant tumor mass.

[0197] In some aspects, the proposed methods and applications provide or achieve improved or more robust responses or efficacy compared to certain alternative methods, for example, in specific groups of treated subjects. In some embodiments, the methods are advantageous by administering a T cell therapy, such as a composition containing cells for adoptive cell therapy, such as, for example, T cell therapy (e.g., CAR expressing T lymphocytes), and an immunomodulatory compound , such as a structural or functional analogue or derivative of thalidomide and/or an inhibitor of E3 ubiquitin ligase, for example, lenalidomide.

et al. (2016) Oncoimmunology, April; 5(4): e1115940).[0198] The proposed methods are based on the observation that an immunomodulatory compound, such as a structural or functional analogue or derivative of thalidomide and/or an inhibitor of E3 ubiquitin ligase, for example, lenalidomide, improves the functioning of T-lymphocytes, including functions related to reproduction, proliferation and persistence T-lymphocytes. Lenalidomide is an immunomodulatory drug currently approved for the treatment of multiple myeloma (MM) and mantle cell lymphoma (MCL) and has been clinically studied in the treatment of diffuse large B-cell lymphoma, an immunophenotype with activated B-lymphocytes. In some cases, lenalidomide enhances antitumor immune responses, at least in part, by modulating the activity of the E3 ubiquitin ligase-cereblon complex (CRBN), resulting in increased ubiquitinylation of the transcription factors Ikaros and Aiolos, which in turn results in altered expression of various receptors. on the surface of tumor cells (see, for example,

et al. (2016) Oncoimmunology, April; 5(4): e1115940).

[0199] The present data show that combination therapy with an immunomodulatory compound, such as a structural or functional analogue or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor, e.g. lenalidomide, in methods involving T lymphocytes, e.g. therapy, achieves an improvement in the function of T-cell therapy. In some embodiments, the combination of cell therapy (e.g., administration of engineered T lymphocytes) with an immunomodulatory compound, e.g., lenalidomide, improves or enhances one or more of the functions and/or effects of T cell therapy, such as persistence, proliferation, cytotoxicity. , and/or therapeutic results, for example, the ability to destroy or reduce the mass of a tumor or other diseased or target cell.

[0200] In specific aspects, herein, an immunomodulatory compound, such as a structural or functional analog or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor, such as lenalidomide, is found to promote continued function and/or survival of cells from T cell therapy (for example, CAR T-lymphocytes) after activation, including after encountering an antigen. In some aspects, lenalidomide enhances the persistence or long-term capacity of such T lymphocytes, for example by preventing cell depletion or death. In some embodiments, such improvements may result in a combination therapy demonstrating improved overall responses, e.g., reduction in tumor mass, and/or improved survival compared to subjects treated with a monotherapy comprising administration of T cell therapy alone (e.g., , CAR T-lymphocytes) or an immunomodulatory compound (eg, lenalidomide). In some aspects, the proposed methods increase overall response and/or survival greater than 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more , compared with alternative treatment, for example, compared with monotherapy, including the introduction of one T-cell therapy (eg, CAR T-lymphocytes) or an immunomodulatory compound (eg, lenalidomide).

[0201] In some embodiments, the combination with an immunomodulatory compound although improves one or more outcomes or functional attributes, does not affect one or more side effects or undesirable changes in T-lymphocytes, for example, does not reduce the ability of cells to activate, secrete one or more desired cytokines, proliferate and/or persist, for example, as measured by an in vitro assay compared to such cells cultured under otherwise the same conditions but in the absence of the immunomodulatory compound. Thus, in some embodiments, methods and combinations are provided that result in an improvement in T-lymphocyte function or phenotype, e.g., intrinsic T-lymphocyte functionality and/or intrinsic T-lymphocyte phenotype, generally without impairing one or more other desired properties or functionality, for example, the functionality of CAR T-lymphocytes.

[0202] In some embodiments, the proposed methods can enhance T-cell therapy, for example, therapy with CAR T-lymphocytes, which, in some aspects, can improve treatment outcomes. In some embodiments, these methods are particularly advantageous for subjects in which cells from T cell therapy show poor proliferating, deplete, show decreased or decreased persistence in a subject, and/or subjects who have a cancer that is persistent or refractory. treatment with other therapies is an aggressive or high-risk cancer and/or that shows or is likely to show a relatively lower proportion of response to CAR T-lymphocyte therapy administered without an immunomodulatory compound compared to the other type cancer or compared with the introduction of another therapy with CAR T-lymphocytes.

[0203] In some aspects, the proposed methods can improve, increase or enhance T-cell therapy so as to overcome the lack of persistence and / or depletion of T-lymphocytes, for example, in subjects that correspond or approximately correspond to days 12-15 after the start administration of T-cell therapy, less than 10 μl, for example, less than 5 μl or less than 1 μl of such cells, or their CD8+ or CD3+ subset, are detected in the blood. In some embodiments, a subject receiving an administration of a T cell therapy, e.g., CAR T cells, is monitored for the presence, absence, or level of T cells from the therapy in the subject, e.g., in a biological sample from the subject, e.g., in the subject's blood. In some embodiments, an immunomodulatory compound, such as a structural or functional analogue or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor, e.g., lenalidomide, is administered to a subject receiving T cell therapy (e.g., CAR T lymphocytes), but has proliferate weakly and/or are at or below a threshold level in a sample from a subject, e.g., in a blood sample, while strong or sustained proliferation of CAR T lymphocytes in a subject is typically seen in multiple subjects receiving T-cell therapy (eg, CAR-T), in some cases, the same T-cell therapy (eg, the same CAR T-lymphocytes). In some aspects, an immunomodulatory compound, such as a structural or functional analog or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor, such as lenalidomide, is administered if, on or about day 12-15 after initiation of T cell therapy, blood is detected less than 10 μl, for example, less than 5 μl or less than 1 μl of such cells, or a subset of CD8+ or CD3+.

[0204] In certain aspects, the proposed methods can improve, increase or enhance T-cell therapy in subjects who have a peak response to T-cell therapy, but who have a response, for example, the presence of T-lymphocytes and / or weight loss tumor is reduced or no longer detected. In some aspects, an immunomodulatory compound, such as a structural or functional analog or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor, such as lenalidomide, is administered to a subject within one week, such as within, 1, 2, or 3 days after: ( i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0205] In some embodiments, the methods can be used to treat a disease or condition, e.g., B-lymphocyte cancer or hematological malignancy, and in particular, diseases, conditions, or malignancies in which responses, e.g., complete response, to treatment with T-cell therapy alone, such as a composition containing cells for adoptive cell therapy, such as T-cell therapy (eg, CAR-expressing T-lymphocytes), is relatively low compared to treatment with other types of T cell therapy or treatment of other diseases or malignancies (e.g., CR in less than or less than about 60%, less than about 50%, or less than about 45% of subjects so treated) and/or in which the subject does not respond to treatment with an immunomodulatory compound such as a structural or functional analog or derivative of thalidomide and/or an E inhibitor 3 ubiquitin ligases, such as lenalidomide, by itself.

[0206] In some embodiments, the combination therapy provided herein is for use in a subject having a cancer who, upon initiation of T cell therapy, such as a composition containing cells for adoptive cell therapy, e.g., CAR expressing T-lymphocytes, the subject relapses into remission following treatment with T-cell therapy. In some embodiments, subjects who relapse after such remission are administered an immunomodulatory compound, such as a structural or functional analog or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor, such as lenalidomide. In some embodiments, the combination therapy provided herein is for use in a subject having a disease or condition, such as cancer, in which the amount of immunomodulatory compound administered is deficient, as a single agent and/or without administration of a T cell therapy, to alleviate, reduce or prevent the disease or condition or symptom or result, for example, is insufficient to alleviate, reduce or prevent the disease or condition in the subject or symptom or result. In some embodiments, the method thereby reduces or alleviates the symptom or outcome or severity of the disease or condition to an extent that is greater than a combination of (i) the amount of reduction or alleviation achieved by administering the immunomodulatory agent alone, optionally the average of the subject population having the disease or condition, and (ii) the degree of reduction or amelioration by the administration of T cell therapy alone, optionally, the average for the population of subjects having the disease or condition. In some embodiments, the method reduces or alleviates such symptoms, treatment outcomes, or disease severity, e.g., compared to the mean for a population of subjects having the disease or condition greater than or greater than about 1.5-fold, 2, 0x, 3.0x, 4.0x, 5.0x, 6.0x, 7.0x, 8.0x, 9.0x, 10.0x times, 20.0 times, 30.0 times, 40.0 times, 50.0 times or more.

[0207] In some embodiments, the proposed combination therapy is used in combination with the treatment of certain diseases or conditions, for example, cancer, in which optimal stimulation of the recombinant antigen receptor, for example, CAR T-lymphocytes, is difficult to achieve and / or is not observed constantly. In some embodiments, less than optimal stimulation may result from low or unavailable levels of the disease antigen in vivo, eg, in or on a tumor. In some embodiments, certain cancers, such as NHL, e.g., high-risk NHL or aggressive NHL, such as DLBCL and/or chronic lymphocytic leukemia (CLL), may be associated with defects or a decrease in intrinsic T-lymphocyte functionality, by which, in some cases, is affected by the disease itself. For example, the pathogenesis of many cancers, such as CLL and NHL, e.g., DLBCL, may be associated with immunodeficiency resulting in facilitation of tumor growth and immune evasion, e.g. tumors. In some cases, alleviating intrinsic defects of T lymphocytes derived from such patients' cancers for use in combination with adoptive cell therapy could provide stronger responses to adoptive T cell therapy, such as CAR T cell therapy. In some cases, less than optimal stimulation may be caused by differences in the level of CAR expression on the GE T lymphocytes administered to the subject. In any such embodiment, administration of an immunomodulatory compound, such as a structural or functional analog or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor, such as lenalidomide, may improve the stimulation or activity of such T lymphocytes in vivo in a subject.

[0208] In some embodiments of the proposed methods, one or more properties of the injected genetically engineered cells can be improved or increased or made more compared to the input cells of the reference composition, for example, to increase or prolong the multiplication and / or persistence of such input cells in a subject or increase or increase the evoked response when re-stimulated with the antigen. In some embodiments, the increase may be at least 1.2-fold, at least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, or at least A 10-fold increase in such a property or feature compared to the same property or feature when a reference cell composition is administered. In some embodiments, an increase in one or more of these properties or features can be observed or is present within 7 days, 14 days, 21 days, within one month, two months, three months, four months, five months, six months, or 12 months after administration of the engineered cells and initiation of administration of an immunomodulatory compound, such as a structural or functional analogue or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor, for example, lenalidomide.

[0209] In some embodiments, the reference cell composition may be a composition of T lymphocytes from the blood of a subject who does not have or is expected to have no cancer, or is a population of T lymphocytes obtained, isolated, generated, produced, incubated and/or administered under the same or substantially the same conditions, except that they are not incubated or administered in the presence of an immunomodulatory compound. In some embodiments, the reference cell composition contains engineered cells that are essentially the same, including expressing the same recombinant receptor, such as CAR. In some aspects, such T-lymphocytes are processed identically or substantially identically, for example, they are produced in a similar way, prepared in a similar way, administered at the same or about the same dose level, and other similar factors.

[0210] In some embodiments, the proposed methods result in engineered cells with improved persistence and/or greater potency in the subject to which they are administered. In some embodiments, the persistence of engineered cells, such as CAR-expressing T-lymphocytes, in a subject is greater than what could be achieved with alternative methods, such as methods comprising administering a reference cell composition, for example, the introduction of T-cell therapy, but without the introduction of an immunomodulatory compound. In some embodiments, persistence is increased at least, or about, at least 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8 times, 9 times, 10 times, 20 times, 30 times, 50 times, 60 times, 70 times, 80 times, 90 times, 100 times or more.

[0211] In some embodiments, the degree or level of persistence of administered cells can be detected or quantified after administration to a subject. For example, in some aspects, quantitative PCR (qPCR) is used to evaluate the number of cells expressing the recombinant receptor (eg, CAR expressing cells) in the blood or serum or organ or tissue (eg, disease center) of a subject. In some aspects, quantification persistence is defined as the number of copies of a DNA or plasmid encoding a receptor, e.g., CAR, per microgram of DNA, or as the number of receptor-expressing, e.g., CAR-expressing cells per microliter of a sample, e.g., blood or serum, or relative to total peripheral blood mononuclear cells (PBMC) or leukocytes or T-lymphocytes per microliter of sample. In some embodiments, flow cytometric analysis can also be performed to detect cells expressing the receptor, typically using antibodies specific for those receptors. Cell-based assays can also be used to detect the number or percentage of functional cells, such as cells capable of binding and/or neutralizing and/or inducing reactions, e.g., cytotoxic reactions, against cells of a disease or condition or expressing an antigen recognized by a receptor. In any such embodiment, the extent or level of expression of another marker associated with the recombinant receptor (eg, CAR-expressing cells) can be used to distinguish the introduced cells from endogenous cells in the subject.

[0212] Also provided are methods of genetic engineering, preparation and production of cells, compositions containing cells and/or immunomodulatory compound and kits, and devices containing cells and/or immunomodulatory compounds and intended for use, production and administration of cells and/or immunomodulatory compound , for example, together with the proposed methods of combination therapy.

[0213] All publications, including patent documents, scientific articles and databases referred to in this application, are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication were separately incorporated by reference. If the definition given herein conflicts with or otherwise does not match the definition given in patents, applications, published applications and other publications, which are incorporated herein by reference, the definition given in this document shall take precedence over the definition that is incorporated herein by reference.

[0214] The section headings used herein are for organizational purposes only and are not intended to limit the subject matter described.

I. Combined therapy

[0215] Provided herein are methods of combination therapy for treating a disease or disorder, such as a cancerous or proliferative disease, which comprises administering to a subject a combination therapy of 1) an immunomodulatory compound such as a structural or functional analog or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor , for example, lenalidomide, and 2) T-cell therapy, for example, CAR-expressing cells, for example, T-lymphocytes. In some embodiments, the T cell therapy is an adoptive immunocellular therapy comprising T lymphocytes that specifically recognize and/or target an antigen associated with a disease or disorder, such as a cancerous or proliferative disease. Also provided are combinations and articles of manufacture, such as kits, that contain a composition containing a T cell therapy and/or a composition containing an immunomodulatory compound, and uses of such compositions and combinations for the treatment or prevention of diseases, conditions, and disorders, including cancers.

[0216] In some embodiments, such methods may include administering an immunomodulatory compound, such as a structural or functional analog or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor, e.g., lenalidomide, before, simultaneously, during, during a course (including once and / or periodically during the course), and / or after the introduction (for example, the beginning of the introduction) of T-cell therapy (for example, CAR-expressing T-lymphocytes). In some embodiments, administration may include sequential or alternating administration of an immunomodulatory compound and T cell therapy.

[0217] In some embodiments, the cell therapy is an adoptive cell therapy. In some embodiments, the cell therapy is or includes tumor infiltrating lymphocyte (TIL) therapy, transgenic TCR therapy, or therapy with cells expressing a recombinant receptor (optionally, T-cell therapy), which is optionally a therapy with cells, expressing a chimeric antigen receptor (CAR). In some embodiments, the therapy is a B-lymphocyte-targeting therapy. In some embodiments, the therapy targets B-lymphocyte maturation antigen (BCMA). In some embodiments, the therapy targets CD19. In some embodiments, the cells and dosing regimens for administering the cells may include any of the cells and dosing regimens as described in the following subsection A, titled "Administration of T Cell Therapy".

[0218] In some embodiments, the immunomodulatory compound enhances the functionality of T-lymphocytes. In some embodiments, the immunomodulatory compound elicits anti-myeloma activity. In some embodiments, the immunomodulatory compound alters the suppressive microenvironment. In some embodiments, the immunomodulatory compound is a structural or functional analog or derivative of thalidomide. In some embodiments, the immunomodulatory compound is an E3 ubiquitin ligase inhibitor. In some embodiments, the immunomodulatory compound is lenalidomide or a compound with the same or similar properties as lenalidomide, including analogues or derivatives, a stereoisomer of lenalidomide, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, clathrate, or polymorph thereof. In some embodiments, dosing regimens for administering an immunomodulatory compound may include any of the regimens as described in the following subsection B, entitled "Administration of an immunomodulatory compound."

[0219] In some embodiments, the T cell therapy (eg, CAR expressing T lymphocytes) and the immunomodulatory compound are provided as pharmaceutical compositions for administration to a subject. In some embodiments, the pharmaceutical compositions contain therapeutically effective amounts of one or both of the combination therapy agents, eg, adoptive cell therapy T lymphocytes and an immunomodulatory compound, as described. In some embodiments, the agents are formulated for administration in separate pharmaceutical compositions. In some embodiments, any pharmaceutical compositions provided herein may be formulated in water in dosage forms appropriate for each route of administration.

[0220] In some embodiments, a combination therapy that includes administering a T cell therapy, including genetically engineered cells, such as CAR T cell therapy, and an immunomodulatory compound, is administered to a subject or patient having a disease or condition that must be treated (eg, cancer), or is at risk of developing a disease or condition (eg, cancer). In some aspects, the methods treat, for example, alleviate one or more symptoms of a disease or condition, for example, by reducing tumor mass in a cancer expressing an antigen recognized by immunotherapy or an immunotherapeutic agent, for example, recognized by genetically engineered T-lymphocytes .

[0221] In some embodiments, the disease or condition being treated can be any disease or condition in which antigen expression is associated with and/or involved in the etiology of the disease state or disorder, e.g., causes, exacerbates, or is otherwise involved to such a disease, condition or disorder. Exemplary diseases and conditions may include diseases or conditions associated with cancer or cell transformation (eg, cancer), an autoimmune or inflammatory disease, or an infectious disease, such as those caused by bacterial, viral, or other pathogens. Illustrative antigens, which include antigens associated with various diseases and conditions that can be treated, include any of the antigens described herein. In specific embodiments, a recombinant receptor expressed on genetically engineered cells from combination therapy, including chimeric antigen receptors or a transgenic TCR, specifically binds to an antigen associated with a disease or condition.

[0222] In some embodiments, the disease or condition is a tumor, such as a solid tumor, lymphoma, leukemia, a blood tumor, a metastatic tumor, or another type of cancer or tumor.

[0223] In some embodiments, the cancer or proliferative disease is a B-lymphocyte cancer or a hematologic malignancy. In some embodiments, the cancer or proliferative disease is lymphocytic leukemia (ALL), non-Hodgkin's lymphoma (NHL), or chronic lymphocytic leukemia (CLL). In some embodiments, the cancer is CLL. In some embodiments, the methods can be used to treat myeloma, lymphoma, or leukemia. In some embodiments, the methods can be used to treat non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), acute myeloid leukemia (AML), or myeloma, e.g., multiple myeloma. (MM). In some embodiments, the methods can be used to treat MM or DBCBL.

[0224] In some embodiments, the antigen associated with the disease or disorder is selected from the group consisting of ROR1, B cell maturation antigen (BCMA), tEGFR, Her2, Ll-CAM, CD19, CD20, CD22, mesothelin, CEA and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, EGP-2, EGP-4, EPHa2, ErbB2, 3, or 4, erbB dimers , EGFR vIII, FBP, FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kdr, kappa light chain, Ley antigen, L1 cell adhesion molecule, (L1-CAM), antigen, melanoma associated (MAGE)-A1, MAGE-A3, MAGE-A6, preferably expressed melanoma antigen (PRAME), survivin, EGP2, EGP40, TAG72, B7-H6, IL-13 receptor a2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE Al, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptor, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen and ant universal label-associated gene, cancer-testicular antigen, mesothelin, MUC1, MUC16, PSCA, NKG2D ligands, NY-ESO-1, MART-1, gp100, G protein-coupled 5D receptor (GPCR5D), oncofetal antigen, ROR1 , TAG72, VEGF-R2, carcinoembryonic antigen (CEA), prostate specific antigen, PSMA, Her2/neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2) , CE7, Wilms tumor 1 (WT-1), cyclin, cyclin A2, CCL-1, CD138 and pathogen-specific antigen. In some embodiments, the antigen is associated with or is a universal tag.

[0225] In some embodiments, the cancer or proliferative disease expresses BCMA. In some embodiments, the proposed methods use a T lymphocyte expressing a recombinant receptor (eg, CAR T lymphocytes) that targets BCMA.

[0226] In some embodiments, the methods can be used to treat a non-hematologic cancer, such as a solid tumor. In some embodiments, the methods can be used to treat bladder, lung, brain, melanoma (e.g., small cell lung melanoma), breast, cervical, ovarian, colorectal, pancreatic, endometrial, esophageal, kidneys, liver, prostate, skin, thyroid, or uterus. In some embodiments, the cancer or proliferative disease is pancreatic cancer, bladder cancer, colorectal cancer, breast cancer, prostate cancer, kidney cancer, hepatocellular cancer, lung cancer, ovarian cancer, cervical cancer, pancreatic cancer, rectal cancer, thyroid cancer, uterine cancer, gastrointestinal cancer, esophageal cancer, head and neck cancer, melanoma, neuroendocrine cancers, CNS cancers, brain tumors, bone cancer or soft tissue sarcoma.

[0227] In some embodiments, the disease or condition is an infectious disease or condition such as, but not limited to, viral, retroviral, bacterial, and protozoal infection, immunodeficiency, cytomegalovirus (CMV), Epstein-Barr virus (EBV) , adenovirus, BK polyomaviruses. In some embodiments, the disease or condition is an autoimmune or inflammatory disease or condition, such as arthritis, e.g., rheumatoid arthritis (RA), Type I diabetes, systemic lupus erythematosus (SLE), inflammatory bowel disease, psoriasis, scleroderma, autoimmune thyroid disease, Graves' disease, Crohn's disease, multiple sclerosis, asthma, and/or a transplant related disease or condition.

[0228] To prevent or treat a disease, the appropriate dose of an immunomodulatory compound (e.g., lenalidomide) and/or immunotherapy, e.g., T-cell therapy (e.g., CAR-expressing T-lymphocytes), may depend on the type of disease to be treated , the particular immunomodulatory compound, cells and/or recombinant receptors expressed on the cells, the severity and course of the disease, the route of administration, whether the immunomodulatory compound and/or T-cell therapy is administered for preventive or therapeutic purposes, previous therapy, frequency of administration, the subject's medical history and response to cells, and the judgment of the attending physician. Compositions and cells in some embodiments, the implementation is appropriately administered to the subject at one time or over a series of treatment sessions. Exemplary dosing regimens and time schedules of proposed combination therapy are described.

[0229] In some embodiments, the T cell therapy and the immunomodulatory compound are administered as part of an additional concomitant treatment, which may be administered simultaneously or sequentially with another therapeutic intervention, in any order. In some context, T-cell therapy, for example, engineered T-lymphocytes, such as CAR-expressing T-lymphocytes, are co-administered with another therapy close enough in time that the T-cell therapy enhances the effect of one or more additional therapeutic agents, or vice versa. In some embodiments, the cells are administered prior to one or more additional therapeutic agents. In some embodiments, T cell therapy, eg, engineered T lymphocytes, such as CAR expressing T lymphocytes, is administered after one or more additional therapeutic agents. In some embodiments, the combination therapy methods further comprise anti-lymphocyte therapy, such as administration of a chemotherapeutic agent. In some embodiments, the combination therapy further comprises administering another therapeutic agent, such as an anticancer agent, checkpoint inhibitor, or other immunomodulatory agent. Uses include the use of combination therapy in such methods and treatments and the use of such compositions in the preparation of medicaments for such combination therapy methods. In some embodiments, the methods and uses thereby cure a disease or condition or disorder, such as a cancer or proliferative disease, in a subject.

[0230] Before, during, or after the administration of immunotherapy (e.g., T-cell therapy, such as therapy with CAR T-lymphocytes) and/or immunomodulatory compound, the biological activity of the T-cell therapy is measured, for example, the biological activity of gene populations. engineered cells, in some embodiments, for example, using any number of known methods. Evaluation parameters include the ability of engineered cells to destroy target cells, persistence, and other measures of T-lymphocyte activity, for example, as measured using any usable method known in the art, such as the assays further described below in Section III. In some embodiments, the biological activity of cells, e.g., T cells, administered for T cell therapy is measured by cytotoxic cell killing assay, expression and/or secretion of one or more cytokines, proliferation, or expansion, e.g., upon restimulation antigen. In some aspects, biological activity is measured by assessing disease severity and/or clinical outcome, such as reduction in tumor mass or burden. In some embodiments, administration of one or both of the combination therapy agents and/or multiple administration of therapy may be determined based on test results before, during, during, or after administration of one or both of the combination therapy agents.

[0231] In some embodiments, the combined effect of an immunomodulatory compound in combination with a cell therapy may be synergistic compared to a treatment comprising the immunomodulatory compound alone or cell therapy alone. For example, in some embodiments, the methods provided herein result in an increase or improvement in the desired therapeutic effect, such as an increase in or improvement in the reduction or slowing of one or more symptoms associated with a cancer.

[0232] In some embodiments, the immunomodulatory compound increases the multiplication or proliferation of engineered T lymphocytes such as CAR T lymphocytes. In some embodiments, an increase in multiplication or proliferation is observed in vivo when administered to a subject. In some embodiments, the number of engineered T lymphocytes, e.g., CAR T lymphocytes, increases more or more than about 1.2-fold, 1.5-fold, 2.0-fold, 3.0-fold , 4.0 times, 5.0 times, 6.0 times, 7.0 times, 8.0 times, 9.0 times, 10.0 times or more.

A. Introduction of T cell therapy

[0233] In some embodiments of the methods, compositions, combinations, kits, and uses provided herein, combination therapy comprises administering an immune cell therapy, such as T-cell therapy (eg, CAR-expressing T-lymphocytes) to a subject. The introduction of such therapy can begin before, after or simultaneously with the introduction of one or more immunomodulatory compounds, as described.

[0234] In some embodiments, a cell-based therapy is or includes the administration of cells, such as immune cells, e.g., T-lymphocytes or NK cells, that target a molecule expressed on the surface of a lesion, such as a tumor or cancer. In some embodiments, the immune cells express the T-lymphocyte receptor (TCR) or other antigen-binding receptor. In some embodiments, the immune cells express a recombinant receptor, such as a transgenic TCR or a chimeric antigen receptor (CAR). In some embodiments, the cells are autologous to the subject. In some embodiments, the cells are allogeneic to the subject.

[0235] In some aspects, T cell therapy is or comprises tumor infiltrating lymphocytic (TIL) therapy, transgenic TCR therapy, or T cell therapy involving genetically engineered cells, such as therapy with cells expressing a recombinant receptor . In some embodiments, the recombinant receptor specifically binds to a ligand, such as a ligand associated with a disease or condition, such as associated with or expressed by tumor or cancer cells. In some embodiments, T cell therapy comprises administering engineered T lymphocytes to express a chimeric antigen receptor (CAR).

[0236] In some embodiments, the cells of the invention are expressed and/or genetically engineered to express receptors such as recombinant receptors, including receptors containing ligand-binding domains or binding fragments thereof, and T-lymphocyte receptors (TCR) and components thereof, and/or functional antigen receptors other than TCRs, such as chimeric antigen receptors (CARs). In some embodiments, the recombinant receptor contains an extracellular ligand-binding domain that specifically binds to an antigen. In some embodiments, the recombinant receptor is a CAR that contains an extracellular antigen recognition domain that specifically binds to an antigen. In some embodiments, the ligand, such as an antigen, is a protein expressed on the surface of cells. In some embodiments, the CAR is a TCR-like CAR and the antigen is a truncated peptide antigen, such as an intracellular protein peptide antigen that, like TCR, is recognized on the cell surface in the context of a major histocompatibility complex (MHC) molecule.

[0237] Genetically engineered cells, including genetically engineered cells containing recombinant receptors, are described in Section II, below. Exemplary recombinant receptors, including CAR and recombinant TCR, as well as methods of genetic engineering and introduction of receptors into cells, include those described, for example, in International Patent Applications Nos. /071154, WO2013/123061, WO2016/0046724, WO2016/014789, WO2016/090320, WO2016/094304, WO2017/025038, WO2017/173256, в публикациях заявок на патент США номера US2002131960, US2013287748, US20130149337, в патентах США №№ 6451995 , 7446190, 8252592, 8339645, 8398282, 7446179, 6410319, 7070995, 7265209, 7354762, 7446191, 8324353, 8479118 и 9765342, и в заявках на Европейский патент номер EP2537416, и/или те, которые описаны Sadelain et al., Cancer Discov ., 3(4): 388-398 (2013); Davila et al., PLoS ODN 8(4): e61338 (2013); Turtle et al., Curr. Opin. Immunol., 24(5): 633-39 (2012); Wu et al., Cancer, 18(2): 160-75 (2012). In some aspects, engineered antigen receptors include CARs as described in US Patent No: 7446190 and are described in International Patent Application Publication No: WO/2014055668 A1.

[0238] In some embodiments, the antigen is or includes an αvβ6 integrin (avb6 integrin), B-lymphocyte maturation antigen (BCMA), B7-H3, B7-H6, carbonic anhydrase 9 (CA9 also known as CAIX or G250), testicular cancer antigen, testicular cancer antigen 1B (CTAG, also known as NY-ESO-1 and LAGE-2), carcinoembryonic antigen (CEA), cyclin, cyclin A2, chemokine motif C-C ligand 1 (CCL-1), CD19, CD20 , CD22, CD23, CD24, CD30, CD33, CD38, CD44, CD44v6, CD44v7/8, CD123, CD133, CD138, CD171, chondroitin sulfate proteoglycan 4 (CSPG4), epidermal growth factor protein (EGFR), truncated epidermal growth factor protein ( tEGFR), epidermal growth factor receptor type III (EGFR vIII) mutation, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), ephrin B2, ephrin A2 receptor (EPHa2), estrogen receptor, Fc receptor like 5 (FCRL5; also known as the Fc receptor homologue 5 or FCRH5), fetal acetylcholine receptor (fetal AchR), fola t-binding protein (FBP), folate receptor alpha, ganglioside GD2, O-acetylated GD2 (OGD2), ganglioside GD3, glycoprotein 100 (gp100), glypican-3 (GPC3), G protein-coupled 5D receptor (GPCR5D), Her2/neu (erb-B2 tyrosine kinase receptor), Her3 (erb-B3), Her4 (erb-B4), erbB dimers, human melanoma-associated high molecular weight antigen (HMW-MAA), hepatitis B surface antigen, leukocyte A1 antigen human leukocyte A2 antigen (HLA-A2), IL-22 receptor alpha (IL-22Rα), IL-13 alpha 2 receptor (IL-13Rα2), kinase insert domain containing receptor (kdr), kappa light chain, L1 cell adhesion molecule (L1-CAM), L1-CAM CE7 epitope, leucine-rich insert containing family 8 element A (LRRC8A), Ley antigen, melanoma-associated antigen (MAGE)-A1, MAGE-A3 , MAGE-A6, MAGE-A10, mesothelin (MSLN), c-Met, mouse cytomegalovirus (CMV), mucin 1 (MUC1), MUC16, natural killer group 2 element D ligands (NKG2D), melan A (MART-1) , adhesion molecule neuronal cell antigen (NCAM), oncofetal antigen, preferably expressed melanoma antigen (PRAME), progesterone receptor, prostate specific antigen, prostate stem cell antigen (PSCA), prostate specific membrane antigen (PSMA), tyrosine kinase receptor orphan receptor 1 (ROR1) ), survivin, trophoblast glycoprotein (TPBG also known as 5T4), tumor-associated glycoprotein 72 (TAG72), tyrosinase-related protein 1 (TRP1, also known as TYRP1 or gp75), tyrosinase-related protein 2 (TRP2, also known as dopachromtautomerase , dopachrome delta isomerase or DCT), vascular endothelial growth factor receptor (VEGFR), vascular endothelial growth factor receptor 2 (VEGFR2), Wilms tumor 1 (WT-1), pathogen-specific or pathogen-expressed antigen, or antigen associated with universal label, and/or biotinylated molecules, and/or molecules expressed by HIV, HCV, HBV or other pathogens. The antigens targeted by the receptors in some embodiments include antigens associated with B-lymphocyte cancer, such as any of a number of known B-lymphocyte markers. In some embodiments, the antigen is or includes CD20, CD19, CD22, ROR1, CD45, CD21, CD5, CD33, kappa Ig, lambda Ig, CD79a, CD79b, or CD30.

[0239] In some embodiments, the antigen is or includes a pathogen-specific or pathogen-expressed antigen. In some embodiments, the antigen is a viral antigen (such as a viral antigen from HIV, HCV, HBV, and the like), bacterial antigens, and/or parasitic antigens.

[0240] In some embodiments, the combination therapy comprises administering to the subject cells, e.g., T-lymphocytes, expressing a recombinant receptor that specifically recognizes and/or targets an antigen associated with cancer and/or present on a universal tag. In some embodiments, the antigen recognized or targeted by T lymphocytes is ROR1, B lymphocyte maturation antigen (BCMA), carbonic anhydrase 9 (CAIX), tEGFR, Her2/neu (erbB2 tyrosine kinase receptor), L1- CAM, CD19, CD20, CD22, mesothelin, CEA and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG -40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL- 22R-alpha, IL-13R-alpha2, receptor containing kinase insertion domain (kdr), kappa light chain, Ley antigen, L1 cell adhesion molecule, (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE -A3, MAGE-A6, preferably expressed melanoma antigen (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA -AI MAGE Al , HLA-A2, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, testicular cancer antigen, mesothelin, mouse CMV , mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, G protein-coupled 5D receptor (GPCR5D), oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen ( CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms tumor 1 (WT-1), cyclin, A2 cyclin, CCL-1, CD138, optionally a human antigen to any of the above; pathogen-specific antigen.

[0241] Methods for introducing genetically engineered cells for adoptive cell therapy are known and can be used in combination with the proposed methods and compositions. For example, adoptive T cell therapy methods are described, for example, in US Patent Application Publication No. 2003/0170238, Gruenberg et al; US patent No. 4690915, Rosenberg; Rosenberg (2011) Nat Rev Clin Oncol. 8(10):577-85). See, for example, Themeli et al., (2013) Nat Biotechnol. 31(10): 928-933; Tsukahara et al., (2013) Biochem Biophys Res Commun 438(1): 84-9; Davila et al., (2013) PLoS ONE 8(4): e61338.

[0242] In some embodiments, cell therapy, such as adoptive T cell therapy, is performed using autologous transfer, in which cells are isolated and/or otherwise prepared from a subject to be treated with cell therapy, or from a sample obtained from such a subject. Thus, in some aspects, cells are obtained from a subject, such as a patient in need of treatment, and the cells, after isolation and processing, are administered to the same subject.

[0243] In some embodiments, cell therapy, for example, adoptive T cell therapy, is performed using allogeneic transfer, in which cells are isolated and / or otherwise prepared from a subject other than the subject who is to receive or who ultimately receiving cell therapy, for example, the first subject. In such embodiments, the cells are then administered to another subject, such as a second subject, of the same species. In some embodiments, the first and second subjects are genetically identical. In some embodiments, the first and second subjects are genetically similar. In some embodiments, the second subject expresses the same HLA class or supertype as the first subject.

[0244] In certain embodiments, cells, or distinct populations of cell subtypes, are administered to a subject in the range of about one million to about 100 billion cells and/or in a number of cells per kilogram of body weight, e.g., from 1 million to about 50 billion cells ( for example, about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or as many as in the range specified by any the two values discussed above), e.g., about 10 million to about 100 billion cells (e.g., about 20 million cells, about 30 million cells, about 40 million cells, about 60 million cells, about 70 million cells, about 80 million cells, about 90 million cells Approximately 10 billion cells Approximately 25 billion cells Approximately 50 billion cells, about 75 billion cells, about 90 billion cells, or as many as any two of the above values), and in some cases from about 100 million cells to about 50 billion cells (e.g., about 120 million cells, approximately 250 million cells, approximately 350 million cells, approximately 450 million cells, approximately 650 million cells, approximately 800 million cells, approximately 900 million cells, approximately 3 billion cells, approximately 30 billion cells, approximately 45 billion cells) or any value in these ranges and/or per kilogram of body weight. Doses may vary depending on the specific parameters of the disease or disorder and/or the patient and/or other treatments.

[0245] In some embodiments, for example, when the subject is a human, the dose contains less than about 1 x 10 ⁸ cells, in total, expressing the recombinant receptor (e.g., CAR), T-lymphocytes, or peripheral blood mononuclear cells (PBMC), for example, in the range from about 1 x 10 ⁶ to 1 x 10 ⁸ such cells, for example, 2 x 10 ⁶ , 5 x 10 ⁶ , 1 x 10 ⁷ , 5 x 10 ⁷ , or 1 x 10 ⁸ or such cells, in in general, or an amount in the range between any two of the above values.

[0246] Cells can be administered by any appropriate means. The cells are administered in a dosing regimen to achieve a therapeutic effect, such as reduction in tumor mass. Dosing and administration may depend in part on the timing of the administration of the immunomodulatory compound, which may be administered before, after and/or simultaneously with the start of the administration of the T cell therapy. Various dosing schedules for T cell therapy include, but are not limited to, single or multiple doses at different time points, bolus administration, and pulsed infusion.

1. Compositions and formulations

[0247] In some embodiments, a dose of cells from a T cell therapy, such a T cell therapy containing genetically engineered cells with a recombinant antigen receptor, e.g., CAR or TCR, is provided as a composition or preparation, such as a pharmaceutical composition or a drug. Such compositions can be used according to the proposed methods, for example, in the prevention or treatment of diseases, conditions and disorders.

[0248] In some embodiments, a T cell therapy, such as engineered T lymphocytes (eg, CAR T lymphocytes), is formulated with a pharmaceutically acceptable carrier. In some aspects, the choice of carrier is determined in part by the particular cell or agent and/or route of administration. Accordingly, there are many suitable preparations. For example, the pharmaceutical composition may contain preservatives. Suitable preservatives may include, for example, methyl paraben, propyl paraben, sodium benzoate and benzalkonium chloride. In some aspects, a mixture of two or more preservatives is used. The preservative or mixtures thereof are typically present in an amount of from about 0.0001% to about 2% by weight of the composition as a whole. Carriers are described, for example, in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). Pharmaceutically acceptable carriers are generally non-toxic to users at the dosages and concentrations employed and include, but are not limited to: buffers such as phosphate, citrate and other organic acid buffers; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes (eg Zn-protein complexes) and/or non-ionic surfactants such as polyethylene glycol (PEG).

[0249] Buffer agents in some aspects are included in the compositions. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. In some aspects, a mixture of two or more buffering agents is used. The buffering agent, or mixtures thereof, is typically present in an amount of from about 0.001% to about 4% by weight of the total composition. Methods for preparing pharmaceutical compositions suitable for administration are known. Illustrative methods are described in more detail, for example, in Remington: The Science and Practice of Pharmacy, Lippincott Williams &Wilkins; 21st ed. (May 1, 2005).

[0250] The formulations may contain aqueous solutions. The preparation or composition may also contain several active ingredients useful for a particular indication, disease or condition that is prevented or treated by cells or agents, where the respective activities do not adversely affect each other. Such active ingredients are conveniently present in combinations, in amounts that are effective for the intended purpose. Thus, in some embodiments, the pharmaceutical composition further comprises other pharmaceutically active agents or drugs such as chemotherapeutic agents, e.g., asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracils, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab , vinblastine, vincristine, and the like.

[0251] The pharmaceutical composition in some embodiments contains cells in an amount effective to treat or prevent a disease or condition, such as a therapeutically effective or prophylactically effective amount. Therapeutic or prophylactic efficacy is, in some embodiments, monitored by periodic evaluation of treated subjects. For repeated administration over several days or more, depending on the condition, the treatment is repeated until the desired suppression of the symptoms of the disease occurs. However, other dosing regimens may be useful and may be determined. The desired dose can be delivered by separate administration bolus of the composition, by multiple bolus administrations of the composition, or by continuous administration of the composition by infusion.

[0252] Cells can be administered using standard techniques administration, preparations and/or devices. Formulations and devices such as syringes and vials are provided for storing and administering the compositions. With respect to cells, administration may be autologous or heterologous. For example, immunoreactive cells or progenitors can be obtained from one subject and administered to the same subject or another compatible subject. Immunoreactive cells derived from peripheral blood or their progeny (eg, obtained in vivo, ex vivo or in vitro) can be administered by local injection, including catheter injection, systemic injection, local injection, intravenous injection or parenteral administration. When a therapeutic composition (eg, a pharmaceutical composition containing genetically modified immunoreactive cells) is administered, it will generally be in the form of a standard injectable unit dose (solution, suspension, emulsion).

[0253] Preparations include preparations for oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, buccal, sublingual, or suppository administration. In some embodiments, the agent or cell populations are administered parenterally. The term "parenteral" as used herein includes intravenous, intramuscular, subcutaneous, rectal, vaginal and intraperitoneal administration. In some embodiments, the agent or cell populations are administered to a subject using peripheral systemic delivery via intravenous, intraperitoneal, or subcutaneous injection.

[0254] In some embodiments, the compositions are provided as sterile liquid preparations, eg, isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which can, in some aspects, be adjusted to a selected pH with a buffer. Liquid formulations are generally easier to prepare than gels, other viscous formulations, and solid formulations. In addition, liquid formulations are slightly more convenient to administer, especially by injection. On the other hand, viscous compositions can be formulated in an appropriate viscosity range to provide longer periods of contact with particular tissues. Liquid or viscous compositions may contain carriers, which may be a solvent or dispersion medium containing, for example, water, saline, phosphate buffered saline, a polyol (eg, glycerol, propylene glycol, liquid polyethylene glycol), and appropriate mixtures thereof.

[0255] Sterile injectable solutions can be prepared by administering the cells in a solvent such as a mixture of an appropriate carrier, diluent or excipient such as sterile water, physiological saline, glucose, dextrose, or the like. The compositions may also be lyophilized. The compositions may contain adjuvants such as wetting, dispersing, or emulsifying agents (e.g., methylcellulose), pH buffering agents, gelling or viscosity enhancing agents, preservatives, flavoring agents, coloring agents, and the like, depending on the route of administration and the desired drug. In some respects, standard texts may be consulted for the preparation of appropriate preparations.

[0256] Various additives can be added that enhance the stability and sterility of the compositions, including antimicrobial preservatives, antioxidants, chelating agents, and buffers. Prevention of the action of microorganisms can be provided by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. Prolonged absorption of a pharmaceutical form for injection can be obtained using agents that delay absorption, for example, aluminum monostearate and gelatin.

[0257] Formulations for use in in vivo administration are generally sterile. Sterility can be easily obtained, for example, by filtration through sterile filter membranes.

[0258] For the prevention or treatment of a disease, the appropriate dose may depend on the type of disease to be treated, the type of agent or agents, the type of cell or recombinant receptor, the severity and course of the disease, whether the agent or cells are administered for preventive or therapeutic goals, from previous therapy, from the subject's medical history and response to the agent or cells, and from the judgment of the attending physician. The compositions, in some embodiments, are administered appropriately to a subject at one time or as a series of treatments.

[0259] In some cases, cell therapy is administered as a separate pharmaceutical composition containing cells. In some embodiments, a given dose is administered by administering a single bolus of cells or agents. In some embodiments, it is administered by multiple bolus of cells or agent, eg over a period of no more than 3 days, or by continuous infusion of cells or agent.

2. Time schedule for dosing and administration

[0260] In some embodiments, the implementation, the dose of cells is administered to subjects according to the proposed methods of combination therapy. In some embodiments, the size or timing of doses is determined depending on the particular disease or condition of the subject. You can empirically determine the size or timing of doses for a particular disease given the proposed description.

[0261] In certain embodiments, cells or distinct populations of cell subtypes are administered to a subject in the range of about 0.1 million to about 100 billion cells and/or the same number of cells per kilogram of the subject's body weight, e.g., about 0.1 million up to 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or range defined by any two of the above values), from 1 million to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or amounts in the range from to any two of the above values), e.g., about 10 million to about 100 billion cells (e.g., about 20 million cells, about 30 million cells, about 40 million cells, about 60 million cells, about 70 million cells, about 80 million cells, about 90 million cells, about 10 billion cells, about 25 billion cells, about 50 billion cells, about 75 billion cells, about 90 billion cells, or an amount in the range defined by any two of the above), and in some about 100 million cells to about 50 billion cells (e.g., about 120 million cells, about 250 million cells, about 350 million cells, about 450 million cells, about 650 million cells, about 800 million cells, about 900 million cells, about 3 billion cells, approximately 30 billion cells, approximately 45 billion cells) or any th amount in these ranges and/or per kilogram of the subject's body weight. Doses may vary depending on the specific parameters of the disease or disorder and/or the patient and/or other treatments. In some embodiments, such values refer to the number of cells expressing the recombinant receptor; in other embodiments, they refer to the number of T-lymphocytes or PBMCs, or to the cells administered, in general.

[0262] In some embodiments, the cell therapy comprises administering a dose containing a cell count of from or about 1 x 10 ⁵ to 1 x 10 ⁸ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, from or about 5 × 10 ⁵ to 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, or from or from about 1 x 10 ⁶ to 1 x 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, each value is inclusive. In some embodiments, cell therapy comprises administering a dose of cells containing a cell count of at least, or about, at least 1 x 10 ⁵ cells, in total, T-lymphocytes expressing recombinant receptors in total, or mononuclear cells of peripheral blood (PBMC), in general, for example, at least, or at least 1 × 10 ⁶ at least, or approximately, at least 1 × 10 ⁷ at least, or approximately, at least at least 1 × 10 ⁸ such cells.

[0263] In some embodiments, for example, when the subject is a human, the dose contains less than about 5 x 10 ⁸ cells expressing the recombinant receptor (e.g., CAR), T-lymphocytes, or peripheral blood mononuclear cells (PBMC), in total, for example, in the range from about 1 x 10 ⁶ to 5 x 10 ⁸ such cells, for example, 2 x 10 ⁶ , 5 x 10 ⁶ , 1 x 10 ⁷ , 5 x 10 ⁷ , 1 x 10 ⁸ , or 5 x 10 ⁸ such cells, in general, or the number in the range between any two of the above values.

[0264] In some embodiments, the number refers to the total number of CD3+ or CD8+, in some cases, also cells expressing the recombinant receptor (eg, CAR+). In some embodiments, the cell therapy comprises administering a dose containing from or about 1 x 10 ⁵ to 1 x 10 ⁸ CD3+ or CD8+ T lymphocytes in total, or CD3+ or CD8+ cells expressing the recombinant receptor from or about 5 x 10 ⁵ to 1 x 10 ⁷ CD3+ or CD8+ T lymphocytes in total, or CD3+ or CD8+ cells expressing the recombinant receptor, or from or about 1 x 10 ⁶ to 1 x 10 ⁷ CD3+ or CD8+ T lymphocytes , in general, or CD3+ or CD8+ cells expressing the recombinant receptor, each value inclusive. In some embodiments, the cell therapy comprises administering a dose containing a cell count of from or about 1 x 10 ⁵ to 1 x 10 ⁸ , in total, CD3+/CAR+ or CD8+/CAR+ cells from or about 5 x 10 ⁵ to 1 x 10 ⁷ total CD3+/CAR+ or CD8+/CAR+ cells, or from or about 1 x 10 ⁶ to 1 x 10 ⁷ total CD3+/CAR+ or CD8+/CAR+ cells, each inclusive.

[0265] In some embodiments, the dose of genetically engineered cells contains from or about 1 x 10 ⁵ to 5 x 10 ⁸ CAR-expressing T lymphocytes, in total, from 1 x 10 ⁵ to 2.5 x 10 ⁸ , CAR-expressing T-lymphocytes, in total 1 x 10 ⁵ to 1 x 10 ⁸ CAR-expressing T-lymphocytes, in total, 1 x 10 ⁵ to 5 x 10 ⁷ CAR-expressing T-lymphocytes, in total, 1 x 10 ⁵ to 2.5 x 10 ⁷ CAR-expressing T-lymphocytes, in total 1 x 10 ⁵ to 1 x 10 ⁷ CAR-expressing T-lymphocytes, in total 1 x 10 ⁵ to 5 x 10 ⁶ CAR-expressing T-lymphocytes, in total 1 × 10 ⁵ to 2.5 × 10 ⁶ CAR-expressing T-lymphocytes, in total 1 × 10 ⁵ to 1 × 10 ⁶ CAR-expressing T-lymphocytes 1 x 10 ⁶ to 5 x 10 ⁸ CAR-expressing T-lymphocytes in total, 1 x 10 ⁶ to 2.5 x 10 ⁸ CAR-expressing T-lymphocytes in total, 1 x 10 in total ⁶ to 1 x 10 ⁸ CAR-expressing T cells, overall 1 x 10 ⁶ to 5 x 10 ⁷ CAR-expressing T cells, overall 1 x 10 ⁶ to 2.5 x 10 ⁷ CAR-expressing T-lymphocytes, in total 1 × 10 ⁶ to 1 × 10 ⁷ CAR-expressing T-lymphocytes, in total 1 × 10 ⁶ to 5 × 10 ⁶ CAR-expressing T-lymphocytes, in overall 1 x 10 ⁶ to 2.5 x 10 ⁶ CAR-expressing T-lymphocytes, overall 2.5 x 10 ⁶ to 5 x 10 ⁸ CAR-expressing T-lymphocytes, overall 2.5 × 10 ⁶ to 2.5 × 10 ⁸ CAR-expressing T-lymphocytes, in total 2.5 × 10 ⁶ to 1 × 10 ⁸ CAR-expressing T-lymphocytes, in total 2.5 × 10 ⁶ to 5 x 10 ⁷ CAR-expressing T-lymphocytes, overall 2.5 x 10 ⁶ to 2.5 x 10 ⁷ CAR-expressing T-lymphocytes, overall 2.5 x 10 ⁶ to 1 x 10 ⁷ CAR-expressing T-lymphocytes, in total, from 2.5 × 10 ⁶ to 5 × 10 ⁶ CAR-expressing T-lymphocytes, in total, from 5 × 10 ⁶ to 5 × 10 ⁸ CAR-expressing T-lymphocytes, in overall 5 x 10 ⁶ to 2.5 x 10 ⁸ CAR-expressing T-lymphocytes, overall 5 x 10 ⁶ to 1 x 10 ⁸ CAR-expressing T-lymphocytes, overall 5 x 10 ⁶ to 5 × 10 ⁷ CAR-expressing T-lymphocytes, in overall 5 x 10 ⁶ to 2.5 x 10 ⁷ CAR-expressing T-lymphocytes, overall 5 x 10 ⁶ to 1 x 10 ⁷ CAR-expressing T-lymphocytes, overall 1 x 10 ⁷ to 5 x 10 ⁸ CAR-expressing T-lymphocytes, in total 1 x 10 ⁷ to 2.5 x 10 ⁸ CAR-expressing T-lymphocytes, in total 1 x 10 ⁷ to 1 x 10 ⁸ CAR-expressing T -lymphocytes, in total, from 1 × 10 ⁷ to 5 × 10 ⁷ CAR-expressing T-lymphocytes, in total, from 1 × 10 ⁷ to 2.5 × 10 ⁷ CAR-expressing T-lymphocytes, in total, from 2 .5 × 10 ⁷ to 5 × 10 ⁸ CAR-expressing T-lymphocytes, in total from 2.5 × 10 ⁷ to 2.5 × 10 ⁸ CAR-expressing T-lymphocytes, in total from 2.5 × 10 ⁷ to 1 x 10 ⁸ CAR-expressing T-lymphocytes, overall 2.5 x 10 ⁷ to 5 x 10 ⁷ CAR-expressing T-lymphocytes, overall 5 x 10 ⁷ to 5 x 10 ⁸ CAR- expressing T-lymphocytes, in total, from 5 × 10 ⁷ to 2.5 × 10 ⁸ CAR-expressing T-lymphocytes, in total, from 5 × 10 ⁷ to 1 × 10 ⁸ CAR-expressing T-lymphocytes, in total, from 1 × 10 ⁸ to 5 × 10 ⁸ CAR Express T-lymphocytes, in total, 1 x 10 ⁸ to 2.5 x 10 ⁸ CAR-expressing T-lymphocytes, in total, or 2.5 x 10 ⁸ to 5 x 10 ⁸ CAR-expressing T-lymphocytes, generally.

[0266] In some embodiments, the dose of genetically engineered cells contains at least, or at least about 1 x 10 ⁵ CAR-expressing cells, at least, or at least about 2.5 x 10 ⁵ CAR-expressing cells, at least, or at least about 5 × 10 ⁵ CAR-expressing cells, at least, or approximately, at least 1 × 10 ⁶ CAR-expressing cells, at least , or about at least 2.5 x 10 ⁶ CAR-expressing cells, at least, or about at least 5 x 10 ⁶ CAR-expressing cells, at least, or about, at least 1 x 10 ⁷ CAR expressing cells, at least or about at least 2.5 x 10 ⁷ CAR expressing cells, at least or about 5 x 10 ⁷ CAR expressing cells at least, or about, at least 1 x 10 ⁸ CAR-expressing cells, at least, or about, at least 2.5 x 10 ⁸ CA R-expressing cells, or at least or about at least 5×10 ⁸ CAR-expressing cells.

[0267] In some embodiments, cell therapy comprises administering a dose containing a cell count of from or about 1 x 10 ⁵ to 5 x 10 ⁸ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, from or about 5 × 10 ⁵ to 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, or from or from about 1 x 10 ⁶ to 1 x 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, each value is inclusive. In some embodiments, cell therapy comprises administering a cell dose containing a cell count of at least, or about, at least 1 x 10 ⁵ cells, in total, recombinant receptor-expressing T-lymphocytes in total, or mononuclear cells of peripheral blood (PBMC), in general, for example, at least, or at least 1 × 10 ⁶ at least, or about, at least 1 × 10 ⁷ at least, or approximately, at least at least 1 × 10 ⁸ such cells. In some embodiments, this number refers to the total number of CD3+ or CD8+, in some cases, also to cells expressing the recombinant receptor (eg, CAR+). In some embodiments, the cell therapy comprises administering a dose containing from or about 1 x 10 ⁵ to 5 x 10 ⁸ CD3+ or CD8+ T lymphocytes in total, or CD3+ or CD8+ cells expressing the recombinant receptor from or about 5 x 10 ⁵ to 1 x 10 ⁷ CD3+ or CD8+ T lymphocytes in total, or CD3+ or CD8+ cells expressing the recombinant receptor, or from or about 1 x 10 ⁶ to 1 x 10 ⁷ CD3+ or CD8+ T lymphocytes , in general, or CD3+ or CD8+ cells expressing the recombinant receptor, each value inclusive. In some embodiments, cell therapy comprises administering a dose containing a cell count of from or about 1 x 10 ⁵ to 5 x 10 ⁸ , in total, CD3+/CAR+ or CD8+/CAR+ cells from or about 5 x 10 ⁵ to 1 x 10 ⁷ cells, in total, CD3+/CAR+ or CD8+/CAR+, or from or about 1 x 10 ⁶ to 1 x 10 ⁷ cells of CD3+/CAR+ or CD8+/CAR+, in total, each value inclusive.

[0268] In some embodiments, the doses of T lymphocytes comprise CD4+ T lymphocytes, CD8+ or CD4+ T lymphocytes, and CD8+ T lymphocytes.

[0269] In some embodiments, for example, when the subject is human, the dose of CD8+ T lymphocytes, including the dose containing CD4+ and CD8+ T lymphocytes, contains between about 1 x 10 ⁶ and 5 x 10 ⁸ CD8+ cells expressing recombinant receptor (e.g., CAR), in general, e.g., in the range of about 5 x 10 ⁶ to 1 x 10 ⁸ such cells, e.g., 1 x 10 ⁷ , 2.5 x 10 ⁷ , 5 x 10 ⁷ , 7, 5 x 10 ⁷ , 1 x 10 ⁸ or 5 x 10 ⁸ such cells, in total, or between any two of the above values. In some embodiments, a plurality of doses are administered to a patient, and each of the doses, or the dose as a whole, may be within any of the values discussed above. In some embodiments, cell dosing comprises administering from or about 1 x 10 ⁷ to 0.75 x 10 ⁸ CD8+ T lymphocytes expressing the recombinant receptor, in total 1 x 10 ⁷ to 2.5 x 10 ⁷ T- recombinant CD8+ receptor-expressing lymphocytes, in total, from or about 1×10 ⁷ to 0.75×10 ⁸ recombinant CD8+ receptor-expressing T-lymphocytes, in total, each value inclusive. In some embodiments, cell dosing comprises administering 1 x 10 ⁷ , 2.5 x 10 ⁷ , 5 x 10 ⁷ 7.5 x 10 ⁷ , 1 x 10 ⁸ or 5 x 10 ⁸ or about 1 x 10 ⁷ , 2, 5 x 10 ⁷ , 5 x 10 ⁷ 7.5 x 10 ⁷ , 1 x 10 ⁸ or 5 x 10 ⁸ T-lymphocytes expressing the recombinant CD8+ receptor, in total.

[0270] In some embodiments, the dose of cells, e.g., T-lymphocytes expressing the recombinant receptor, is administered to the subject as a single dose, or it is administered only once within a period of two weeks, one month, three months, six months, 1 year or more.

[0271] In some embodiments, cell therapy comprises administering a dose containing a cell count that is at least, or at least about, or about 0.1 x 10 ⁶ cells/kg subject body weight, 0. 2 x 10 ⁶ cells/kg, 0.3 x 10 ⁶ cells/kg, 0.4 x 10 ⁶ cells/kg, 0.5 x 10 ⁶ cells/kg, 1 x 10 ⁶ cells/kg, 2.0 x 10 ⁶ cells/kg, 3 x 10 ⁶ cells/kg or 5 x 10 ⁶ cells/kg.

[0272] In some embodiments, cell therapy comprises administering a dose containing a cell count between or about 0.1×10 ⁶ cells/kg of the subject's body weight and 1.0×10 ⁷ cells/kg within between or approximately between 0.5 × 10 ⁶ cells/kg and 5 × 10 ⁶ cells/kg, within or between 0.5 × 10 ⁶ cells/kg and 3 × 10 ⁶ cells/kg, within between or approximately between 0.5 × 10 ⁶ cells/kg and 2 × 10 ⁶ cells/kg, within or between 0.5 × 10 ⁶ cells/kg and 1 × 10 ⁶ cells/kg, within between or about 1.0 x 10 ⁶ cells/kg body weight of the subject and 5 x 10 ⁶ cells/kg, between or about 1.0 x 10 ⁶ cells/kg and 3 x 10 ⁶ cells/kg, between or about 1.0 x 10 ⁶ cells/kg and 2 x 10 ⁶ cells/kg, between or about 2.0 x 10 ⁶ cells/kg of the subject's body weight and 5 x 10 ⁶ cells /kg, naho between or about 2.0 x 10 ⁶ cells/kg and 3 x 10 ⁶ cells/kg, or between or about 3.0 x 10 ⁶ cells/kg of the subject's body weight and 5 x 10 ⁶ cells/kg, each value inclusive.

[0273] In some embodiments, the dose of cells is between 2 x 10 ⁵ or about 2 x 10 ⁵ cells/kg and 2 x 10 ⁶ or about 2 x 10 ⁶ cells/kg, such as between 4 x 10 ⁵ or about 4 x 10 ⁵ cells/kg and 1 x 10 ⁶ or about 1 x 10 ⁶ cells/kg or between 6 x 10 ⁵ or about 6 x 10 ⁵ cells/kg and 8 x 10 ⁵ or about 8 x 10 ⁵ cells/kg. In some embodiments, the cell dose contains no more than 2 x 10 ⁵ cells (e.g., antigen-expressing, such as CAR-expressing cells) per kilogram of the subject's body weight (cells/kg), e.g., no more than (or about) 3 x 10 ⁵ cells/kg, no more (or approximately) 4 × 10 ⁵ cells/kg, no more than (or approximately) 5 × 10 ⁵ cells/kg, no more (or approximately) 6 × 10 ⁵ cells/kg, no more (or approximately) 7 × 10 ⁵ cells/kg, not more than (or approximately) 8 × 10 ⁵ cells/kg, not more than (or approximately) 9 × 10 ⁵ cells/kg, not more than (or approximately) 1 × 10 ⁶ cells/kg, or not more than (or approximately) 2 × 10 ⁶ cells/kg. In some embodiments, the cell dose contains at least, or at least about, 2 x 10 ⁵ cells (e.g., antigen-expressing cells, such as CAR-expressing cells) per kilogram of the subject's body weight (cells/kg) , for example, at least, or at least about 3 x 10 ⁵ cells/kg, at least, or at least about 4 x 10 ⁵ cells/kg, at least, or at least about 5 x 10 ⁵ cells/kg at least, or at least about 6 x 10 ⁵ cells/kg, or at least about 7 x 10 ⁵ cells/kg, at least or at least about 8 x 10 ⁵ cells/kg, or at least about 9 x 10 ⁵ cells/kg, or at least about 1 x 10 ⁶ cells /kg, or at least, or at least about 2×10 ⁶ cells/kg.

[0274] In the context of adoptive cell therapy, administering a given “dose” of cells encompasses administering a given number or amount of cells as a single composition and/or as a separate continuous administration, such as a single injection or continuous infusion, and also encompasses administering a given amount or the number of cells as a divided dose, offered in a variety of separate compositions or infusions, for a specified period of time, which does not exceed 3 days. Thus, in a certain context, a dose is a single or continuous administration of a given number of cells, delivered or started at one time. In some context, however, the dose is administered in multiple injections or infusions over a period of no more than three days, such as once a day for three days or two days, or as multiple infusions over a one-day period.

[0275] Thus, in some aspects, the dose cells are administered in a separate pharmaceutical composition. In some embodiments, the dose cells are administered in a plurality of compositions collectively containing the dose cells.

[0276] The term "split dose" refers to a dose that is divided in such a way that it is administered over several days. This type of dosing is covered by the present methods and is considered a separate dose. In some embodiments, the divided dose cells are administered in a plurality of dose cell compositions over a period of no more than three days.

[0277] Thus, the dose of cells can be administered as a divided dose. For example, in some embodiments, the dose may be administered to the subject over 2 days or over 3 days. Exemplary split dosing methods include administering 25% of the dose on the first day and administering the remaining 75% of the dose on the second day. In other embodiments, 33% of the dose may be administered on the first day and the remaining 67% administered on the second day. In some aspects, 10% of the dose is administered on the first day, 30% of the dose is administered on the second day, and 60% of the dose is administered on the third day. In some embodiments, the divided dose is not spread over more than 3 days.

[0278] In some embodiments, the implementation, the dose of cells, as a rule, is large enough to be effective in reducing and the severity of the disease.

[0279] In some embodiments, the cells are administered at the desired dose, which in some aspects contains the desired dose or number of cells or cell type(s) and/or the desired ratio of cell types. Thus, cell dosing in some embodiments refers to the total number of cells (or number per kg of body weight) and to the desired ratio of individual populations or subtypes, such as the ratio of CD4+ to CD8+. In some embodiments, cell dosing refers to the desired total number (or number per kg of body weight) of cells in particular populations or in particular cell types. In some embodiments, dosing refers to a combination of features such as the desired number of cells in general, the desired ratio, and the desired total number of cells in individual populations.

[0280] In some embodiments, populations or subtypes of cells, such as CD8+ and CD4+ T lymphocytes, are administered at a tolerable difference (or within) the desired dose of cells, in general, such as the desired dose of T lymphocytes. In some aspects, the desired dose is the desired number of cells, or the desired number of cells per unit body weight of the subject to which the cells are administered, eg, cells/kg. In some aspects, the desired dose is the minimum number (or higher) of cells or the minimum number of cells per unit body weight. In some aspects, from cells generally administered at the desired dose, distinct populations or subtypes are present at (or near) the desired output ratio (such as the ratio of CD4+ to CD8+), e.g., within a certain margin or error of such ratio.

[0281] In some embodiments, cells are administered at (or within) an acceptable desired dose difference for one or more distinct cell populations or subtypes, such as a desired CD4+ cell dose and/or a desired CD8+ cell dose. In some aspects, the desired dose is the desired number of cells of a given subtype or population, or the desired number of such cells per unit body weight of the subject to which the cells are administered, eg, cells/kg. In some aspects, the desired dose is the minimum number (or higher) of cells in a population or subtype, or the minimum number of cells in a population or subtype per unit body weight.

[0282] Thus, in some embodiments, dose refers to a desired fixed dose of cells in general and to a desired ratio, and/or refers to a desired fixed dose of one or more, e.g. each, individual subtypes or subpopulations. Thus, in some embodiments, dose refers to a desired fixed or minimum dose of T-lymphocytes and a desired ratio of CD4+ and CD8+ cells and/or refers to a desired fixed or minimum dose of CD4+ and/or CD8+ cells.

[0283] In some embodiments, cells are administered at (or within acceptable limits) the desired output ratio of multiple cell populations or subtypes, such as CD4+ and CD8+ cells or subtypes. In some aspects, the desired ratio may be a specific ratio, or may be a range of ratios, e.g., in some embodiments, the desired ratio (e.g., ratio of CD4+ to CD8+ cells) is between (or about) 1:5 and/or approximately) 5:1 (or greater than approximately 1:5 and less than approximately 5:1) or between (or approximately) 1:3 and (or approximately) 3:1 (or greater than approximately 1:3 and less than about 3:1), e.g., between (or about) 2:1 and (or about) 1:5 (or greater than about 1:5 and less than about 2:1, e.g., (or approximately) 5:1, 4.5:1, 4:1, 3.5:1, 3:1, 2.5:1, 2:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4:1, 1.3:1, 1.2:1, 1.1:1, 1:1, 1:1, 1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9: 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5 or 1:5 In some aspects, the allowable difference is in the range of about 1%, about 2% , about 3%, about 4% about 5%, about 10%, n about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50% of the desired ratio, including any value in these ranges.

[0284] In specific embodiments, the numbers and/or concentrations of cells refer to the number of cells expressing the recombinant receptor (eg, CAR). In other embodiments, cell numbers and/or concentrations refer to the number or concentration of all cells, T-lymphocytes, or peripheral blood mononuclear cells (PBMCs) administered.

[0285] In some aspects, the dose size is determined in relation to one or more criteria, such as the response of the subject to previous treatment, for example, chemotherapy, the severity of the subject's disease, for example, load, volume, tumor size or extent, spread or type of metastases, the stage and/or likelihood or frequency of development of toxic treatment outcomes in the subject, eg, CRS, macrogphage activation syndrome, tumor lysis syndrome, neurotoxicity, and/or host immune response against administered cells and/or recombinant receptors.

[0286] In some embodiments, administration of an immunomodulatory compound in combination with cells can significantly increase cell multiplication or proliferation and thus decrease the dose of cells that can be administered to a subject. In some cases, the present methods make it possible to administer a lower dose of such cells to achieve the same or better treatment efficacy as a dose in a method in which the cell therapy is administered without the administration of an immunomodulatory compound, for example, at least 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold or 10-fold less than the dose in a method in which cell therapy is administered without the administration of an immunomodulatory compound, for example, lenalidomide.

[0287] In some embodiments, the dose, for example, contains between or about between 5.0 x 10 ⁶ and 2.25 x 10 ⁷ , 5.0 x 10 ⁶ and 2.0 x 10 ⁷ , 5.0 × 10 ⁶ and 1.5 × 10 ⁷ , 5.0 × 10 ⁶ and 1.0 × 10 ⁷ , 5.0 × 10 ⁶ and 7.5 × 10 ⁶ , 7.5 × 10 ⁶ and 2.25 × 10 ⁷ , 7.5 × 10 ⁶ and 2.0 × 10 ⁷ , 7.5 × 10 ⁶ and 1.5 × 10 ⁷ , 7.5 × 10 ⁶ and 1.0 × 10 ⁷ , 1.0 × 10 ⁷ and 2.25 × 10 ⁷ , 1.0 × 10 ⁷ and 2.0 × 10 ⁷ , 1.0 × 10 ⁷ and 1.5 × 10 ⁷ , 1.5 × 10 ⁷ and 2.25 × 10 ⁷ , 1.5 × 10 ⁷ and 2.0 × 10 ⁷ , 2.0 × 10 ⁷ and 2.25 × 10 ⁷ . In some embodiments, the cell dose contains a number of cells that is between at least, or at least about 5 x 10 ⁶ , 6 x 10 ⁶ , 7 x 10 ⁶ , 8 x 10 ⁶ , 9 x 10 ⁶ , 10 x 10 ⁶ , and about 15 x 10 ⁶ recombinant receptor expressing cells, such as recombinant receptor expressing cells that are CD8+. In some embodiments, such a dose, for example, such a target number of cells, refers to cells expressing the recombinant receptor, in general, in the administered composition.

[0288] In some embodiments, for example, a lower dose contains less than about 5 x 10 ⁶ cells, cells expressing a recombinant receptor (e.g., CAR), T-lymphocytes, and/or PBMC per kilogram of the subject's body weight, for example, less than about 4.5 × 10 ⁶ , 4 × 10 ⁶ , 3.5 × 10 ⁶ , 3 × 10 ⁶ , 2.5 × 10 ⁶ , 2 × 10 ⁶ , 1.5 × 10 ⁶ , 1 × 10 ⁶ , 5 × 10 ⁵ , 2.5 x 10 ⁵ or 1 x 10 ⁵ such cells per kilogram of the subject's body weight. In some embodiments, the lower dose contains less than about 1 x 10 ⁵ , 2 x 10 ⁵ , 5 x 10 ⁵ , or 1 x 10 ⁶ such cells per kilogram of the subject's body weight, or has a value between any two of the values discussed above. In some embodiments, such values refer to numbers of cells expressing the recombinant receptor; in other embodiments, they refer to the number of T-lymphocytes or PBMCs, or to the injected cells in general.

[0289] In some embodiments, the subject receives multiple doses, such as two or more doses, or multiple consecutive doses of cells. In some embodiments, two doses are administered to the subject. In some embodiments, the subject receives a sequential dose, e.g., the second dose is administered approximately 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days after the first dose. In some embodiments, a plurality of consecutive doses are administered after the first dose such that an additional dose or doses are administered after the sequential dose is administered. In some aspects, the number of cells administered to a subject at an additional dose is the same or similar to the first dose and/or subsequent dose. In some embodiments, the additional dose or doses are greater than previous doses. In some embodiments, one or more consecutive doses of cells may be administered to a subject. In some embodiments, the sequential dose of cells is administered more or more than about 7 days, 14 days, 21 days, 28 days, or 35 days after the start of the first dose of cells. The sequential dose of cells may be greater than, about the same as, or less than the first dose. In some embodiments, administration of T cell therapy, such as administration of the first and/or second dose of cells, may be repeated.

[0290] In some embodiments, the initiation of administration of a cell therapy, e.g., a dose of cells or the first dose of a divided dose of cells, is before (before), simultaneously with, or after (sequentially or serially) administration of an immunomodulatory compound, for example, lenalidomide.

[0291] In some embodiments, the cell dose, or sequential cell dose, is administered concurrently with the initiation of administration of the immunomodulatory compound in accordance with combination therapy methods. In some embodiments, the cell dose, or sequential cell dose, is administered on the same day that the administration of the immunomodulatory compound is started according to the combination therapy methods. In some embodiments, a dose of cells, or a sequential dose of cells, is administered within 1 day, within 2 days, within 3 days, within 4 days, within 5 days, within 6 days, or within 7 days of onset administering an immunomodulatory compound according to combination therapy methods.

[0292] In some embodiments, a dose of cells or a sequential dose of cells is administered prior to the start or start of administration of the immunomodulatory compound in accordance with the proposed combination therapy. In some embodiments, the cell dose is administered at least, or at least about 1 hour, at least, or at least about 2 hours, at least, or at least about after 3 hours, at least, or at least about 6 hours, at least, or at least about 12 hours, at least, or at least about 1 day, by at least, or at least about 2 days later, at least, or at least about 3 days later, at least, or at least about 4 days later, at least, or, at least about 5 days, at least, or at least about 6 days, at least, or at least about 7 days, at least, or at least about after 12 days, at least, or at least about 14 days, at least, or at least about 15 days, at least at least, or at least after about 21 days, at least, or at least after about 28 days, at least, or at least after about 30 days, at least, or, at least about 35 days, at least, or at least about 42 days, at least, or at least about 60 days, or at least, or at least about 90 days prior to the administration of an immunomodulatory compound in accordance with the proposed combination therapy.

[0293] In some embodiments, administration of an immunomodulatory compound (e.g., lenalidomide) according to the proposed combination therapy occurs at a time point at which previous administration of immunotherapy (e.g., T cell therapy, such as CAR T cell therapy) is, or likely to be, related to a decrease in T-lymphocyte functionality relative to T-lymphocyte functionality at the time immediately before the start of immunotherapy (eg, T-cell therapy, such as CAR T-lymphocyte therapy) or at a previous time point after initiation of T-cell therapy. In some embodiments, the method comprises, after administration of a dose of cells from a T cell therapy, such as adoptive T cell therapy, but prior to administration of an immunomodulatory compound, evaluating a sample from the subject for one or more T cell functions, such as proliferation or persistence. cells, for example, as determined by their level or amount in the blood, or other phenotypes or desired treatment outcomes, as described herein, for example, as those described in Section III. Various parameters for determining or evaluating a combination therapy regimen are described in Section III.

B. Administration of an Immunomodulatory Compound

[0294] Proposed combination therapy methods, compositions, combinations, kits, and uses include administering an immunomodulatory compound, such as a structural or functional analog or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor, such as lenalidomide, which can be administered before, after, during, simultaneously or almost simultaneously, sequentially and/or alternately with the introduction of T-cell therapy, for example, with the introduction of T-lymphocytes expressing the chimeric antigen receptor (CAR).

[0295] In some embodiments, the immunomodulatory compound is a compound from the class of immunomodulatory compounds that is a structural or functional analog or derivative of thalidomide and/or an E3 ubiquitin ligase inhibitor.

[0296] In some embodiments, the immunomodulatory compound binds to cereblon (CRBN). In some embodiments, the immunomodulatory compound binds to the CRBN and E3 ubiquitin ligase complex. In some embodiments, the immunomodulatory compound binds to CRBN and to the CRBN-E3 ubiquitin ligase complex. In some embodiments, the immunomodulatory compound up-regulates CRBN protein or gene expression. In some aspects, CRBN is a substrate adapter for CRL4CRBN and E3 ubiquitin ligase and modulates enzyme specificity. In some embodiments, binding to CRB or to the complex of CRBN and E3 ubiquitin ligase inhibits the activity of E3 ubiquitin ligase. In some embodiments, the immunomodulatory compound induces ubiquitination of KZF1 (Ikaros) and IKZF3 (Aiolos) and/or induces degradation of IKZF1 (Ikaros) and IKZF3 (Aiolos). In some embodiments, the immunomodulatory compound induces ubiquitination of casein kinase 1A1 (CK1α) by CRL4CRBN and E3 ubiquitin ligase. In some embodiments, ubiquitination of CK1α results in degradation of CK1α.

[0297] In some embodiments, the immunomodulatory compound is an inhibitor of the Ikaros transcription factor (IKZF1). In some embodiments, the immunomodulatory compound enhances Ikaros ubiquitination. In some embodiments, the immunomodulatory compound enhances the degradation of Ikaros. In some embodiments, the immunomodulatory compound down-regulates the expression of an Ikaros protein or gene. In some embodiments, administration of an immunomodulatory compound causes a decrease in the level of Ikaros proteins.

[0298] In some embodiments, the immunomodulatory compound is an inhibitor of the Aiolos transcription factor (IKZF3). In some embodiments, the immunomodulatory compound enhances Aiolos ubiquitination. In some embodiments, the immunomodulatory compound enhances the degradation of Aiolos. In some embodiments, the immunomodulatory compound down-regulates the expression of an Aiolos protein or gene. In some embodiments, administration of an immunomodulatory compound causes a decrease in the level of the Aiolos protein.

[0299] In some embodiments, the immunomodulatory compound is an inhibitor of both Ikaros (IKZF1) and Aiolos (IKZF3) transcription factors. In some embodiments, the immunomodulatory compound enhances ubiquitination of both Ikaros and Aiolos. In some embodiments, the immunomodulatory compound enhances the degradation of both Ikaros and Aiolos. In some embodiments, the immunomodulatory compound enhances ubiquitination and degradation of both Ikaros and Aiolos. In some embodiments, administration of an immunomodulatory compound causes a decrease in the levels of both the Aiolos proteins and the levels of the Ikaros proteins.

[0300] In some embodiments, the immunomodulatory compound is a selective cytokine inhibitory drug (SelCID). In some embodiments, the immunomodulatory compound inhibits phosphodiesterase-4 (PDE4) activity. In some embodiments, the immunomodulatory compound inhibits the enzymatic activity of CDC25 phosphatases. In some embodiments, the immunomodulatory compound alters the intracellular targeted migration of CDC25 phosphatases.

[0301] In some embodiments, the immunomodulatory compound in combination therapy is thalidomide (2-(2,6-dioxopiperidin-3-yl)-1H-isoindol-1,3(2H)-dione) or an analog or derivative of thalidomide. In certain embodiments, the thalidomide derivative includes structural variants of thalidomide that have similar biological activity. Exemplary thalidomide derivatives include, but are not limited to, lenalidomide (REVLIMMUNOMODULATORY COMPOUND™; Celgene Corporation), pomalidomide (also known as ACTIMMUNOMODULATORY COMPOUND™ or POMALYST™ (Celgene Corporation)), CC-1088, CDC-501, and CDC-801 , and the compounds described in US patent No. 5712291; 7320991; and 8716315; in US Patent Application No. 2016/0313300; and PCT Publication Nos. WO 2002/068414 and WO 2008/154252.

[0302] In some embodiments, the immunomodulatory compound is 1-oxo- and 1,3-dioxo-2-(2,6-dioxo-piperidin-3-yl)isoindolines substituted with amino on the benzo ring, as described in the US patent No. 5635517, which is incorporated herein by reference.

[0303] In some embodiments, the immunomodulatory compound is a compound of the following formula:

where one of X and Y is -C(O)- and the other of X and Y is -C(O)- or -CH ₂ - and R ⁵ is hydrogen or lower alkyl or a pharmaceutically acceptable salt thereof . In some embodiments, X is -C(O)- and Y is -CH ₂ -. In some embodiments, both X and Y are -C(O)-. In some embodiments, R ⁵ is hydrogen. In other embodiments, R ⁵ is methyl.

[0304] In some embodiments, the immunomodulatory compound is a compound that belongs to the class of substituted 2-(2,6-dioxpiperidin-3-yl)phthalimmunomodulatory compounds and substituted 2-(2,6-dioxpiperidin-3-yl)- 1-oxoisoindoles such as those described in US Pat. Nos. 6,281,230; 6316471; 6,335,349 and 6,476,052 and International Patent Application No. PCT/US97/13375 (International Publication No. WO 98/03502), each of which is incorporated herein by reference.

[0305] In some embodiments, the immunomodulatory compound is a compound of the following formula:

where

one of X and Y is -C(O)- and the other of X and Y is -C(O)- or -CH ₂ -;

(1) each of R^one, R², R³ and R^four independently represents a halogen, alkyl with 1-4 carbon atoms or alkoxy from 1-4 carbon atoms, or

(2) one of R^one, R³, R^four and R⁵represents -NHR^aand the rest R^one, R², R³ and R^fourrepresent hydrogen, where R^arepresents hydrogen or alkyl with 1-8 carbon atoms;

R ⁵ is hydrogen or alkyl with 1-8 carbon atoms, benzyl or halogen;

with the proviso that R ⁵ is not hydrogen, if X and Y are -C(O)- and (i) each of R ¹ , R ² , R ³ and R ⁴ is fluoro or (ii) one of R ¹ , R ² , R ³ and R ⁴ is amino;

or a pharmaceutically acceptable salt thereof.

[0306] In some embodiments, the immunomodulatory compound is a compound that belongs to the class of isoindole-immunomodulatory compounds described in US Patent No. 7,091,353, US Patent Publication No. 2003/0045552, and International Application No. publication No. WO02/059106), each of which is incorporated herein by reference. For example, in some embodiments, the immunomodulatory compound is [2-(2,6-dioxo-piperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-ylmethyl]amide; (2-(2,6-dioxo-piperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-ylmethyl)-carbamic acid tert-butyl ester; 4-(aminomethyl)-2-(2,6-dioxo(3-piperidyl))isoindoline-1,3-dione; N-(2-(2,6-dioxo-piperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-4-ylmethyl)acetamide; N-{(2-(2,6-dioxo(3-piperidyl)-1,3-dioxoisoindolin-4-yl)methyl}cyclopropylcarboxamide; 2-chloro-N-{(2-(2,6-dioxo (3-piperidyl))-1,3-dioxoisoindolin-4-yl)methyl}acetamide; N-(2-(2,6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl)- 3-pyridylcarboxamide;3-{1-oxo-4-(benzylamino)isoindolin-2-yl}piperidin-2,6-dione;2-(2,6-dioxo(3-piperidyl))-4-(benzylamino) isoindolin-1,3-dione; N-{(2-(2,6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl)methyl}propanamide; N-{(2-(2, 6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl)methyl}-3-pyridylcarboxamide; N-{(2-(2,6-dioxo(3-piperidyl))-1,3- dioxoisoindolin-4-yl)methyl}heptanamide;N-{(2-(2,6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl)methyl}-2-furylcarboxamide;{N-( 2-(2,6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl)carbamoyl}methyl acetate;N-(2-(2,6-dioxo(3-piperidyl))-1,3 -dioxoisoindolin-4-yl)pentanamide;N-(2-(2,6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl)-2-thienylcarboxamide;N-{[2-(2 ,6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl]methyl}-(butylam ino)carboxamide; N-{[2-(2,6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl]methyl}(octylamino)carboxamide; or N-{[2-(2,6-dioxo(3-piperidyl))-1,3-dioxoisoindolin-4-yl]methyl}(benzylamino)carboxamide.

[0307] In some embodiments, the immunomodulatory compound is a compound that belongs to the class of isoindole-immunomodulatory compounds described in U.S. Patent Application Publication No. 2002/0045643, International Publication No. WO 98/54170, and U.S. Patent No. 6,395,754, each of which is incorporated herein by reference. In some embodiments, the immunomodulatory compound is the tetrasubstituted 2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolines described in US Pat. No. 5,798,368, which is incorporated herein by reference. In some embodiments, the immunomodulatory compound is 1-oxo and 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)isoindolines as described in US Pat. No. 6,403,613, which is incorporated herein by reference. In some embodiments, the immunomodulatory compound is 1-oxo or 1,3-dioxoisoindoline substituted at the 4- or 5-position of the indoline ring as described in US Pat. No. 6,380,239 and US Pat. No. 7,244,759, both of which are incorporated herein. as links.

[0308] In some embodiments, the immunomodulatory compound is 2-(4-amino-1-oxo-1,3-dihydro-isoindol-2-yl)-4-carbamoyl-butyric acid or 4-(4-amino- 1-oxo-1,3-dihydro-isoindol-2-yl)-4-carbamoylbutyric acid. In some embodiments, the immunomodulatory compound is 4-carbamoyl-4-{4-[(furan-2-yl-methyl)amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}butyric acid, 4-carbamoyl-2-{4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}-butyric acid, 2-{ 4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}-4-phenylcarbamoyl-butyric acid or 2-{4-[(furan -2-yl-methyl)amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}glutaric acid.

[0309] In some embodiments, the immunomodulatory compound is isoindolin-1-one or isoindolin-1,3-dione substituted at the 2-position with 2,6-dioxo-3-hydroxypiperidin-5-yl, as described in the US patent No. 6458810, which is incorporated herein by reference. In some embodiments, the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione, or an enantiomer or a mixture of enantiomers thereof; or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In some embodiments, the immunomodulatory compound is 3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidin-2,6-dione .

[0310] In some embodiments, the immunomodulatory compound is as described in Oshima, K. et al., Nihon Rinsho., 72(6):1130-5 (2014); Millrine, D. et al., Trends Mol Med., 23(4):348-364 (2017); and Collins, et al., Biochem J., 474(7):1127-1147 (2017).

[0311] In some embodiments, the immunomodulatory compound is an E3 ubiquitin ligase inhibitor. In some embodiments, the immunomodulatory compound is a thalidomide derivative. In some embodiments, the immunomodulatory compound is a structural and/or functional analogue of thalidomide. In some embodiments, the immunomodulatory compound is lenalidomide, pomalidomide, avadomide, or a pharmaceutically acceptable salt thereof.

[0312] In some embodiments, the immunomodulatory compound is lenalidomide, pomalidomide, avadomide, a stereoisomer of lenalidomide, pomalidomide, avadomide, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof. In some embodiments, the immunomodulatory compound is lenalidomide, a stereoisomer of lenalidomide, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, clathrate, or polymorph thereof.

(Формула I), или представляет собой их энантиомер или смесь энантиомеров; или их фармацевтически приемлемую соль, сольват, гидрат, сокристалл, клатрат или полиморф (ниже Соединение 1).[0313] In some embodiments, the immunomodulatory compound is avadomide, which is also known as 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione, having the following structure

(Formula I), or represents their enantiomer or mixture of enantiomers; or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof (below Compound 1).

[0314] In some embodiments, the immunomodulatory compound is an enantiomer or mixture of enantiomers of 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione, or their a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph of 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione. In some embodiments, the immunomodulatory compound is a 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione solvate. In some embodiments, the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione hydrate. In some embodiments, the immunomodulatory compound is a pharmaceutically acceptable salt of 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione. In some embodiments, the immunomodulatory compound is a 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione polymorph. In some embodiments, the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione. In some embodiments, the immunomodulatory compound has the structure of Formula I.

[0315] In some embodiments, the immunomodulatory compound is lenalidomide, also known as 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione , or is an enantiomer or a mixture of their enantiomers; or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In some embodiments, lenalidomide is 2,6-piperidinedione, 3-(4-amino-1,3-dihydro-1-oxo-2H-isoindol-2-yl)-, 3-(4-amino-1- oxo-1,3-dihydro-2H-isoindol-2-yl)-2,6-piperidinedione, 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)-2 ,6-piperidinedione, 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione, 3-(4-amino-1-oxo-1 ,3-dihydro-2H-isoindol-2-yl)piperidin-2,6-dione, 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-2, 6-dione, all of which can be used interchangeably, or is an enantiomer or a mixture of their enantiomers; or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

[0316] In some embodiments, the immunomodulatory compound is (R)-3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione. In some embodiments, the immunomodulatory compound is (S)-3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione. In some embodiments, the immunomodulatory compound is a mixture of (R)-3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione and (S) -3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione.

(Формула II), или энантиомер или смесь энантиомеров; или их фармацевтически приемлемую соль, сольват, гидрат, сокристалл, клатрат, или полиморф. В некоторых вариантах осуществления, иммуномодуляторное соединение представляет собой

(Формула IIA), или его фармацевтически приемлемую соль, сольват, гидрат, сокристалл, клатрат или полиморф. В других вариантах осуществления, иммуномодуляторное соединение представляет собой

(Формула IIB), или его фармацевтически приемлемую соль, сольват, гидрат, сокристалл, клатрат или полиморф. В определенных вариантах осуществления, иммуномодуляторное соединение содержит смесь

(Формула IIA) и

(Формула IIB), или их фармацевтически приемлемые соли, сольваты, гидраты, сокристаллы, клатраты или полиморфы. [0317] In some embodiments, the immunomodulatory compound is

(Formula II), or an enantiomer or a mixture of enantiomers; or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof. In some embodiments, the immunomodulatory compound is

(Formula IIA), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In other embodiments, the immunomodulatory compound is

(Formula IIB), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In certain embodiments, the immunomodulatory compound comprises a mixture

(Formula IIA) and

(Formula IIB), or their pharmaceutically acceptable salts, solvates, hydrates, co-crystals, clathrates or polymorphs.

[0318] In some embodiments, the immunomodulatory compound is an enantiomer or mixture of enantiomers of 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph of 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione. In some embodiments, the immunomodulatory compound is a solvate of (R)-3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione and/or ( S)-3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione. In some embodiments, the immunomodulatory compound is (RS)-3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione hydrate and/or ( S)-3-(4-Amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione. In some embodiments, the immunomodulatory compound is a pharmaceutically acceptable salt of (R)-3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione and/ or (S)-3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione. In some embodiments, the immunomodulatory compound is lenalidomide or 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione. In some embodiments, the immunomodulatory compound has the structure of Formula II. In some embodiments, the immunomodulatory compound has the structure of Formula IIA or Formula IIB, or is a mixture thereof.

(Формула III) или энантиомер или смесь их энантиомеров; или их фармацевтически приемлемую соль, сольват, гидрат, сокристалл, клатрат или полиморф. В некоторых вариантах осуществления, иммуномодуляторное соединение представляет собой

(Формула IIIA), или его фармацевтически приемлемую соль, сольват, гидрат, сокристалл, клатрат или полиморф. В других вариантах осуществления, иммуномодуляторное соединение представляет собой

(Формула IIIB) или его фармацевтически приемлемую соль, сольват, гидрат, сокристалл, клатрат или полиморф. В определенных вариантах осуществления, иммуномодуляторное соединение содержит смесь

(Формула IIIA) и

(Формула IIIB), или их фармацевтически приемлемые соли, сольваты, гидраты, сокристаллы, клатраты или полиморфы. [0319] In some embodiments, the immunomodulatory compound is pomalidomide, which is also known as 4-amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione, or is an enantiomer or a mixture thereof enantiomers; or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In some embodiments, the immunomodulatory compound is

(Formula III) or an enantiomer or a mixture of their enantiomers; or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In some embodiments, the immunomodulatory compound is

(Formula IIIA), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In other embodiments, the immunomodulatory compound is

(Formula IIIB) or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof. In certain embodiments, the immunomodulatory compound comprises a mixture

(Formula IIIA) and

(Formula IIIB), or their pharmaceutically acceptable salts, solvates, hydrates, co-crystals, clathrates or polymorphs.

[0320] In some embodiments, the immunomodulatory compound is an enantiomer or mixture of enantiomers of 4-amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph of 4-amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione. In some embodiments, the immunomodulatory compound is (R)-4-amino-2-(2,6-dioxopiperidin-3-yl)-isoindoline-1,3-dione and/or (S)-4-amino-2 -(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof (R)-4-amino-2-(2,6-dioxpiperidine -3-yl)isoindoline-1,3-dione and/or (S)-4-amino-2-(2,6-diox-piperidin-3-yl)isoindoline-1,3-dione. In some embodiments, the immunomodulatory compound is a solvate of (R)-4-amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione and/or (S)-4-amino-2 -(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione. In some embodiments, the immunomodulatory compound is a hydrate of (R)-4-amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione and/or (S)-4-amino-2 -(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione. In some embodiments, the immunomodulatory compound is a pharmaceutically acceptable salt of (R)-4-amino-2-(2,6-diox-piperidin-3-yl)isoindoline-1,3-dione and/or (S)-4-amino -2-(2,6-diox-piperidin-3-yl)isoindoline-1,3-dione. In some embodiments, the immunomodulatory compound is (R)-4-amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione, (S)-4-amino-2-(2 ,6-dioxopiperidin-3-yl)isoindoline-1,3-dione or a mixture thereof in any ratio. In some embodiments, the immunomodulatory compound has the structure of Formula III. In some embodiments, the immunomodulatory compound has the structure of Formula IIIA or Formula IIIB, or a mixture thereof.

[0321] In some embodiments, the immunomodulatory compound is or contains lenalidomide. Lenalidomide is FDA approved for the treatment of multiple myeloma, 5q deletion myelodysplastic syndrome, and most commonly recurrent/refractory mantle cell lymphoma (MCL). Lenalidomide is a synthetic derivative of thalidomide and is currently understood to have a variety of immunomodulatory effects, including increased immune synapse formation between T lymphocyte and antigen presenting cells (APCs). For example, in some cases, lenalidomide modulates the responses and outcomes of T-lymphocytes with increased production of interleukin (IL)-2 in CD4+ and CD8+ T-lymphocytes, induces a shift in T-helper (Th) responses from Th2 to Th1, inhibits the expansion of the T regulatory subset -lymphocytes (Treg), and improves the functioning of immunological synapses in follicular lymphoma (FL) and chronic lymphocytic leukemia (CLL) (Otahal et al., Oncoimmunology (2016) 5(4):e1115940). Lenalidomide also has direct tumoricidal activity in patients with multiple myeloma (MM) and directly and indirectly modulates CLL tumor cell survival through the influence of supporting cells such as stromal cells found in the microenvironment of lymphoid tissues.

1. Compositions and preparations

[0322] In some embodiments of the combination therapy methods, compositions, combinations, kits, and uses provided herein, the combination therapy may be administered in one or more compositions, such as a pharmaceutical composition containing an immunomodulatory compound, such as lenalidomide.

[0323] In some embodiments, a composition, e.g., a pharmaceutical composition, containing an immunomodulatory compound, e.g., lenalidomide, may include carriers, such as a diluent, excipient, excipient, or vehicle, with which the immunomodulatory compound, e.g., lenalidomide, is administered. and/or cells. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E. W. Martin. Such compositions will contain a therapeutically effective amount of an immunomodulatory compound, eg lenalidomide, generally in purified form, together with an appropriate amount of carrier, to provide a form for proper administration to the patient. Such pharmaceutical carriers may be sterile liquids such as water and oils, including petroleum, animal, vegetable or synthetic oils such as peanut oil, soybean oil, mineral oil and sesame oil. Saline solutions and aqueous solutions of dextrose and glycerol can also be used as liquid carriers, in particular for injectable solutions. Pharmaceutical compositions may contain any one or more of diluents, excipients, release agents, binders, coatings, fillers, flavoring agents, coloring agents, lubricants, glidants, preservatives, detergents, sorbents, emulsifying agents, pharmaceutical excipients, pH buffering agents, or sweeteners and their combinations. In some embodiments, the pharmaceutical composition may be a liquid, a solid, a lyophilized powder, in the form of a gel, and/or a combination thereof. In some aspects, the choice of carrier is determined in part by the particular inhibitor and/or route of administration.

[0324] Pharmaceutically acceptable carriers are generally non-toxic to patients at the doses and concentrations used and include, but are not limited to: buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes (eg Zn-protein complexes) and/or non-ionic surfactants such as polyethylene glycol (PEG), stabilizers and/or preservatives. Compositions containing an immunomodulatory compound, such as lenalidomide, may also be lyophilized.

[0325] In some embodiments, pharmaceutical compositions can be formulated for administration by any known route, including intramuscular, intravenous, intradermal, intralesional, intraperitoneal injection, subcutaneous, intratumoral, epidural, nasal, oral, vaginal, rectal, topical , external, intra-aural, inhalation, buccal (eg, sublingual) and transdermal administration, or any other method. In some embodiments, other routes of administration are also contemplated. In some embodiments, administration is by bolus infusion, by injection, such as intravenous or subcutaneous injection, intraocular injection, periocular injection, subretinal injection, intravitreal injection, transseptal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjunctival injection, subconjunctival injection , Tenon's capsule injection, retrobulbar injection, peribulbar injection, or posterior juxtascleral delivery. In some embodiments, administration is via parenteral, intrapulmonary, and intranasal, and, if desired, topical treatments administered within the affected tissues. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. In some embodiments, the prescribed dose is administered via a single bolus injection. In some embodiments, it is administered via multiple bolus administrations, such as over a period of no more than 3 days, or via continuous infusion administration.

[0326] In some embodiments, the implementation, the introduction can be local, external or systemic, depending on the locus of treatment. In some embodiments, local administration to the area in need of treatment can be achieved, for example, but not limited to, by local infusion during surgery, external application, for example, in combination with a wound dressing after surgery, by injection, via a catheter, via a suppository, or via an implant. In some embodiments, the compositions can also be administered together with other biologically active agents, either sequentially, alternately, or in the same composition. In some embodiments, administration may also include controlled release systems, including controlled release formulations and controlled release devices, such as by means of a pump. In some embodiments, the administration is oral.

[0327] In some embodiments, the pharmaceutically and therapeutically active compounds and derivatives thereof are typically formulated and administered in single unit dosage forms or in multiple unit dosage forms. Each unit dose contains a predetermined amount of a therapeutically active compound sufficient to produce the desired therapeutic effect in association with the necessary pharmaceutical carrier, vehicle or diluent. In some embodiments, unit dosage units include, but are not limited to, tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, and oral solutions or suspensions, and oil-in-water emulsions containing appropriate amounts of the compounds or their pharmaceutically acceptable derivatives. Unit dosage units may be contained in ampoules and syringes or in individually packaged tablets or capsules. Single unit doses may be administered in portions or in multiples. In some embodiments, the plurality of unit dose units is a plurality of identical unit dose units packaged in a single administration container in the form of divided unit dose units. Examples of multiple unit dosage units include vials, tablet or capsule bottles, or pint or gallon bottles.

[0328] Active ingredients can be packaged in microcapsules, colloidal drug delivery systems (eg, liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules), or macroemulsions. In certain embodiments, a pharmaceutical composition containing an immunomodulatory compound, such as lenalidomide, is formulated as an inclusion complex, such as a cyclodextrin inclusion complex, or as a liposome. Liposomes can serve to target host cells (eg, T-lymphocytes or NK cells) to a specific tissue. Many methods are available for preparing liposomes, such as those described, for example, in Szoka et al., Ann. Rev. Biophys. Bioeng., 9: 467 (1980), and US Pat. Nos. 4,235,871, 4,501,728, 4,837,028 and 5,019,369.

[0329] A pharmaceutical composition containing an immunomodulatory compound, such as lenalidomide, in some aspects may use timed release, sustained release, and timed release delivery systems such that delivery of the composition occurs prior to sensitization of the site to be treated, and enough time before that to cause sensitization. Many types of delivery and release systems are available and known. Such systems can eliminate multiple administrations of the composition, thereby increasing convenience for the subject and the clinician.

[0330] Compositions containing an immunomodulatory compound, such as lenalidomide, may also be lyophilized. The compositions may contain adjuvants such as wetting, dispersing, or emulsifying agents (e.g., methylcellulose), pH buffering agents, gelling or viscosity-increasing additives, preservatives, flavoring agents, coloring agents, and the like, depending on the route of administration and the desired drug. In some respects, standard texts may be consulted for the preparation of appropriate preparations.

[0331] Various additives can be added that enhance the stability and sterility of the compositions, including antimicrobial preservatives, antioxidants, chelating agents, and buffers. Prevention of the action of microorganisms can be provided by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. Prolonged absorption of the pharmaceutical form for injection can be obtained using agents that delay absorption, for example, aluminum monostearate and gelatin.

[0332] Sustained release formulations can be prepared. Suitable examples of sustained release formulations include semi-permeable matrices of solid hydrophobic polymers containing the antibody, these matrices are in the form of molded articles such as films or microcapsules.

[0333] In some embodiments, the composition containing the immunomodulatory compound, eg, lenalidomide, is administered in the form of a salt, eg, a pharmaceutically acceptable salt. Suitable pharmaceutically acceptable acid addition salts include salts derived from mineral acids such as hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric and sulfuric acids, and from organic acids such as tartaric, acetic, citric, malic, lactic, fumaric, benzoic, glycolic, gluconic, succinic and arylsulfonic acid, for example p-toluenesulfonic acid.

2. Timing Schedule for Dosing an Immunomodulatory Compound

[0334] In some embodiments, the proposed method of combination therapy includes administering to the subject a therapeutically effective amount of an immunomodulatory drug (immunomodulatory compound), for example, lenalidomide, and a cellular therapy, such as T-cell therapy (for example, CAR-expressing T-lymphocytes) .

[0335] In some embodiments, the administration of an immunomodulatory compound, for example, lenalidomide, begins before, after, during, during the course, simultaneously or almost simultaneously, sequentially and/or alternately with the introduction of cell therapy, such as T-cell therapy (for example, CAR-expressing T-lymphocytes). In some embodiments, the method includes initiating administration of an immunomodulatory compound, such as lenalidomide, prior to administration of T cell therapy. In other embodiments, the method includes initiating administration of an immunomodulatory compound, such as lenalidomide, following administration of T cell therapy. In some embodiments, the dosing timing includes initiating administration of an immunomodulatory compound, such as lenalidomide, at the same time or simultaneously with the administration of T cell therapy.

[0336] In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered in a cycle. In some embodiments, the cycle includes an administration period in which an immunomodulatory compound, eg, lenalidomide, is administered followed by a rest period during which an immunomodulatory compound, eg, lenalidomide, is not administered. In some embodiments, the total number of cycle days, e.g., from start of immunomodulatory compound administration, is greater than or about greater than, or about 21 days, 28 days, 30 days, 40 days, 50 days, 60 days or more.

[0337] In some embodiments, the start of administration of an immunomodulatory compound, for example, lenalidomide, is carried out in at least one cycle, and the start of administration of T-cell therapy occurs on the same day, optionally simultaneously. In some embodiments, the initiation of administration of an immunomodulatory compound, eg, lenalidomide, in at least one cycle occurs prior to the initiation of administration of T cell therapy. In some embodiments, the initiation of administration of an immunomodulatory compound, eg, lenalidomide, in at least one cycle occurs at the same time or on the same day as the initiation of administration of T cell therapy. In some embodiments, the immunomodulatory compound, e.g., lenalidomide, is administered from or about 0 to 30 days, e.g., 0 to 15 days, 0 to 6 days, 0 to 96 hours, 0 to 24 hours, 0 to 12 hours, 0 to 6 hours or 0 to 2 hours, 2 hours to 15 days, 2 hours to 6 days, 2 hours to 96 hours, 2 hours to 24 hours, 2 hours to 12 hours , 2 hours to 6 hours, 6 hours to 30 days, 6 hours to 15 days, 6 hours to 6 days, 6 hours to 96 hours, 6 hours to 24 hours, 6 hours to 12 hours, 12 hours to 30 days 12 hours to 15 days 12 hours to 6 days 12 hours to 96 hours 12 hours to 24 hours 24 hours to 30 days 24 hours to 15 days 24 hours to 6 days, 24 hours to 96 hours, 96 hours to 30 days, 96 hours to 15 days, 96 hours to 6 days, 6 days to 30 days, 6 days to 15 days, or 15 days days to 30 days before starting T-cell therapy. In some aspects, an immunomodulatory compound, such as lenalidomide, is administered no more than about 96 hours, 72 hours, 48 hours, 24 hours, 12 hours, 6 hours, 2 hours, or 1 hour before the start of T cell therapy.

[0338] In some such embodiments, in which an immunomodulatory compound, e.g., lenalidomide, is administered prior to cell therapy (e.g., T cell therapy, such as CAR T cell therapy), administration of the immunomodulatory compound, e.g., lenalidomide, is continued at regular intervals before the start of cell therapy and/or for some time after the start of cell therapy.

[0339] In some embodiments, an immunomodulatory compound, eg, lenalidomide, is administered or continues to be administered following the administration of cell therapy (eg, T cell therapy, such as CAR T cell therapy). In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered within or about within 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 hours, 96 hours, 4 days, 5 days, 6 days, or 7 days, 14 days, 15 days, 21 days, 24 days, 28 days, 30 days, 36 days, 42 days, 60 days, 72 days, or 90 days after starting cell therapy (eg, T-cell therapy,). In some embodiments, the proposed methods include continuously administering, for example, at regular intervals, an immunomodulatory compound after the start of a cell therapy administration.

[0340] In some embodiments, the immunomodulatory compound, e.g., lenalidomide, is administered before or about 1 day, before or about 2 days, before or about 3 days, before or about 4 days, before or about 5 days up to or up to about 6 days, up to or up to about 7 days, up to or up to about 12 days, up to or up to about 14 days, up to or up to about 21 days, up to or up to about 24 days, up to or up to about 28 days, up to or up to about 30 days, up to or up to about 35 days, up to or up to about 42 days, up to or up to about 60 days, or up to or up to about 90 days, up to or up to about 120 days, up to or up to about 180 days, up to or up to about 240 days, up to or up to about 360 days, or up to or up to about 720 days or more after the start of the administration of cell therapy (eg, T cell therapy, such as CAR T cell therapy).

[0341] In some of the above embodiments, an immunomodulatory compound, eg, lenalidomide, is administered before and after the start of the cell therapy (eg, T cell therapy, such as CAR T cell therapy).

[0342] In some embodiments, the start of administration of an immunomodulatory compound, for example, lenalidomide, is carried out at or after a time point, optionally immediately after or within 1-3 days after: (i) a peak maximum is detected in the subject's blood cell levels from T-cell therapy; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

[0343] In some embodiments, the initiation of administration of an immunomodulatory compound, for example, lenalidomide, in at least one cycle occurs after the initiation of administration of T-cell therapy. In some embodiments, the onset of administration of an immunomodulatory compound, such as lenalidomide, occurs when at least, or about, at least 1 day, at least, or about, at least 2 days, at least, or about, at least 3 days, at least, or about, at least 4 days, at least, or about at least 5 days, at least, or about at least 6 days, at least, or about at least 7 days, at least, or about, at least 8 days, at least, or about, at least 9 days, at least, or about, at least 10 days, at least, or about, at least at least 12 days at least or about at least 14 days at least or about at least 15 days at least or about at least 21 days at least , or at least about 24 days, at least, or about at least 28 days, at least, or about at least 30 days, at least, or about at least at least 35 days or at least or about at least 42 days, at least or about at least 60 days, or at least or about at least 90 days days after initiation of T-cell therapy. In some embodiments, the start of administration of an immunomodulatory compound, such as lenalidomide, is at least 2 days later, at least 1 week later, at least 2 weeks later, at least 3 weeks later, or at least at least 4 weeks after the start of the introduction of T-cell therapy. In some embodiments, the start of administration of an immunomodulatory compound, such as lenalidomide, is 2-28 or 7-21 days after the start of T-cell therapy. In some embodiments, the start of the administration of an immunomodulatory compound, for example, lenalidomide, is carried out at a time point that occurs more than or about more than 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 24 days, or 28 days after initiation of T-cell therapy. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered several times a day, twice a day, daily, every other day, three times a week, twice a week, or once a week after starting cell therapy. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered daily. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered twice a day. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered three times a day. In other embodiments, an immunomodulatory compound, such as lenalidomide, is administered every other day. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered daily. In some embodiments, an immunomodulatory compound, e.g., lenalidomide, is administered over a period of administration over multiple consecutive days, e.g., up to about 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more than 30 consecutive days. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered for more than or about more than 7 consecutive days, more than or about more than 14 consecutive days, more than or about more than 21 consecutive days, or more than or about more than 28 consecutive days. In some embodiments, an immunomodulatory compound, eg, lenalidomide, is administered during an administration period of up to 21 days, sequentially. In some embodiments, the immunomodulatory compound, for example, lenalidomide, is administered during an administration period of up to 21 days, sequentially, where the cycle includes more than 30 days from the start of administration of the immunomodulatory compound.

[0344] In some embodiments, an immunomodulatory compound, e.g., lenalidomide, is administered during an administration period of no more than about 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or no more than 30 consecutive days. In certain embodiments, lenalidomide is administered once a day for 14 days during a 21 day treatment cycle. In certain embodiments, lenalidomide is administered once daily for 21 days during a 28 day treatment cycle. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered during an administration period of no more than 14 consecutive days.

[0345] In some embodiments, the immunomodulatory compound, e.g., lenalidomide, is administered in a cycle, where the cycle comprises administering the immunomodulatory compound, e.g., lenalidomide, for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered. In some embodiments, the rest period is greater than about 1 day, greater than about 3 consecutive days, greater than about 5 consecutive days, greater than about 7 consecutive days, greater than about 8 consecutive days, greater than about 9 consecutive days, more than about 10 consecutive days, more than about 11 consecutive days, more than about 12 consecutive days, more than about 13 consecutive days, more than about 14 consecutive days, more than about 15 consecutive days more than about 16 consecutive days, more than about 17 consecutive days, more than about 18 consecutive days, more than about 19 consecutive days, more than about 20 consecutive days, or more than about 21 or more consecutive days. In some embodiments, the rest period is greater than 7 consecutive days, greater than 14 consecutive days, greater than 21 days, or greater than 28 days. In some embodiments, the rest period is greater than about 14 consecutive days. In some embodiments, the immunomodulatory compound administration cycle does not include a rest period.

[0346] In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered in a cycle where the cycle is repeated at least once. In some embodiments, an immunomodulatory compound, e.g., lenalidomide, is administered for at least 2 cycles, at least 3 cycles, at least 4 cycles, at least 5 cycles, at least 6 cycles , at least 7 cycles, at least 8 cycles, at least 9 cycles, at least 10 cycles, at least 11 cycles, or at least 12 cycles. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23 or 24 cycles.

[0347] In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered six times a day, five times a day, four times a day, three times a day, twice a day, once a day, every other day, every three days, twice a week, once a week, or only once before or after the start of T-cell therapy. In some embodiments, the immunomodulatory compound, eg, lenalidomide, is administered at multiple doses at regular intervals before, during, during, and/or after the T cell therapy administration period. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered at one or more doses at regular intervals prior to the administration of T cell therapy. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered at one or more doses at regular intervals following the administration of T cell therapy. In some embodiments, one or more dosages of an immunomodulatory compound, such as lenalidomide, may be administered concurrently with a dose of T cell therapy.

[0348] In some embodiments, the dose, frequency, duration, timing, and/or order of administration of an immunomodulatory compound, such as lenalidomide, is determined based on specific thresholds or criteria, the results of the screening step and/or evaluation of treatment outcomes described in herein, such as those described in Section III of this document.

[0349] In some embodiments, the method includes administering the cell therapy to a subject who has previously received a therapeutically effective amount of an immunomodulatory compound. In some embodiments, the immunomodulatory compound is administered to the subject prior to administration of a dose of cells expressing the recombinant receptor in the subject. In some embodiments, the treatment with the immunomodulatory compound occurs at the same time as the administration of the cell dose. In some embodiments, the immunomodulatory compound is administered following a dose of cells.

[0350] In some embodiments, an immunomodulatory compound, e.g., lenalidomide, is administered daily for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 days, or more. In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered twice a day for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 days or more . In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered three times a day for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 days or more . In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered every other day for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 days or more.

[0351] In some embodiments of the methods provided herein, an immunomodulatory compound, such as lenalidomide, and T cell therapy are administered simultaneously or nearly simultaneously.

[0352] In some embodiments, the immunomodulatory compound, e.g., lenalidomide, is administered at a dose of from or about 0.1 mg to about 100 mg, from or about 0.1 mg to 50 mg, from or about 0.1 mg to 25 mg, from or about 0.1 mg to 10 mg, from or about 0.1 mg to 5 mg, from or about 0.1 mg to 1 mg, from or about 1 mg to 100 mg , from or about 1 mg to 50 mg, from or about 1 mg to 25 mg, from or about 1 mg to 10 mg, from or about 1 mg to 5 mg, from or about 5 mg to 100 mg , from or about 5 mg to 50 mg, from or about 5 mg to 25 mg, from or about 5 mg to 10 mg, from or about 10 mg to 100 mg, from or about 10 mg to 50 mg , from or from 10 mg to 25 mg, from or from about 25 mg to 100 mg, from or from about 25 mg to 50 mg, or from or from about 50 mg to 100 mg, each value inclusive. In some embodiments, this amount is a daily administered amount of an immunomodulatory compound, such as lenalidomide.

[0353] In some embodiments, the immunomodulatory compound, e.g., lenalidomide, is administered at a dose of about 1 mg to about 20 mg, e.g., about 1 mg to about 10 mg, about 2.5 mg to about 7.5 mg, about 5 mg to about 15 mg, e.g., about 5 mg, 10 mg , 15 mg or 20 mg. In some embodiments, lenalidomide is administered at a dose of from about 10 µg/kg to 5 mg/kg, for example, about 100 µg/kg to about 2 mg/kg, about 200 µg/kg to about 1 mg/kg, about 400 µg /kg to about 600 µg/kg, for example, about 500 µg/kg. In some embodiments, this amount is a daily administered amount of an immunomodulatory compound, such as lenalidomide.

[0354] In some embodiments, an immunomodulatory compound, e.g., lenalidomide, is administered at a total daily dosage amount of at least, or about, at least 0.1 mg per day, 0.5 mg per day, 1.0 mg per day, 2.5 mg per day, 5 mg per day, 10 mg per day, 25 mg per day, 50 mg per day, or 100 mg per day. In some embodiments, the dose of lenalidomide is or about 25 mg per day. In specific embodiments, the dose of lenalidomide is or about 10 mg per day.

[0355] In some embodiments, the immunomodulatory compound, e.g., lenalidomide, is administered in an amount greater than or about greater than 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg, and less than 25 mg. In some embodiments, the immunomodulatory compound, e.g., lenalidomide, is administered in an amount greater than or about greater than 1 mg per day, 2.5 mg per day, 5 mg per day, 7.5 mg per day, 10 mg per day, 15 mg per day and less than 25 mg per day.

[0356] In any of the above embodiments, an immunomodulatory compound, such as lenalidomide, may be administered orally. In some embodiments, the immunomodulatory compound, such as lenalidomide, is administered as a tablet or capsule.

[0357] In some embodiments, dosages, such as a daily dosage, are administered in one or more divided doses, such as 2, 3, or 4 doses, or in a single preparation. An immunomodulatory compound, such as lenalidomide, may be administered alone in the presence of a pharmaceutically acceptable carrier or in the presence of other therapeutic agents.

[0358] It is understood that higher or lower doses of the immunomodulatory compound may be used, for example, depending on the particular agent and route of administration. In some embodiments, the immunomodulatory compound may be administered alone or in the form of a pharmaceutical composition, wherein the compound is present in an additive or admixture with one or more pharmaceutically acceptable carriers, excipients, or diluents. In some embodiments, the immunomodulatory compound may be administered either systemically or locally to the organ or tissue to be treated. Exemplary routes of administration include, but are not limited to, external, injection (such as subcutaneous, intramuscular, intradermal, intraperitoneal, intratumoral, and intravenous), oral, sublingual, rectal, transdermal, intranasal, vaginal, and inhalation routes. In some embodiments, the route of administration is oral, parenteral, rectal, nasal, topical, or ocular, or inhalation. In some embodiments, the immunomodulatory compound is administered orally. In some embodiments, the immunomodulatory compound is administered orally in solid dosage forms such as capsules, tablets and powders, or in liquid dosage forms such as elixirs, syrups and suspensions.

[0359] After improvement in the patient's condition, the dose may be adjusted for preventive or maintenance treatment. For example, the dose or frequency of administration, or both, may be reduced depending on the symptoms, to a level at which the desired therapeutic or prophylactic effect is maintained. If symptoms improve to an appropriate level, treatment may be discontinued. However, patients may require intermittent treatment on a long-term basis for any recurrence of symptoms. Patients may also require chronic treatment on a long-term basis.

C. Antilymphocyte treatment

[0360] In some aspects, the proposed methods may further include the introduction of one or more types of antilymphocyte therapy, for example, before or simultaneously with the start of the introduction of T-cell therapy. In some embodiments, the anti-lymphocyte therapy comprises the administration of a phosphamide, such as cyclophosphamide. In some embodiments, the anti-lymphocyte therapy may include the administration of fludarabine.

[0361] In some aspects, pre-treatment of subjects with immunodepletion (eg, anti-lymphocyte) therapy may improve the effects of adoptive cell therapy (ACT). Pretreatment with anti-lymphocyte agents, including combinations of cyclosporine and fludarabine, is effective in improving the efficacy of tolerated antitumor effector lymphocytes (TILs) in cell therapy, including improving the response and/or persistence of transferred cells. See, for example, Dudley et al., Science, 298, 850-54 (2002); Rosenberg et al., Clin Cancer Res, 17(13):4550-4557 (2011). Similarly, in the context of CAR+ T lymphocytes, several studies have included anti-lymphocyte agents, most commonly cyclophosphamide, fludarabine, bendamustine, or combinations thereof, sometimes accompanied by low doses of radiation. See Han et al. Journal of Hematology & Oncology, 6:47 (2013); Kochenderfer et al., Blood, 119: 2709-2720 (2012); Kalos et al., Sci Transl Med, 3(95):95ra73 (2011); Clinical Trial Study Record No.: NCT02315612; NCT01822652.

[0362] Such pre-training may be carried out in order to reduce the risk of one or more of the various results that could reduce the effectiveness of therapy. These include a phenomenon known as "cytokine sink" in which T-lymphocytes, B-lymphocytes, NK cells compete with TIL for homeostatic and activating cytokines such as IL-2, IL-7 and/or IL-15; suppression of TIL by regulatory T lymphocytes, NK cells, or other cells of the immune system; impact of negative regulators on the tumor microenvironment. Muranski et al., Nat Clin Pract Oncol. December; 3(12): 668-681 (2006).

[0363] Thus, in some embodiments, the proposed method further comprises administering anti-lymphocyte therapy to the subject. In some embodiments, the method includes administering an anti-lymphocyte therapy to a subject prior to administration of a dose of cells. In some embodiments, the anti-lymphocyte therapy comprises a chemotherapeutic agent such as fludarabine and/or cyclophosphamide. In some embodiments, administration of cells and/or anti-lymphocyte therapy is via in vitro delivery.

[0364] In some embodiments, the methods include administering a pretreatment agent, such as an anti-lymphocyte or chemotherapeutic agent, such as cyclophosphamide, fludarabine, or combinations thereof, to the subject prior to administration of the cell dose. For example, the pretreatment agent may be administered to the subject at least 2 days, such as at least 3, 4, 5, 6, or 7 days prior to the first or subsequent dosing. In some embodiments, the pretreatment agent is administered to the subject no more than 7 days, such as no more than 6, 5, 4, 3, or 2 days prior to cell dosing.

[0365] In some embodiments, the subject is pretreated with cyclophosphamide at a dose between or about 20 mg/kg and 100 mg/kg, such as between or about 40 mg/kg and 80 mg /kg. In some aspects, the subject is pretreated with (or about) 60 mg/kg cyclophosphamide. In some embodiments, fludarabine may be administered in a single dose or may be administered in multiple doses, such as daily, every other day, or every three days. In some embodiments, the implementation, cyclophosphamide is administered once a day for one or two days.

[0366] In some embodiments, when the antilymphocyte agent comprises fludarabine, fludarabine is administered to the subject at a dose between or about 1 mg/m ² and 100 mg/m ² , such as between or about 10 mg / m ² and 75 mg / m ² , 15 mg / m ² and 50 mg / m ² , 20 mg / m ² and 30 mg / m ² , or 24 mg / m ² and 26 mg / m ² . In some cases, the subject is administered 25 mg/m ² fludarabine. In some embodiments, fludarabine may be administered in a single dose or may be administered in multiple doses, such as daily, every other day, or every three days. In some embodiments, fludarabine is administered daily, eg for 1-5 days, eg for 3-5 days.

[0367] In some embodiments, the anti-lymphocyte agent comprises a combination of agents, such as a combination of cyclophosphamide and fludarabine. Thus, the combination of agents may contain cyclophosphamide at any dosing or administration schedule, eg, as described above, and fludarabine at any dosage or administration schedule, eg, as described above. For example, in some aspects, prior to cell dosing, the subject is administered 60 mg/kg (~2 g/m ² ) cyclophosphamide and 3-5 doses of 25 mg/m ² fludarabine.

[0368] In one exemplary dosing regimen, before receiving the first dose, the subject receives an immunomodulatory compound 1 day prior to cell administration and antilymphocyte chemotherapy of cyclophosphamide and fludarabine (CY/FLU) for pretreatment, which is administered at least two days prior to the first dose of CAR-expressing cells and, as a rule, no more than 7 days before the introduction of cells. In another exemplary dosing regimen, subjects receive the immunomodulatory compound at the same time as the cells are administered, eg, on the same day. In yet another exemplary dosing regimen, subjects receive the immunomodulatory compound several days after cell administration, eg, 7, 8, 9, 10, 11, 12, 13, 14, or more than 14 days. In some cases, for example, cyclophosphamide is given 24-27 days after the administration of an immunomodulatory compound, for example, lenalidomide. After pretreatment, subjects are dosed with CAR-expressing T-lymphocytes as described above.

[0369] In some embodiments, administration of a pretreatment agent prior to cell dose infusion improves treatment outcome. For example, in some aspects, the pretreatment improves the efficacy of treatment with a dose or increases the persistence of cells expressing the recombinant receptor (eg, CAR-expressing cells, such as CAR-expressing T lymphocytes) in a subject. In some embodiments, pretreatment increases disease survival, such as the percentage of subjects who survive and do not exhibit minimal residual or molecularly detectable disease after a given period of time after cell dosing. In some embodiments, the median survival time after illness is increased.

[0370] After the cells are introduced into a subject (eg, a human), the biological activity of populations of genetically engineered cells, in some aspects, is measured using any of the numerous known methods. Evaluation parameters include specific binding of engineered or natural T-lymphocytes or other immune cells to an antigen, in vivo, eg, by imaging, or ex vivo, eg, by ELISA or flow cytometry. In certain embodiments, the ability of engineered cells to destroy target cells can be measured using any appropriate method known in the art, such as cytotoxicity assays, as described, for example, in Kochenderfer et al., J. Immunotherapy, 32(7): 689 -702 (2009) and Herman et al. J. Immunological Methods, 285(1): 25-40 (2004). In certain embodiments, the biological activity of cells can also be measured by assaying the expression and/or secretion of certain cytokines such as CD107a, IFNγ, IL-2, and TNF. In some aspects, biological activity is measured by assessing clinical outcomes of treatment, such as reduction in tumor mass or burden. In some aspects, the toxic effects of the treatment, the persistence and/or proliferation of cells, and/or the presence or absence of a host immune response are assessed.

[0371] In some embodiments, administration of a pretreatment agent prior to cell dose infusion improves treatment outcome, e.g., by improving treatment efficacy with that dose, or increases the persistence of cells expressing the recombinant receptor (e.g., CAR-expressing cells, such as CAR-expressing T-lymphocytes) in a subject. Therefore, in some embodiments, the dose of the pretreatment agent given in a method that is combination therapy with an immunomodulatory compound and cell therapy is higher than the dose given in a method without an immunomodulatory compound.

II. T-cell therapy and genetically engineered cells

[0372] In some embodiments, T-cell therapy for use in conjunction with the proposed methods of combination therapy includes the introduction of genetically engineered cells expressing recombinant receptors designed to recognize and/or specifically bind molecules associated with a disease or condition, and they give resulting in a reaction, such as an immune reaction, against such molecules upon binding to such molecules. Receptors may include chimeric receptors, such as chimeric antigen receptors (CARs), and other transgenic antigen receptors, including transgenic T-lymphocyte receptors (TCRs).

[0373] In some embodiments, cells comprise or are derived from engineered cells such that they contain a genetically engineered receptor, such as a genetically engineered antigen receptor, such as a chimeric antigen receptor (CAR) or a T-lymphocyte receptor (TCR). Also provided are populations of such cells, compositions containing such cells and/or enriched with such cells, for example, in which cells of a certain type, such as T-lymphocytes or CD8+ or CD4+ cells, are enriched or selected. Among the compositions are pharmaceutical compositions and preparations for administration, for example, for adoptive cell therapy. Also provided are therapeutic methods for administering cells and compositions to subjects, such as patients.

[0374] Thus, in some embodiments, cells contain one or more genetically engineered nucleic acids and express recombinant or genetically engineered products of such nucleic acids. In some embodiments, gene transfer is accomplished by first stimulating cells, e.g., by combining them with a stimulus that induces a response, such as proliferation, survival, and/or activation, e.g., as measured by expression of a cytokine or an activation marker, followed by transduction activated cells and propagation in culture to quantities sufficient for clinical applications.

A. Recombinant receptors

[0375] Cells typically express recombinant receptors, such as antigen receptors, including functional non-TCR antigen receptors, such as chimeric antigen receptors (CAR), and other antigen-binding receptors, such as transgenic T-lymphocyte receptors (TCR ). Also among the receptors are other chimeric receptors.

1. Chimeric antigen receptors (CARs)

[0376] In some embodiments, engineered cells, such as T-lymphocytes used in the proposed embodiments, express CAR with specificity for a particular antigen (or marker or ligand), such as an antigen expressed on the surface of a particular cell type. In some embodiments, the antigen is a polypeptide. In some embodiments, it is a carbohydrate or other molecule. In some embodiments, the antigen is selectively expressed or overexpressed on cells of the disease or condition, such as tumor cells or pathogenic cells, over normal or non-target cells or tissues. In other embodiments, the antigen is expressed on normal cells and/or is expressed on genetically engineered cells.

[0377] In specific embodiments, the recombinant receptor, such as a chimeric receptor, contains an intracellular signaling region that contains a cytoplasmic signaling domain or a region (also interchangeably referred to as an intracellular signaling domain or region), such as a cytoplasmic (intracellular) region capable of inducing a primary activation in a T-lymphocyte, e.g., a cytoplasmic signaling domain or a T-lymphocyte receptor (TCR) component region (e.g., a cytoplasmic signaling domain or a CD3-zeta (CD3ζ) zeta chain region or a functional variant or a signaling portion thereof) and/or which contains an immunoreceptor tyrosine activating motif (ITAM).

[0378] In some embodiments, the chimeric receptor further comprises an extracellular ligand-binding domain that specifically binds to an antigen (eg, antigen) ligand. In some embodiments, the chimeric receptor is a CAR that contains an extracellular antigen recognition domain that specifically binds to an antigen. In some embodiments, the ligand, such as an antigen, is a protein expressed on the surface of cells.

[0379] In some embodiments, the CAR is a TCR-like CAR and the antigen is a truncated peptide antigen, such as an intracellular protein peptide antigen that, like TCR, is recognized on the cell surface in the context of a major histocompatibility complex (MHC) molecule. Generally, a CAR containing an antibody or antigen-binding fragment that exhibits TCR-like specificity against peptide-MHC complexes may also be referred to as a TCR-like CAR. In some embodiments, the extracellular antigen-binding domain specific for the MHC-peptide TCR-like CAR complex is associated with one or more intracellular signaling components, in some aspects via linkers and/or transmembrane domain(s). In some embodiments, such molecules generally can signal, or approximately reproduce, a signal through a natural antigen receptor, such as a TCR, and optionally, a signal through such a receptor in combination with a co-stimulatory receptor.

[0380] Exemplary antigen receptors, including CARs, and methods for genetically engineering and introducing such receptors into cells include those described, for example, in International Patent Applications Nos. /071154, WO2013/123061, WO2016/0046724, WO2016/014789, WO2016/090320, WO2016/094304, WO2017/025038, WO2017/173256, публикациях заявок на патент США номера US2002131960, US2013287748, US20130149337, в патентах США №№ 6451995, 7446190, 8252592, 8339645, 8398282, 7446179, 6410319, 7070995, 7265209, 7354762, 7446191, 8324353, 8479118 и 9765342 и в заявке на Европейский патент номер EP2537416, и/или способы, описанные Sadelain et al., Cancer Discov., 3 (4): 388-398 (2013); Davila et al., PLoS ONE 8(4): e61338 (2013); Turtle et al., Curr. Opin. Immunol., 24(5): 633-39 (2012); Wu et al., Cancer, 18(2): 160-75 (2012). In some aspects, antigen receptors include CARs as described in US Pat. No. 7,446,190 and those described in International Patent Application Publication No. WO/2014055668 A1. Examples of CARs include CARs as described in any of the publications discussed above such as WO2014031687, US Patent No. 8339645, US Patent No. 7446179, US Patent No. 2013/0149337, US Patent No. 7446190, US Patent No. 8389282, Kochenderfer et al. , Nature Reviews Clinical Oncology, 10, 267-276 (2013); Wang et al., J. Immunother. 35(9): 689-701 (2012); and Brentjens et al., Sci Transl Med. 5(177) (2013). See also WO2014031687, US Patent No. 8339645, US Patent No. 7446179, US Patent No. 2013/0149337, US Patent No. 7446190 and US Patent No. 8389282. Chimeric receptors, such as CAR, typically contain an extracellular antigen-binding domain, such as part of an antibody molecule, typically the heavy chain variable region (V _H ) and/or the light chain variable region (V _L ) of an antibody, eg, a fragment of an scFv antibody.

[0381] In some embodiments, CARs are designed with specificity for a particular antigen (or marker or ligand), such as an antigen expressed in a particular cell type to which adoptive therapy is to be targeted, for example, a cancer marker and/or an antigen targeted to induce a damping response, such as an antigen expressed on a normal or non-diseased cell type. Thus, a CAR typically contains in its extracellular portion one or more antigen-binding molecules, such as one or more antigen-binding fragments, domains, or moieties, or one or more antibody variable domains and/or antibody molecules. In some embodiments, the CAR comprises an antigen-binding antibody moiety or moieties, such as a single chain antibody fragment (scFv) derived from the variable heavy (V _H ) and variable light (V _L ) chains of a monoclonal antibody (mAb), or a single domain antibody. (sdAb) such as sdFv, nanobody, V _H H and V _NAR. In some embodiments, the antigen-binding fragment contains the variable regions of the antibody connected by a flexible linker.

[0382] Among the antigen-binding domains included in CARs are antibody fragments. “Antibody fragment” or “antigen-binding fragment” refers to a molecule, other than an intact antibody, that contains a portion of the intact antibody that binds the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab') ₂ ; diabody; linear antibodies; heavy chain variable regions (V _H ), single chain antibody molecules such as scFv, and single domain antibodies containing only the V _H region; and multispecific antibodies formed from antibody fragments. In specific embodiments, the antibodies are single chain antibody fragments containing a heavy chain variable region (V _H ) and/or a light chain variable region (V _L ), such as scFv.

[0383] In certain embodiments, multispecific binding molecules, e.g., multispecific chimeric receptors, such as multispecific CARs, may comprise any of the multispecific antibodies, including, e.g., bispecific antibodies, multispecific single chain antibodies, e.g., diabodies, tribodies, and tetrabodies, tandem di-scFv and tri-scFv tandem.

[0384] Single domain antibodies (sdAbs) are antibody fragments containing all or part of the antibody heavy chain variable region or all or part of the light chain variable region. In certain embodiments, the single domain antibody is a human single domain antibody.

[0385] Antibody fragments can be obtained using various techniques, including, but not limited to, proteolytic fermentation of intact antibodies, as well as production by recombinant host cells. In some embodiments, antibodies are recombinantly produced fragments, such as fragments containing moieties that do not occur naturally, such as fragments with two or more antibody regions or chains connected by synthetic linkers, e.g., peptide linkers, and/or those that cannot be obtained by enzymatic digestion of a naturally occurring intact antibody. In some aspects, the antibody fragments are scFv.

[0386] In some embodiments, the antibody or antigen-binding fragment thereof is a single chain antibody fragment, such as a single chain variable fragment (scFv) or a diabody or single domain antibody (sdAb). In some embodiments, the antibody or antigen binding fragment is a single domain antibody containing only the V _H region. In some embodiments, the antibody or antigen-binding fragment is an scFv containing a heavy chain variable region (V _H ) and a light chain variable region (V _L ).

[0387] In some embodiments, the antigen targeted by the receptor is a polypeptide. In some embodiments, it is a carbohydrate or other molecule. In some embodiments, the antigen is selectively expressed or overexpressed on cells of the disease or condition, such as tumor or pathogenic cells, over normal or non-target cells or tissues. In other embodiments, the antigen is expressed on normal cells and/or is expressed on genetically engineered cells.

[0010] In certain embodiments, the antigen is or contains an αvβ6 integrin (avb6 integrin), B-lymphocyte maturation antigen (BCMA), B7-H3, B7-H6, carbonic anhydrase 9 (CA9, also known as CAIX or G250) , testicular cancer antigen, testicular cancer antigen 1B (CTAG, also known as NY-ESO-1 and LAGE-2), carcinoembryonic antigen (CEA), cyclin, cyclin A2, chemokine motif C-C ligand 1 (CCL-1), CD19, CD20, CD22, CD23, CD24, CD30, CD33, CD38, CD44, CD44v6, CD44v7/8, CD123, CD133, CD138, CD171, chondroitin sulfate proteoglycan 4 (CSPG4), epidermal growth factor protein (EGFR), truncated epidermal growth factor protein (tEGFR), epidermal growth factor receptor type III (EGFR vIII) mutation, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), ephrin B2, ephrin A2 receptor (EPHa2), estrogen receptor, Fc receptor like 5 (FCRL5; also known as Fc receptor homologue 5 or FCRH5), fetal acetylcholine receptor (fetal AchR), folate binding protein (FBP), folate receptor alpha, ganglioside GD2, O-acetylated GD2 (OGD2), ganglioside GD3, glycoprotein 100 (gp100), glypican-3 (GPC3), G protein-coupled 5D receptor (GPCR5D), Her2/neu (erb-B2 tyrosine kinase receptor), Her3 (erb-B3), Her4 (erb-B4), erbB dimers , high molecular weight human melanoma-associated antigen (HMW-MAA), hepatitis B surface antigen, human leukocyte A1 antigen (HLA-A1), human leukocyte A2 antigen (HLA-A2), IL-22 receptor alpha (IL-22Rα), IL-13 alpha 2 receptor (IL-13Rα2), kinase insert domain containing receptor (kdr), kappa light chain, cell adhesion molecule L1 (L1-CAM), CE7 epitope L1-CAM,leucine-enriched insert containing element A of family 8 (LRRC8A), Ley antigen, melanoma associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, MAGE-A10, mesothelin (MSLN), c-Met, mouse cytomegalovirus (CMV), mucin 1 (MUC1) , MUC16, natural killer group 2 D element ligands (NKG2D), melan A (MART-1), neural cell adhesion molecule (NCAM), oncofetal antigen, preferably expressed melanoma antigen (PRAME), progesterone receptor, prostate-specific antigen, antigen prostate stem cell (PSCA), prostate-specific membrane antigen (PSMA), tyrosine kinase receptor like orphan receptor 1 (ROR1), survivin, trophoblast glycoprotein (TPBG also known as 5T4), tumor-associated glycoprotein 72 (TAG72), protein 1 tyrosinase related protein (TRP1, also known as TYRP1 or gp75), tyrosinase related protein 2 (TRP2, also known as dopachrom tautomerase, dopachrome delta isomerase or DCT), vascular endothelial growth factor receptor (VEGFR), vascular endothelial growth factor receptor 2 (VEGFR 2), Wilms tumor 1 (WT-1), pathogen-specific or pathogen-expressed antigen or universal label-associated antigen, and/or biotinylated molecules and/or molecules expressed by HIV, HCV, HBV or other pathogens. The antigens targeted by the receptors, in some embodiments, include antigens associated with B-lymphocyte cancer, such as any of a variety of known B-lymphocyte markers. In some embodiments, the antigen is or comprises CD20, CD19, CD22, ROR1, CD45, CD21, CD5, CD33, Igkappa, Iglambda, CD79a, CD79b, or CD30.

[0388] In some embodiments, the antigen is or contains a pathogen-specific or pathogen-expressed antigen. In some embodiments, the antigen is a viral antigen (such as a viral antigen from HIV, HCV, HBV, and the like), bacterial antigens, and/or parasitic antigens.

[0389] In some embodiments, the CAR is an anti-BCMA CAR that is specific for BCMA, eg, human BCMA. Chimeric antigen receptors containing anti-BCMA antibodies, including mouse anti-human BCMA antibodies and human anti-human BCMA antibodies and cells expressing such chimeric receptors have been previously described. Watch Carpenter et al., Clin Cancer Res., 2013, 19 (8): 2048-2060, US 9765342, Wo 2016/090320, Wo2016090327, Wo2010104949A2, Wo2016/0046724, Wo2016/014789, Wo2016/09430, Wo2017/0258 and WO2017173256. In some embodiments, the anti-BCMA CAR contains an antigen-binding domain, such as an scFv containing a variable heavy (V_H) and/or variable light (V_L) a region derived from an antibody described in WO 2016/090320 or WO2016090327. In some embodiments, the antigen binding domain is an antibody fragment containing a heavy chain variable region (V_H) and the light chain variable region (V_L). In some aspects, the area, V_H is or contains an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with the region V amino acid sequence_H, listed in any of SEQ ID NOs: 30, 32, 34, 36, 38, 40, 42, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103 , 105, 107, 109, 111, 181, 183, 185 and 188; and/or region V_L is or includes an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the region V amino acid sequence_L, listed in any of SEQ ID NOs: 31, 33, 35, 37, 39, 41, 43, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104 , 106, 108, 110, 112, 182, 184, 186 and 189.

[0390] In some embodiments, the antigen-binding domain, such as scFv, contains V _H as shown in SEQ ID NO: 30 and V _L as shown in SEQ ID NO: 31. In some embodiments, the antigen-binding domain, such as scFv, contains the V _H shown in SEQ ID NO: 32 and the V _L shown in SEQ ID NO: 33. In some embodiments, an antigen-binding domain, such as scFv, comprises V _H as shown in SEQ ID NO: 34 and V _L as shown in SEQ ID NO: 35. In some embodiments, an antigen-binding domain, such as scFv contains V _H as shown in SEQ ID NO: 36 and V _L as shown in SEQ ID NO: 37. In some embodiments, an antigen-binding domain, such as scFv, comprises V _H as shown in SEQ ID NO: 38 and V _L as shown in SEQ ID NO: 39. In some embodiments, an antigen-binding domain, such as scFv, contains V _H as shown in SEQ ID NO: 40 and V _L as shown in SEQ ID NO: 41. In some embodiments, an antigen binding domain, such as scFv, contains V _H as shown in SEQ ID NO: 42, and V _L as shown in SEQ ID NO: 43. In some embodiments, the antigen binding domain, such as scFv, comprises V _H as shown in SEQ ID NO: 77 and V _L as shown in SEQ ID NO: 78. B in some embodiments, an antigen binding domain, such as scFv, comprises V _H as shown in SEQ ID NO: 79 and V _L as shown in SEQ ID NO: 80. In some embodiments, an antigen binding domain, such as scFv , contains V _H as shown in SEQ ID NO: 81 and V _L as shown in SEQ ID NO: 82. In some embodiments, a An antigen binding domain, such as scFv, contains V _H as shown in SEQ ID NO: 83 and V _L as shown in SEQ ID NO: 84. In some embodiments, an antigen binding domain, such as scFv, contains V _H , shown in SEQ ID NO: 85, and V _L shown in SEQ ID NO: 86. In some embodiments, the antigen-binding domain, such as scFv, contains V _H shown in SEQ ID NO: 87, and V _L shown in SEQ ID NO: 88. In some embodiments, the antigen-binding domain, such as scFv, contains V _H shown in SEQ ID NO: 89, and V _L shown in SEQ ID NO: 90. In some embodiments, implementation, the antigen-binding domain, such as scFv, contains V _H shown in SEQ ID NO: 91, and V _L shown in SEQ ID NO: 92. In some embodiments, the antigen-binding domain, such as scFv, contains V _H as shown in SEQ ID NO: 93 and V _L as shown in SEQ ID NO: 94. In some embodiments, an antigen binding domain such as to scFv contains V _H as shown in SEQ ID NO: 95 and V _L as shown in SEQ ID NO: 96. In some embodiments, an antigen-binding domain, such as scFv, contains V _H as shown in SEQ ID NO: : 97, and V _L as shown in SEQ ID NO: 98. In some embodiments, the antigen-binding domain, such as scFv, contains V _H as shown in SEQ ID NO: 99 and V _L as shown in SEQ ID NO: : 100. In some embodiments, the antigen-binding domain, such as scFv, comprises V _H as shown in SEQ ID NO: 101 and V _L as shown in SEQ ID NO: 102. In some embodiments, the antigen-binding domain , such as scFv, contains the V _H shown in SEQ ID NO: 103 and the V _L shown in SEQ ID NO: 104. In some embodiments, an antigen-binding domain, such as scFv, contains the V _H shown in SEQ ID NO: 105, and V _L shown in SEQ ID NO: 106. In some embodiments, the antigen binding domain, such as scFv, contains V _H , as shown in SEQ ID NO: 107 and V _L as shown in SEQ ID NO: 106. In some embodiments, the antigen binding domain, such as scFv, contains the V _H as shown in SEQ ID NO: 30 and V _L , shown in SEQ ID NO: 108. In some embodiments, an antigen binding domain, such as scFv, contains V _H shown in SEQ ID NO: 109 and V _L shown in SEQ ID NO: 110. In some embodiments, implementation , an antigen binding domain such as scFv contains V _H as shown in SEQ ID NO: 111 and V _L as shown in SEQ ID NO: 112. In some embodiments, an antigen binding domain such as scFv contains V _H as shown in SEQ ID NO: 181 and V _L as shown in SEQ ID NO: 182. In some embodiments, an antigen binding domain such as scFv contains V _H as shown in SEQ ID NO: 183 and V _L as shown in SEQ ID NO: 184. In some embodiments, the antigen binding domain, such as scFv, contains V _H as shown in SEQ ID NO: 18 5 and V _L as shown in SEQ ID NO: 186. In some embodiments, the antigen binding domain, such as scFv, comprises V _H as shown in SEQ ID NO: 187 and V _L as shown in SEQ ID NO: 188. In some embodiments, V _H or V _L has an amino acid sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to any of the above V _H or V _L sequences and retains binding to BCMA. In some embodiments, the V _H region is at the amino terminus of the V _L region. In some embodiments, the V _H region is at the carboxy terminus of the V _L region. In some embodiments, the variable heavy and variable light chains are connected by a linker. In some embodiments, the linker is listed in SEQ ID NO: 70, 72, 73, 74, or 189.

[0391] In some embodiments, the CAR is an anti-CD19 CAR that is specific for CD19, eg, human CD19. In some embodiments, the implementation of the antigen-binding domain contains V _H and/or V _L derived from FMC63, which, in some aspects, may be scFv. In some embodiments, the scFv and/or V _H domains are derived from FMC63. FMC63 generally refers to a mouse IgG1 monoclonal antibody raised against Nalm-1 and -16 cells expressing human CD19 (Ling, NR, et al. (1987). Leucocyte typing III. 302). The FMC63 antibody contains CDRH1 and H2 listed in SEQ ID NOS: 44, 45, respectively, and CDRH3 listed in SEQ ID NOS: 46 or 47, and CDRL1 listed in SEQ ID NOS: 48, and CDR L2 listed in SEQ ID NO: 49 or 50, and the CDR L3 sequences shown in SEQ ID NO: 51 or 52. The FMC63 antibody contains a heavy chain variable region (V _H ) containing the amino acid sequence of SEQ ID NO: 53 and a light chain variable region (V _L ) comprising the amino acid sequence of SEQ ID NO: 54. In some embodiments, the svFv comprises a variable light chain comprising the CDRL1 sequence of SEQ ID NO:48, the CDRL2 sequence of SEQ ID NO:49, and the CDRL3 sequence of SEQ ID NO:51 , and/or a heavy chain variable containing the sequence from CDRH1 of SEQ ID NO:44, the CDRH2 sequence from SEQ ID NO:45, and the sequence from CDRH3 of SEQ ID NO:46. In some embodiments, the scFv comprises the FMC63 heavy chain variable region set forth in SEQ ID NO:53 and the FMC63 light chain variable region set forth in SEQ ID NO:54. In some embodiments, the variable heavy and variable light chains are connected by a linker. In some embodiments, the linker is listed in SEQ ID NO: 70, 72, 73, 74, or 189. In some embodiments, the scFv contains, in that order, V _H , linker, and V _L . In some embodiments, the scFv contains, in that order, V _L , a linker, and V _H . In some embodiments, the svFv is encoded by the nucleotide sequence shown in SEQ ID NO:69 or by a sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92 %, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:69. In some embodiments, the scFv contains the amino acid sequence shown in SEQ ID NO:55, or a sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92 %, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:55.

[0392] In some embodiments, the implementation of the antigen-binding domain contains V _H and/or V _L derived from SJ25C1, which, in some aspects, may be scFv. SJ25C1 is a mouse IgG1 monoclonal antibody raised against Nalm-1 and -16 cells expressing human CD19 (Ling, NR, et al. (1987). Leucocyte typing III. 302). The SJ25C1 antibody contains the CDRH1, H2, and H3 sequences shown in SEQ ID NOS: 59-61, respectively, and the CDRL1, L2, and L3 sequences shown in SEQ ID NOS: 56-58, respectively. The SJ25C1 antibody comprises a heavy chain variable region (V _H ) containing the amino acid sequence from SEQ ID NO: 62 and a light chain variable region (V _L ) containing the amino acid sequence from SEQ ID NO: 63. In some embodiments, the svFv contains the variable a light chain containing a CDRL1 sequence from SEQ ID NO:56, a CDRL2 sequence from SEQ ID NO:57 and a sequence from CDRL3 of SEQ ID NO:58 and/or a variable heavy chain containing a CDRH1 sequence from SEQ ID NO:59, a CDRH2 sequence from SEQ ID NO:60 and sequence from CDRH3 SEQ ID NO:61. In some embodiments, the scFv comprises the SJ25C1 heavy chain variable region set forth in SEQ ID NO:62 and the SJ25C1 light chain variable region set forth in SEQ ID NO:63. In some embodiments, the variable heavy and variable light chains are connected by a linker. In some embodiments, the linker is shown in SEQ ID NO:64. In some embodiments, the scFv contains, in that order, V _H , a linker, and V _L . In some embodiments, the scFv contains, in that order, V _L , a linker, and V _H . In some embodiments, the scFv contains the amino acid sequence shown in SEQ ID NO:65 or a sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92 %, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO:65.

[0393] In some embodiments, the antibody is an antigen-binding fragment, such as scFv, that contains one or more linkers connecting two antibody domains or regions, such as a heavy chain variable region (V _H ) and a light chain variable region ( V _L ). Accordingly, antibodies comprise single chain antibody fragments, such as scFv, and diabodies, in particular human single chain antibody fragments, typically containing a linker(s) connecting two antibody domains or regions, such as the V _H and V _L regions. Typically, the linker is a peptide linker, eg a flexible and/or soluble peptide linker, eg enriched in glycine and serine. Among the linkers are enriched in glycine and serine, and/or, in some cases, threonine. In some embodiments, the linkers further contain charged moieties such as lysine and/or glutamate, which may improve solubility. In some embodiments, the linkers further comprise one or more prolines.

[0394] In some aspects, linkers enriched in glycine and serine (and/or threonine) contain at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of such amino acid(s). In some embodiments, they contain at least, or about at least 50%, 55%, 60%, 70%, or 75%, glycine, serine, and/or threonine. In some embodiments, the linker consists essentially entirely of glycine, serine, and/or threonine. Typically, linkers contain between about 5 and about 50 amino acids, typically between (or about) 10 and (or about) 30, e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30, and in some examples, between 10 and 25 amino acids. Exemplary linkers include linkers containing varying numbers of repeats of the sequence GGGGS (4GS; SEQ ID NO:19) or GGGS (3GS; SEQ ID NO:71), for example between 2, 3, 4 and 5 repeats of such a sequence. Illustrative linkers include linkers containing, or consisting of, the sequence shown in SEQ ID NO: 72 (GGGGSGGGGSGGGGS), SEQ ID NO: 189 (ASGGGGSGGRASGGGGS), SEQ ID NO: 73 (GSTSGSGKPGSGEGSTKG) or SEQ ID NO: 74 (SRGGGGSGGGGSGGGGSLEMA ).

[0395] In some embodiments, a recombinant receptor, such as CAR, such as part of an antibody of a recombinant receptor, such as CAR, further comprises a spacer, which may be or contain at least a portion of an immunoglobulin constant region, or a variant or modified thereof version, such as the hinge region, for example, the IgG4 hinge region, the IgG1 hinge region, C _H 1/C _L and/or the Fc region. In some embodiments, the recombinant receptor further comprises a spacer and/or a hinge region. In some embodiments, the constant region or portion is derived from human IgG, such as IgG4 or IgG1. In some aspects, a portion of the constant region serves as a spacer region between the antigen recognition component, eg, scFv, and the transmembrane domain. The spacer may be of a length that provides increased responsiveness of the cell after antigen binding compared to no spacer.

[0396] Exemplary spacers, such as hinge regions, comprise the regions described in International Patent Application Publication WO2014031687. In some examples, the spacer contains or approximately contains 12 amino acids, or contains no more than 12 amino acids. Exemplary spacers include spacers containing at least about 10-229 amino acids, about 10-200 amino acids, about 10-175 amino acids, about 10-150 amino acids, about 10-125 amino acids, about 10-100 amino acids, about 10- 75 amino acids, about 10-50 amino acids, about 10-40 amino acids, about 10-30 amino acids, about 10-20 amino acids, or about 10-15 amino acids, and including any integer between the endpoints of any of the listed ranges. In some embodiments, the spacer region is about 12 amino acids or less, about 119 amino acids or less, or about 229 amino acids or less. In some embodiments, the spacer is a spacer having at least a specific length, such as a length that is at least 100 amino acids, such as at least 110, 125, 130, 135, 140, 145 , 150, 160, 170, 180, 190, 200, 210, 220, 230, 240 or 250 amino acids. Exemplary spacers include an IgG4 hinge per se, an IgG4 hinge associated with C _H 2 and C _H 3 domains, or an IgG4 hinge associated with a C _H 3 domain. Exemplary spacers include an IgG4 hinge per se, an IgG4 hinge associated with C domains _H 2 and C _H 3, or an IgG4 hinge associated with a C _H 3 domain. Exemplary spacers include, but are not limited to, those described in Hudecek et al., Clin. Cancer Res., 19:3153 (2013), Hudecek et al. (2015) Cancer Immunol Res. 3(2): 125-135, in International Patent Application Publication No. WO2014031687, in US Patent No. 8822647 or in US Published Application No. 2014/0271635. In some embodiments, the spacer comprises an immunoglobulin hinge, C _H 2 and C _H 3 region sequence. In some embodiments, one or more of the C _H 2 and C _H 3 hinge groups are derived entirely or partially from IgG4 or IgG2. In some cases, the C _H 2 and C _H 3 hinges are derived from IgG4. In some aspects, one or more of the C _H 2 and C _H 3 hinge groups is chimeric and contains a sequence derived from IgG4 and IgG2. In some examples, the spacer contains an IgG4/2 chimeric hinge, an IgG2/4 C _H 2 region, and an IgG4 C _H 3 region.

[0397] In some embodiments, the spacer, which may be a constant region or part thereof, of an immunoglobulin is derived from a human IgG, such as IgG4 or IgG1. In some embodiments, the spacer contains the sequence ESKYGPPCPPCP (shown in SEQ ID NO: 1). In some embodiments, the spacer contains the sequence shown in SEQ ID NO: 3. In some embodiments, the spacer has the sequence shown in SEQ ID NO: 4. In some embodiments, the encoded spacer is or contains the sequence shown in SEQ ID NO: 29. In some embodiments, the constant region or portion is derived from IgD. In some embodiments, the spacer contains the sequence shown in SEQ ID NO: 5. In some embodiments, the spacer contains the sequence shown in SEQ ID NO: 125.

[0398] In some embodiments, the spacer may be derived in whole or in part from IgG4 and/or IgG2 and may contain mutations, such as one or more single amino acid mutations in one or more domains. In some examples, the amino acid modification is a substitution of proline (P) instead of serine (S) in the IgG4 hinge region. In some embodiments, the amino acid modification is a substitution of glutamine (Q) instead of asparagine (N) to reduce glycosylation heterogeneity, such as the N177Q mutation at position 177, in the C _H 2 region of the full-length Fc IgG4 sequence, or N176Q at position 176, in region C _H 2 full-length sequence of Fc IgG4.

[0399] Other exemplary spacer regions include hinge regions derived from CD8a, CD28, CTLA4, PD-1, or FcγRIIIa. In some embodiments, the spacer comprises a truncated extracellular domain or hinge region of CD8a, CD28, CTLA4, PD-1, or FcγRIIIa. In some embodiments, the spacer is a truncated hinge region of CD28. In some embodiments, a short oligo or polypeptide linker, e.g., a 2-10 amino acid linker, such as an alanine or alanine and arginine containing linker, e.g., an alanine triplet (AAA) or RAAA (SEQ ID NO: 180), is present. and forms a link between the scFv and the CAR spacer region. In some embodiments, the spacer contains the sequence shown in SEQ ID NO: 114. In some embodiments, the spacer contains the sequence shown in SEQ ID NO: 116. In some embodiments, the spacer contains the sequence shown in any SEQ ID NO: 117-119. In some embodiments, the spacer contains the sequence shown in SEQ ID NO: 120. In some embodiments, the spacer contains the sequence shown in SEQ ID NO: 122. In some embodiments, the spacer contains the sequence shown in SEQ ID NO: 124 .

[0400] In some embodiments, the spacer contains an amino acid sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to any of SEQ ID NOS: 1, 3, 4, 5, or 29, 114, 116, 117, 118, 119, 120, 122, 124 or 125.

[0401] This antigen recognition domain is typically associated with one or more intracellular signaling components, such as signaling components that mimic stimulation and/or activation by an antigen-receptor complex, such as the TCR complex, in the case of CAR, and/ or a signal through another receptor on the cell surface. Thus, in some embodiments, the antigen-binding component (eg, antibody) is associated with one or more transmembrane and intracellular signaling domains. In some embodiments, the transmembrane domain is fused to the extracellular domain. In one embodiment, a transmembrane domain is used that naturally associates with one of the domains in the receptor, such as CAR. In some cases, a transmembrane domain is selected or modified by amino acid substitution to eliminate the binding of such domains to transmembrane domains of the same or different surface membrane proteins to minimize interactions with other elements of the receptor complex.

[0402] The transmembrane domain, in some embodiments, is derived from either a natural or synthetic source. When the source is natural, the domain in some aspects is derived from any membrane-bound or transmembrane protein. Transmembrane regions include regions derived from (i.e., they contain at least their transmembrane region(s)) T-lymphocyte receptor alpha, beta or zeta chains, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD8a, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 (4-1BB), CD154, CTLA-4 or PD-1. Alternatively, in some embodiments, the transmembrane domain is synthetic. In some aspects, the synthetic transmembrane domain predominantly contains hydrophobic residues such as leucine and valine. In some aspects, a triplet of phenylalanine, tryptophan and valine will be located at each end of the synthetic transmembrane domain. In some embodiments, linking is via linkers, spacers, and/or transmembrane domain(s). Exemplary transmembrane domain sequences are or contain the sequences shown in SEQ ID NOs: 8, 115, 121, 123, 178, or 179.

[0403] Among the intracellular signaling domains are those that reproduce or approximately reproduce the signal through the natural antigen receptor, the signal through such a receptor in combination with a costimulatory receptor, and/or the signal through the costimulatory receptor itself. In some embodiments, a short oligo or polypeptide linker is present, e.g., a linker of 2-10 amino acids, e.g., containing glycines and serines, e.g., a glycine-serine doublet, and forms a link between the transmembrane domain and the CAR cytoplasmic signaling domain.

[0404] A receptor, such as a CAR, typically contains at least one or more intracellular signaling components. In some embodiments, the receptor comprises an intracellular component of the TCR complex, such as the CD3 TCR chain, which mediates T-lymphocyte stimulation and/or activation and cytotoxicity, such as the CD3 zeta chain. Thus, in some aspects, the antigen-binding portion is associated with one or more signaling modules of the cell. In some embodiments, cell signaling modules include a CD3 transmembrane domain, CD3 intracellular signaling domains, and/or other CD transmembrane domains. In some embodiments, the receptor, eg, CAR, further comprises a portion of one or more additional molecules, such as an Fc receptor γ, CD8, CD4, CD25, or CD16. For example, in some aspects, the CAR or other chimeric receptor comprises a chimeric molecule of CD3-zeta (CD3-ζ) or Fc receptor γ and CD8, CD4, CD25, or CD16.

[0405] In some embodiments, upon ligation of a CAR or other chimeric receptor, the cytoplasmic domain or intracellular signaling domain of the receptor stimulates and/or activates at least one of the normal effector functions or responses of an immune cell, such as a genetically engineered T- lymphocyte to express CAR. For example, in some contexts, CARs induce T-lymphocyte function, such as cytolytic activity or T-helper activity, such as the secretion of cytokines or other factors. In some embodiments, a truncated portion of the intracellular signaling domain of an antigen receptor component or costimulatory molecule is used in place of an intact immunostimulatory chain, for example if it transduces an effector function signal. In some embodiments, the intracellular signaling domain or domains comprise cytoplasmic T-lymphocyte receptor (TCR) sequences, and in some aspects also those co-receptors that naturally act in concert with such receptors to initiate signal transduction upon antigen engagement and receptor, and/or any derivative or variant of such molecules and/or any synthetic sequence that has the same functionality.

[0406] In the context of natural TCRs, complete activation typically requires not only signaling through the TCR, but also a costimulatory signal. Thus, in some embodiments, to facilitate full activation, a component for generating a secondary or costimulatory signal is also included in the CAR. In other embodiments, the CAR does not contain a component for generating a costimulatory signal. In some aspects, the additional CAR is expressed in the same cell and provides a component for generating a secondary or co-stimulatory signal.

[0407] Stimulation and/or activation of T-lymphocytes in some aspects is described as mediated by two classes of cytoplasmic signaling sequences: those that initiate primary antigen-dependent stimulation and/or activation through TCRs (primary cytoplasmic signaling regions, domains or sequences), and those that act in an antigen-independent manner to produce a secondary or costimulatory signal (secondary cytoplasmic signaling regions, domains or sequences). In some aspects, the CAR contains one or both of these signaling components.

[0408] In some aspects, the CAR contains primary cytoplasmic signaling regions, domains, or sequence that regulates the primary activation of the TCR complex. Primary cytoplasmic signaling regions, domains or sequences that act in a stimulatory manner may contain signaling motifs, which are known as immunoreceptor tyrosine activating motifs or ITAMs. Examples of ITAMs containing primary cytoplasmic signal sequences include groups derived from TCR zeta, FcR gamma, FcR beta, CD3 gamma, CD3 delta, CD3 epsilon, CD8, CD22, CD79a, CD79b, and CD66d. In some embodiments, the cytoplasmic signaling molecule(s) in the CAR contains (comprises) a cytoplasmic signaling domain, a portion thereof, or a sequence derived from CD3 zeta. In some embodiments, the CAR comprises the signaling region and/or the transmembrane portion of a costimulatory receptor such as CD28, 4-1BB, OX40 (CD134), CD27, DAP10, DAP12, ICOS, and/or other costimulatory receptors. In some aspects, the same CAR contains both a primary cytoplasmic signaling region and costimulatory signaling components.

[0409] In some embodiments, one or more different recombinant receptors may contain one or more different intracellular signaling regions or domains. In some embodiments, the primary cytoplasmic signaling region is included in the CAR while the costimulatory component is provided by another receptor, such as another CAR recognizing a different antigen. In some embodiments, CARs include activating or stimulatory CARs and costimulatory CARs, all of which are expressed on the same cell (see WO2014/055668).

[0410] In some aspects, the cells contain one or more stimulatory or activating CARs and/or costimulatory CARs. In some embodiments, cells further comprise inhibitory CARs (iCARs, see Fedorov et al., Sci. Transl. Medicine, 5(215) (2013), such as CARs recognizing an antigen other than an antigen associated and/or specific for disease or condition, wherein the activating signal delivered by the CAR to the disease is reduced or inhibited by binding the inhibitory CAR to its ligand, for example, to reduce off-target effects.

[0411] In certain embodiments, the intracellular signaling domain comprises a CD28 transmembrane and signaling domain associated with an intracellular CD3 domain (eg, CD3-zeta). In some embodiments, the intracellular signaling domain comprises chimeric CD28 and CD137 (4-1BB, TNFRSF9) co-stimulatory domains associated with the CD3 zeta intracellular domain.

[0412] In some embodiments, the CAR encompasses one or more, eg, two or more, costimulatory domains and a primary cytoplasmic signaling region in the cytoplasmic portion. Exemplary CARs include intracellular components such as the intracellular signaling region(s) or domain(s) of CD3-zeta, CD28, CD137 (4-1BB), OX40 (CD134), CD27, DAP10, DAP12, NKG2D, and/or ICOS. In some embodiments, the chimeric antigen receptor comprises an intracellular signaling region or co-stimulatory T-lymphocyte molecule domain, e.g. in some cases, between a transmembrane domain and an intracellular signaling region or domain. In some aspects, the T lymphocyte costimulatory molecule is one or more of CD28, CD137 (4-1BB), OX40 (CD134), CD27, DAP10, DAP12, NKG2D and/or ICOS.

[0413]

[0414] In some cases, CARs are referred to as first, second, and/or third generation CARs. In some aspects, a first generation CAR is a CAR that provides only the signal induced by the CD3 chain upon antigen binding; in some aspects, a second generation CAR is a CAR that provides such a signal and a costimulatory signal, eg, a signal comprising an intracellular signaling domain from a costimulatory receptor such as CD28 or CD137; in some aspects, a third generation CAR is a CAR that contains multiple co-stimulatory domains of various co-stimulatory receptors.

[0415] In some embodiments, the chimeric antigen receptor comprises an extracellular portion containing an antibody or antibody fragment. In some aspects, the chimeric antigen receptor contains an extracellular portion containing the antibody or fragment, and an intracellular signaling domain. In some embodiments, the antibody or fragment contains scFv and the intracellular domain contains ITAM. In some aspects, the intracellular signaling domain comprises a zeta chain, CD3-zeta (CD3ζ) chain signaling domain. In some embodiments, the chimeric antigen receptor comprises a transmembrane domain linking an extracellular domain and an intracellular signaling domain. In some aspects, the transmembrane domain contains the transmembrane portion of CD28. In some embodiments, the chimeric antigen receptor comprises the intracellular domain of a costimulatory T-lymphocyte molecule. The extracellular domain and the transmembrane domain can be linked directly or indirectly. In some embodiments, the extracellular domain and the transmembrane domain are linked by a spacer, such as any spacer described herein. In some embodiments, the receptor comprises an extracellular portion of the molecule from which the transmembrane domain is derived, such as the extracellular portion of CD28. In some embodiments, the chimeric antigen receptor comprises an intracellular domain derived from a co-stimulatory T-lymphocyte molecule or a functional variant thereof, such as a link between the transmembrane domain and the intracellular signaling domain. In some aspects, the T-lymphocyte costimulatory molecule is CD28 or 4-1BB.

[0416] For example, in some embodiments, the CAR comprises an antibody, e.g., an antibody fragment, a transmembrane domain that is or contains the transmembrane portion of CD28 or a functional variant thereof, and an intracellular signaling domain comprising the signal portion of CD28 or a functional variant thereof and a signal part of CD3 zeta or its functional variant. In some embodiments, the CAR comprises an antibody, e.g., an antibody fragment, a transmembrane domain that is or contains a CD28 transmembrane portion or a functional variant thereof, and an intracellular signaling domain comprising a 4-1BB signaling portion or a functional variant thereof and a CD3 zeta signaling portion. or its functional variant. In some such embodiments, the receptor further comprises a spacer comprising a portion of an Ig molecule, such as a human Ig molecule, such as an Ig hinge, eg, an IgG4 hinge, such as a hinge-only spacer.

[0417] In some embodiments, the transmembrane domain of the recombinant receptor, e.g., CAR, is or comprises a transmembrane domain of human CD28 (e.g., Accession No. P10747.1) or CD8a (Accession No. P01732.1), or a variant thereof, such as a transmembrane domain that contains the amino acid sequence shown in SEQ ID NO: 8, 115, 178, or 179, or an amino acid sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90 %, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with SEQ ID NO: 8, 115, 178 or 179; in some embodiments, the transmembrane domain-containing portion of the recombinant receptor comprises the amino acid sequence shown in SEQ ID NO: 9, or an amino acid sequence having at least, or about, at least 85%, 86%, 87% , 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with it.

[0418] In some embodiments, the recombinant receptor intracellular signaling component(s), e.g., CAR, comprises the human CD28 intracellular co-stimulatory signaling domain, or a functional variant or portion thereof, such as the GG LL substitution domain at positions 186-187 of the native CD28 protein . For example, an intracellular signaling domain may contain the amino acid sequence shown in SEQ ID NO: 10 or 11, or an amino acid sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91% , 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with SEQ ID NO: 10 or 11. In some embodiments, the intracellular domain comprises an intracellular costimulatory signaling domain 4 -1BB (e.g., Accession No. Q07011.1), or a functional variant or portion thereof, such as the amino acid sequence shown in SEQ ID NO: 12, or an amino acid sequence that exhibits at least 85%, 86%, 87 %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with SEQ ID NO: 12.

[0419] In some embodiments, the intracellular signaling domain of the recombinant receptor, e.g., CAR, comprises a human CD3 zeta stimulatory signaling domain or a functional variant thereof, such as the human CD3ζ isoform 3 cytoplasmic domain 112 AA (Accession No. P20963,2) or signaling a CD3 zeta domain as described in US Pat. demonstrates at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity with SEQ ID NO: 13, 14 or 15.

[0420] In some aspects, the spacer contains only an IgG hinge region, such as an IgG4 or IgG1 hinge region per se, such as a hinge-only spacer set forth in SEQ ID NO: 1 or SEQ ID NO: 125. In other embodiments, implementation, the spacer is or contains an Ig hinge region, for example, a hinge region derived from IgG4, optionally associated with CH2 and/or CH3 domains. In some embodiments, the spacer is an Ig hinge, e.g., an IgG4 hinge associated with the CH2 and CH3 domains as set forth in SEQ ID NO: 4. In some embodiments, the spacer is an Ig hinge, e.g. IgG4 bound only to the CH3 domain as set forth in SEQ ID NO: 3. In some embodiments, the spacer is or contains a glycine-serine enriched sequence or other flexible linker such as known flexible linkers. In some embodiments, the spacer is a CD8a hinge such as given in any of SEQ ID NOs: 117-119, an FcγRIIIa hinge as given in SEQ ID NO: 124, a CTLA4 hinge as given in SEQ ID NOs: 120, or a hinge region PD-1 such as shown in SEQ ID NO: 122.

[0421] For example, in some embodiments, the CAR comprises an antibody, such as an antibody fragment, including scFv, a spacer, such as a spacer containing a portion of an immunoglobulin molecule, such as a hinge region, and/or one or more constant regions of a heavy chain molecule , such as a spacer containing an Ig hinge region, a transmembrane domain containing all or part of the transmembrane domain derived from CD28, an intracellular signaling domain derived from CD28, and a CD3 zeta signaling domain. In some embodiments, the CAR comprises an antibody or fragment such as an scFv, a spacer such as any of the spacers comprising an Ig hinge region, a CD28-derived transmembrane domain, a 4-1BB-derived intracellular signaling domain, and a CD3-derived zeta signaling domain. .

[0422] Recombinant receptors, such as CARs, expressed by cells administered to a subject typically recognize or specifically bind the molecule that is expressed, in the disease or condition, or their cells being treated, associates with, and/or is specific to them . When specifically bound to a molecule, such as an antigen, the receptor typically delivers an immunostimulatory signal, such as an ITAM-transduced signal, into the cell, thereby facilitating an immune response targeting the disease or condition. For example, in some embodiments, the cells express a CAR that specifically binds to an antigen expressed by, or associated with, the disease or condition, cell or tissue. Non-limiting exemplary CAR sequences are shown in SEQ ID NOs: 126-177.

[0423] In some embodiments, the encoded CAR sequence may further comprise a signal sequence or signal peptide that directs or delivers the CAR to the cell surface in which the CAR is expressed. In some embodiments, the signal peptide is derived from a transmembrane protein. In some examples, the signal peptide is derived from CD8a, CD33, or IgG. Illustrative signal peptides include the sequences shown in SEQ ID NOs: 21, 75 and 76, or a variant thereof.

[0424] In some embodiments, the CAR comprises an anti-CD19 antibody, such as an antibody fragment, including scFv, a spacer, such as a spacer containing an immunoglobulin moiety, such as a hinge region, and/or one or more constant regions of a heavy chain molecule , such as spacers containing any Ig hinge regions or other spacers described herein, a transmembrane domain containing all or part of it CD28 transmembrane domain derived, CD28 derived intracellular signaling domain and CD3 zeta signaling domain. In some embodiments, the CAR comprises an anti-CD19 antibody or fragment such as an scFv, a spacer such as any of the spacers containing an Ig hinge region, or other spacers described herein, a CD28-derived transmembrane domain derived from 4 -1BB intracellular signaling domain and CD3-derived zeta signaling domain. In some embodiments, such CAR constructs further comprise a T2A ribosome jumper and/or a tEGFR sequence, eg, downstream of the CAR.

[0425] In some embodiments, the CAR comprises an anti-BCMA antibody or fragment, such as any of the human anti-BCMA antibodies, including sdAbs and scFv described herein, a spacer, such as any of the spacers containing the Ig hinge region , or other spacers described herein, CD28 transmembrane domain, CD28 intracellular signaling domain, and CD3 zeta signaling domain. In some embodiments, the CAR comprises an anti-BCMA antibody or fragment, such as any of the human anti-BCMA antibodies, including the sdAbs and scFv described herein, a spacer, such as any of the spacers containing an Ig hinge region, or others. the spacers described herein, the CD28 transmembrane domain, the 4-1BB intracellular signaling domain, and the CD3 zeta signaling domain. In some embodiments, such CAR constructs further comprise a T2A ribosome jumper and/or a tEGFR sequence, eg, downstream of the CAR.

2. Chimeric Autoantibody Receptor (CAAR)

[0426] In some embodiments, the recombinant receptor is a chimeric autoantibody receptor (CAAR). In some embodiments, the CAAR binds, eg, specifically binds to, or recognizes an autoantibody. In some embodiments, cells expressing CAAR, such as engineered T lymphocytes for expressing CAAR, can be used to bind and kill autoantibody expressing cells, but not cells expressing normal antibodies. In some embodiments, CAAR-expressing cells can be used to treat an autoimmune disease associated with the expression of autoantigens, such as autoimmune diseases. In some embodiments, CAAR-expressing cells can target B lymphocytes that eventually produce autoantibodies and display autoantibodies on their cell surfaces, label these B lymphocytes as disease-specific targets for therapeutic intervention. In some embodiments, CAAR-expressing cells can be used to effectively target and kill pathogenic B lymphocytes in autoimmune diseases by targeting disease-causing B lymphocytes using an antigen-specific chimeric autoantibody receptor. In some embodiments, the recombinant receptor is a CAAR, such as any CAAR described in US Patent Application Publication No. 2017/0051035.

[0427] In some embodiments, the CAAR comprises an autoantibody-binding domain, a transmembrane domain, and one or more intracellular signaling regions or domains (also referred to interchangeably as a cytoplasmic signaling domain or region). In some embodiments, the intracellular signaling region contains an intracellular signaling domain. In some embodiments, the intracellular signaling domain is or contains a primary signaling region, a signaling domain that can stimulate and/or induce a primary activation signal in a T-lymphocyte, a T-lymphocyte receptor (TCR) component signaling domain (e.g., an intracellular signaling domain or a CD3-zeta chain (CD3ζ) chain zeta region or a functional variant thereof or a signal portion thereof), and/or an immunoreceptor tyrosine activating motif (ITAM) signaling domain.

[0428] In some embodiments, the autoantibody-binding domain comprises a self-antigen or fragment thereof. The choice of autoantigen may depend on the type of autoantibody targeted. For example, the autoantigen may be selected because it recognizes an autoantibody on a target cell, such as a B-lymphocyte, associated with a particular disease state, such as an autoimmune disease, such as an autoantibody-mediated autoimmune disease. In some embodiments, the autoimmune disease includes pemphigus vulgaris (PV). Exemplary autoantigens include desmoglein 1 (Dsg1) and Dsg3.

3.TCR

[0429] In some embodiments, engineered cells, such as T-lymphocytes, are provided that express a T-lymphocyte receptor (TCR), or an antigen-binding portion thereof, that recognizes a peptide or T-lymphocyte epitope of a target polypeptide, such as an antigen. tumors, viral or autoimmune protein. In some aspects, the TCR is or contains a recombinant TCR.

[0430] In some embodiments, a "T-lymphocyte receptor" or "TCR" is a molecule that contains α and β variable chains (also known as TCRα and TCRβ, respectively) or γ and δ variable chains (also known as TCRγ and TCRδ, respectively) or antigen-binding portions thereof, and which are capable of specifically binding to a peptide associated with an MHC molecule. In some embodiments, the TCR is in the αβ form. Typically, TCRs that are in the αβ and γδ forms are broadly structurally similar, but T lymphocytes expressing them may have different anatomical positions or functions. The TCR may be on the surface of the cell or in a soluble form. Typically, the TCR is found on the surface of T-lymphocytes (or T-lymphocytes), where they are generally responsible for recognizing antigens associated with major histocompatibility complex (MHC) molecules.

[0431] Unless otherwise stated, the term “TCR” is understood to encompass full length TCRs as well as their antigen-binding portions or antigen-binding fragments. In some embodiments, the TCR is an intact or full length TCR, including TCR in the αβ form or in the γδ form. In some embodiments, the TCR is an antigen-binding portion that is smaller than the full length TCR, but that binds to a specific peptide bond in the MHC molecule, eg, binds to an MHC-peptide complex. In some cases, an antigen-binding portion or fragment of a TCR may contain only a portion of the structural domains of a full-length or intact TCR, but in addition it is capable of binding a peptide epitope, such as an MHC-peptide complex, to which the full-length TCR binds. In some cases, the antigen-binding portion contains TCR variable domains, such as the TCR variable α and β variable chain, sufficient to form a binding site for binding a particular MHC-peptide complex. Typically, TCR variable chains contain complementarity defining regions involved in recognition of the peptide, MHC, and/or MHC-peptide complex.

[0432] In some embodiments, the TCR variable domains comprise hypervariable loops or complementarity determining regions (CDRs), which typically make a major contribution to antigen recognition and binding abilities and specificity. In some embodiments, the TCR CDR, or a combination thereof, forms all or substantially all of the antigen-binding site of a given TCR molecule. The various CDRs in the variable region of the TCR chain are generally separated by frame regions (FRs), which tend to exhibit less variability among TCR molecules compared to CDRs (see, e.g., Jores et al., Proc. Nat'l Acad. Sci U.S.A. 87:9138, 1990; Chothia et al., EMBO J. 7:3745, 1988; see also Lefranc et al., Dev. Comp. Immunol. 27:55, 2003). In some embodiments, the CDR3 is the major CDR responsible for antigen binding or specificity, or is the most important of the three CDRs in a given TCR variable region for antigen recognition and/or interaction with the tested peptide portion of the peptide-MHC complex. In some contexts, the CDR1 alpha chain may interact with the N-terminus of certain antigenic peptides. In some contexts, the beta chain CDR1 may interact with the C-terminus of the peptide. In some contexts, CDR2 is the main contributor or is the main CDR responsible for interacting with or recognizing the MHC portion of the MHC-peptide complex. In some embodiments, the β-chain variable region may contain an additional hypervariable region (CDR4 or HVR4) that is typically involved in superantigen binding rather than antigen recognition (Kotb (1995) Clinical Microbiology Reviews, 8:411-426 ).

[0433] In some embodiments, the TCR may also comprise a constant domain, a transmembrane domain, and/or a short cytoplasmic tail (see, e.g., Janeway et al., Immunobiology: The Immune System in Health and Disease, 3rd Ed., Current Biology Publications , p. 4:33, 1997). In some aspects, each TCR chain may have one N-terminal immunoglobulin variable domain, one immunoglobulin constant domain, a transmembrane region, and a short cytoplasmic tail at the C-terminal edge. In some embodiments, the TCR is associated with invariant CD3 complex proteins involved in signal transduction mediation.

[0434] In some embodiments, the TCR chain contains one or more constant domains. For example, the extracellular portion of a given TCR chain (eg, α-chain or β-chain) may contain two immunoglobulin-like domains, such as a variable domain (eg, Vα or Vβ; typically amino acids 1-116 based on Kabat, Kabat numbering). et al., “Sequences of Proteins of Immunological Interest, US Dept. Health and Human Services, Public Health Service National Institutes of Health, 1991, 5th ed.) and a constant domain (e.g., α-chain constant domain or Cα, usually , chain positions 117-259 based on Kabat numbering, or chain constant domain β or Cβ, typically chain positions 117-295 according to Kabat) near the cell membrane. For example, in some cases, the extracellular part of the TCR, formed from two chains, contains two constant domains remote from the membrane and two variable domains close to the membrane, each of these variable domains contains a CDR. The TCR constant domain may contain short junction sequences in which a cysteine residue forms a disulfide bond, thereby joining two TCR chains. In some embodiments, the TCR may contain an additional cysteine residue in each of the α and β chains such that the TCR contains two disulfide bonds in the constant domains.

[0435] In some embodiments, the TCR chains contain a transmembrane domain. In some embodiments, the transmembrane domain is positively charged. In some cases, the TCR chain contains a cytoplasmic tail. In some cases, the structure allows the TCR to associate with other CD3-like molecules and their subunits. For example, a TCR containing constant domains with a transmembrane region can fix a protein in the cell membrane and associate with invariant signaling subunits or the CD3 complex. The intracellular tails of the CD3 signaling subunits (eg, CD3γ, CD3δ, CD3ε, and CD3ζ chains) contain one or more immunoreceptor tyrosine activating motifs or ITAMs that are involved in the signaling capability of the TCR complex.

[0436] In some embodiments, the TCR may be a two-chain heterodimer of α and β (or optionally γ and δ), or it may be a single chain TCR construct. In some embodiments, the TCR is a heterodimer containing two separate chains (α and β chains or γ and δ chains) that are linked, for example, by a disulfide bond or disulfide bonds.

[0437] In some embodiments, the TCR may be generated from known TCR sequence(s), such as the Vα,β chain sequences, for which a substantially full length coding sequence is readily available. Methods for obtaining full-length TCR sequences, including V chain sequences, from cellular sources are well known. In some embodiments, TCR-encoding nucleic acids can be obtained from a variety of sources, such as polymerase chain reaction (PCR), amplification of TCR-encoding nucleic acids within or isolated from a given cell or cells, or synthesis of publicly available TCR DNA sequences. .

[0438] In some embodiments, the recombinant receptors include recombinant TCRs and/or TCRs cloned from naturally occurring T lymphocytes. In some embodiments, a T lymphocyte clone with high affinity for a target antigen (eg, a cancer antigen) is identified, isolated from a patient, and introduced into cells. In some embodiments, a TCR clone for the target antigen is generated in genetically engineered transgenic mice with genes for the human immune system (eg, human leukocyte antigen or HLA system). See, for example, tumor antigens (see, for example, Parkhurst et al. (2009) Clin Cancer Res. 15:169-180 and Cohen et al. (2005) J Immunol. 175:5799-5808. In some embodiments, use phage display for isolating TCRs against a target antigen (see e.g. Varela-Rohena et al. (2008) Nat Med. 14:1390-1395 and Li (2005) Nat Biotechnol. 23:349-354.

[0439] In some embodiments, the TCR is derived from a biological source, eg, cells, eg, T-lymphocytes (eg, cytotoxic T-lymphocytes), T-lymphocyte hybridomas, or other publicly available source. In some embodiments, T-lymphocytes can be obtained from isolated cells in vivo. In some embodiments, the TCR is thymically selected TCR. In some embodiments, the TCR is a neoepitope-restricted TCR. In some embodiments, the T lymphocytes may be a cultured hybridoma or a clone of T lymphocytes. In some embodiments, the TCR, or an antigen-binding portion thereof, can be generated synthetically from knowledge of the sequence of the TCR.

[0440] In some embodiments, the TCR is generated from a TCR identified or selected from a TCR candidate library screened against a target polypeptide antigen or T-lymphocyte target epitope thereof. TCR libraries can be generated by amplifying the Vα and Vβ repertoire from T lymphocytes isolated from subjects, including cells present in the PBMC, spleen, or other lymphoid organ. In some cases, T-lymphocytes can be amplified from antitumor effector lymphocytes (TILs). In some embodiments, TCR libraries can be generated from CD4+ or CD8+ cells. In some embodiments, the TCR can be amplified from a normal healthy subject's T-lymphocyte source, ie, normal TCR libraries. In some embodiments, the TCR can be amplified from the diseased subject's T-lymphocyte source, ie, diseased TCR libraries. In some embodiments, degenerative primers are used to amplify the Vα and Vβ gene repertoire, for example by RT-PCR, in samples such as human T lymphocytes. In some embodiments, scTv libraries may be assembled from libraries naive Vα and Vβ, in which the amplified products are cloned or assembled for linker separation. Depending on the source of the subject and the cells, the libraries may be specific for the HLA allele. Alternatively, in some embodiments, TCR libraries may be generated by mutagenesis or diversification of the parent or framework TCR molecule. In some aspects, the TCR is subjected to directed evolution, eg via mutagenesis, eg, α or β chain. In some aspects, specific residues in the CDR of the TCR are changed. In some embodiments, the selected TCRs can be modified through affinity maturation. In some embodiments, antigen-specific T lymphocytes can be selected, for example by screening, to evaluate CTL activity against a peptide. In some aspects, TCRs, eg, those present on antigen-specific T lymphocytes, may be selected, eg, for binding activity, eg, for specific affinity or avidity for an antigen.

[0441] In some embodiments, the TCR, or an antigen-binding portion thereof, is a group that is modified or that is genetically engineered. In some embodiments, guided evolution techniques are used to generate TCRs with altered properties, such as higher affinity for a particular MHC-peptide complex. In some embodiments, directed evolution is achieved using display methods, including but not limited to yeast display (Holler et al., (2003) Nat Immunol, 4, 55-62; Holler et al., (2000) Proc Natl Acad Sci U S A, 97, 5387-92), phage display (Li et al., (2005) Nat Biotechnol, 23, 349-54), or T-lymphocyte display (Chervin et al., (2008) J Immunol Methods , 339, 175-84). In some embodiments, display approaches include genetic engineering or modification of a known parent or reference TCR. For example, in some cases, wild-type TCRs can be used as a template to produce mutagenized TCRs in which the CDRs are mutated at one or more residues and mutants with a desired altered property, such as higher affinity for the desired target antigen, are selected.

[0442] In some embodiments, the peptides of the target polypeptide for use in the production or generation of the TCR of interest are known or can be readily identified in a routine manner. In some embodiments, peptides useful in generating TCRs or antigen-binding portions thereof can be determined based on the presence of an HLA restricted motif in a target polypeptide of interest, such as the target polypeptide described below. In some embodiments, peptides are identified using computer predictive models. in a routine way. In some embodiments, for predicting class I MHC binding sites, such models include, but are not limited to, ProPred1 (Singh and Raghava (2001) Bioinformatics 17(12):1236-1237), and SYFPEITHI (see Schuler et al. , (2007) Immunoinformatics Methods in Molecular Biology, 409(1): 75-93 2007. In some embodiments, the MHC restricted epitope is HLA-A0201, which is expressed in approximately 39-46% of all Europeans, and therefore , is a usable choice of MHC antigen for use in the preparation of a TCR or other MHC-peptide binding molecule.

[0443] HLA-A0201 binding motifs and cleavage sites for proteasomes and immune proteasomes are known using computer predictive models. For prediction of MHC class I binding sites, such models include, but are not limited to, ProPred1 (described in more detail in Singh and Raghava, ProPred: prediction of HLA-DR binding sites. BIOINFORMATICS 17(12):1236-1237 2001), and SYFPEITHI (see Schuler et al., SYFPEITHI, Database for Searching and T-Cell Epitope Prediction. in Immunoinformatics Methods in Molecular Biology, vol 409(1): 75-93 2007).

[0444] In some embodiments, the TCR, or an antigen-binding portion thereof, may be a recombinantly produced natural protein or a mutated form thereof in which one or more properties, such as binding characteristics, are altered. In some embodiments, the TCR may be obtained from one of a variety of animal species such as humans, mice, rats, or other mammals. The TCR may be associated with the cell or be in a soluble form. In some embodiments, for the purposes of the proposed methods, the TCR is in a cell-associated form expressed on the cell surface.

[0445] In some embodiments, the TCR is a full length TCR. In some embodiments, the TCR is an antigen-binding portion. In some embodiments, the TCR is a TCR dimer (dTCR). In some embodiments, the TCR is a single chain TCR (sc-TCR). In some embodiments, the dTCR or scTCR has the structures described in WO 03/020763, WO 04/033685, WO2011/044186.

[0446] In some embodiments, the TCR contains a sequence corresponding to a transmembrane sequence. In some embodiments, the TCR contains a sequence corresponding to cytoplasmic sequences. In some embodiments, the TCR may complex the TCR with CD3. In some embodiments, any of the TCRs, including dTCR or scTCR, can bind to signaling domains that give rise to active TCR on the surface of T lymphocytes. In some embodiments, the TCR is expressed on the surface of cells.

[0447] In some embodiments, the dTCR comprises a first polypeptide, wherein the sequence corresponding to the TCR α chain variable region sequence is fused to the N-terminus of the sequence corresponding to the TCR α chain extracellular constant region sequence, and a second polypeptide, where the sequence corresponding to the TCR variable region sequence chain β TCR, fused to the N-terminus of the sequence corresponding to the extracellular sequence of the constant region of the β TCR chain, these first and second polypeptides are linked by a disulfide bond. In some embodiments, the bond may correspond to a native interchain disulfide bond present in native αβ dimeric TCRs. In some embodiments, interchain disulfide bonds are not present in native TCRs. For example, in some embodiments, one or more cysteines can be introduced into the extracellular constant region sequences of a dTCR polypeptide pair. In some cases, both native and non-native disulfide bonds may be desirable. In some embodiments, the TCR contains a transmembrane sequence for membrane fixation.

[0448] In some embodiments, the dTCR comprises an α TCR chain comprising an α variable domain, an α constant domain, and a first dimerization motif, attached to the C-terminus of the α constant domain, and a β TCR chain containing the β variable domain, the β constant domain and the first dimerization motif attached to the C-terminus of the β constant domain, where the first and second dimerization motifs easily interact to form a covalent bond between the amino acid in the first dimerization motif and an amino acid in the second dimerization motif, linking together the α TCR chain and the β TCR chain.

C. и

A., J. Mol. Biol. 242, 655 (1994); Kurucz, I. et al., PNAS (USA) 90 3830 (1993); опубликованные Международные заявки PCT №№ WO 96/13593, WO 96/18105, WO99/60120, WO99/18129, WO 03/020763, WO2011/044186; и Schlueter, C. J. et al., J. Mol. Biol. 256, 859 (1996). В некоторых вариантах осуществления, scTCR содержит введенную ненативную дисульфидную связь между цепями для облегчения ассоциации цепей TCR (смотри, например, опубликованную Международную заявку PCT № WO 03/020763). В некоторых вариантах осуществления, scTCR представляет собой не имеющий дисульфидной связи усеченный TCR, в котором гетерологичные лейциновые молнии, слитые с его C-окончаниями, облегчают ассоциацию цепей (смотри, например, опубликованную Международную заявку PCT № WO99/60120). В некоторых вариантах осуществления, scTCR содержит вариабельный домен TCRα, ковалентно связанный с вариабельным доменом TCR β с помощью пептидного линкера (смотри, например, опубликованную Международную PCT № WO99/18129).[0449] In some embodiments, the TCR is an scTCR. Typically, scTCR can be generated using appropriate known methods, see, for example, Soo Hoo, W. F. et al., PNAS (USA) 89, 4759 (1992);

C. and

A., J. Mol. Biol. 242, 655 (1994); Kurucz, I. et al., PNAS (USA) 90 3830 (1993); published PCT International Applications Nos. WO 96/13593, WO 96/18105, WO99/60120, WO99/18129, WO 03/020763, WO2011/044186; and Schlueter, C. J. et al., J. Mol. Biol. 256, 859 (1996). In some embodiments, the scTCR contains an introduced non-native strand disulfide bond to facilitate association of the TCR strands (see, for example, PCT Publication No. WO 03/020763). In some embodiments, the scTCR is a disulfide-free truncated TCR in which heterologous leucine zippers fused to its C-terminals facilitate chain association (see, for example, PCT Publication No. WO99/60120). In some embodiments, the scTCR comprises a TCRα variable domain covalently linked to a TCR β variable domain via a peptide linker (see, for example, PCT International Publication No. WO99/18129).

[0450] In some embodiments, the scTCR comprises a first segment consisting of an amino acid sequence corresponding to the TCR α chain variable region, a second segment consisting of an amino acid sequence corresponding to the TCR β chain variable region sequence fused to the N-terminus of the amino acid sequence corresponding to sequences of the extracellular constant domain of the β TCR chain, and a linker sequence connecting the C-terminus of the first segment to the N-terminus of the second segment.

[0451] In some embodiments, the scTCR comprises a first segment consisting of an α chain variable region sequence fused to the N-terminus of an α extracellular constant chain domain sequence and a second segment consisting of a β chain variable region sequence fused to the N-terminus of the sequence an extracellular constant domain of the β chain; and a transmembrane domain sequence; and, optionally, a linker sequence linking the C-terminus of the first segment to the N-terminus of the second segment.

[0452] In some embodiments, the scTCR comprises a first segment consisting of a TCR β chain variable region sequence fused to the N-terminus of a β chain extracellular constant domain sequence, and a second segment consisting of an α chain variable region sequence fused to N- the end of an α chain extracellular constant domain sequence, and a transmembrane domain sequence, and optionally a linker sequence linking the C-terminus of the first segment to the N-terminus of the second segment.

[0453] In some embodiments, the scTCR linker that binds the first and second TCR segments can be any linker capable of forming a single polypeptide strand while retaining TCR binding specificity. In some embodiments, the linker sequence may, for example, have the formula -P-AA-P-, where P is proline and AA is an amino acid sequence, where the amino acids are glycine and serine. In some embodiments, the first and second segments are paired such that their variable region sequences are oriented for such binding. Therefore, in some cases, the linker is long enough to increase the distance between the C-terminus of the first segment and the N-terminus of the second segment, or vice versa, but not too long to block or reduce binding of the scTCR to the target ligand. In some embodiments, the linker may contain from or about 10 to 45 amino acids, such as 10-30 amino acids or 26-41 amino acid residues, such as 29, 30, 31, or 32 amino acids. In some embodiments, the linker has the formula -PGGG-(SGGGG)5-P-, where P is proline, G is glycine, and S is serine (SEQ ID NO: 16). In some embodiments, the linker has the sequence GSADDAKKDAAKKDGKS (SEQ ID NO: 17)

[0454] In some embodiments, the scTCR comprises a covalent disulfide bond linking a remainder of the α chain constant domain immunoglobulin region to a remainder of the β chain constant domain immunoglobulin region. In some embodiments, there is no disulfide bond between chains in native TCR. For example, in some embodiments, one or more cysteines can be introduced into the extracellular constant region sequences of the first and second segments of the scTCR polypeptide. In some cases, both native and non-native disulfide bonds may be desirable.

[0455] In some embodiments, the implementation of dTCR or scTCR containing introduced disulfide bonds between the chains, native disulfide bonds are not present. In some embodiments, one or more native cysteines forming native interchain disulfide bonds are replaced by another residue such as serine or alanine. In some embodiments, the introduced disulfide bond may be formed by mutating non-cysteine residues on the first and second segments in the cysteine. Exemplary non-native TCR disulfide bonds are described in PCT Published International Application No. WO2006/000830.

[0456] In some embodiments, the TCR, or an antigen-binding fragment thereof, exhibits an affinity for an equilibrium binding constant of the target antigen between or about 10 ^-5 and 10 ^-12 M, and including all individual values and ranges within those limits. In some embodiments, the target antigen is an MHC-peptide complex or ligand.

[0457] In some embodiments, the TCR-encoding nucleic acid or nucleic acids, such as the α and β chains, can be amplified by PCR, cloning, or other appropriate means and cloned into the appropriate expression vector or vectors. The expression vector may be any suitable recombinant expression vector and may be used to transform or transfect any suitable host. Suitable vectors include vectors designed for growth and propagation, or for expression, or both, such as plasmids and viruses.

[0458] In some embodiments, the vector may be pUC series (Fermentas Life Sciences), pBluescript series (Stratagene, La Jolla, Calif.), pET series (Novagen, Madison, Wis.), pGEX series (Pharmacia Biotech, Uppsala, Sweden) or the pEX series (Clontech, Palo Alto, Calif.). In some cases, bacteriophage vectors such as λG10, λGT11, λZapII (Stratagene), λEMBL4 and λNM1149 can also be used. In some embodiments, plant expression vectors can be used and include pBI01, pBI101.2, pBI101.3, pBI121, and pBIN19 (Clontech). In some embodiments, animal expression vectors include pEUK-Cl, pMAM, and pMAMneo (Clontech). In some embodiments, a viral vector is used, such as a retroviral vector.

[0459] In some embodiments, recombinant expression vectors can be prepared using standard recombinant DNA techniques. In some embodiments, the vectors may contain regulatory sequences, such as transcription and translation initiation codons and termination codes, that are specific to the type of host (e.g., bacteria, fungus, plant, or animal) to which the vector is to be administered, as appropriate, and given whether the vector is DNA based or RNA based. In some embodiments, the vector may contain a non-native promoter operably linked to a nucleotide sequence encoding a TCR or an antigen binding portion (or other MHC peptide binding molecule). In some embodiments, the promoter may be a non-viral promoter or a viral promoter such as the cytomegalovirus (CMV) promoter, the SV40 promoter, the RSV promoter, and a promoter found in the long terminal repeat of a mouse stem cell virus. Other known promoters are also contemplated.

[0460] In some embodiments, to generate a vector encoding a TCR, the α and β chains are PCR amplified with total cDNA isolated from a T lymphocyte clone expressing the TCR of interest and cloned into an expression vector. In some embodiments, the α and β chains are cloned into the same vector. In some embodiments, the α and β chains are cloned into different vectors. In some embodiments, the generated α and β chains are introduced into a retroviral, eg, lentiviral, vector.

4. Multitargeting

[0461] In some embodiments, cells and methods include multi-targeting strategies, such as expressing two or more engineered receptors on a cell, each recognizing the same or a different antigen, and typically each containing a different intracellular signaling component. Such multi-targeting strategies are described, for example, in PCT Publication No. WO 2014055668 A1 (describing combinations of activating and costimulatory CARs, e.g., targeting two different antigens that are present separately on non-targeted, e.g., normal cells, but present together only on disease or condition cells, to be treated), and Fedorov et al., Sci. Transl. Medicine, 5(215) (2013) (describes cells expressing activating and inhibitory CARs, such as those in which the activating CAR binds to a single antigen expressed both on normal or non-diseased cells and on cells of a disease or condition that should be treated, and the inhibitory CAR binds to another antigen expressed only on normal cells or on cells that are undesirable to be treated).

[0462] For example, in some embodiments, the cells comprise a receptor expressing a first engineered antigen receptor (e.g., CAR or TCR) that is capable of inducing an activating signal for the cell, typically upon specific binding to an antigen recognized by the first receptor, for example, with the first antigen. In some embodiments, the cell further comprises a second engineered antigen receptor (e.g., CAR or TCR), e.g., a chimeric co-stimulatory receptor, that is capable of inducing a co-stimulatory signal for an immune cell, typically upon specific binding to a second antigen recognized by the second receptor. . In some embodiments, the first antigen and the second antigen are the same. In some embodiments, the first antigen and the second antigen are different.

[0463] In some embodiments, the first and/or second engineered antigen receptor (eg, CAR or TCR) is capable of inducing an activating signal for the cell. In some embodiments, the receptor contains an intracellular signaling component containing ITAM motifs or ITAM-like motifs. In some embodiments, activation induced by the first receptor comprises signal transduction or a change in protein expression in a cell resulting in initiation of an immune response such as ITAM phosphorylation and/or initiation of an ITAM-mediated signal transduction cascade, formation of an immunological synapse, and/or formation of a cluster of molecules near the associated receptor (e.g., CD4 or CD8, and the like), activating one or more transcription factors such as NF-κB and/or AP-1, and/or inducing gene expression for factors such as cytokines, proliferation and/or survival.

[0464] In some embodiments, the first and/or second receptor comprises the intracellular signaling domains of co-stimulatory receptors such as CD28, CD137 (4-1 BB), OX40, and/or ICOS. In some embodiments, the first and second receptors contain intracellular co-stimulatory receptor signaling domains that are different. In one embodiment, the first receptor contains the CD28 costimulatory signaling region and the second receptor contains the 4-1BB costimulatory signaling region, or vice versa.

[0465] In some embodiments, the first and/or second receptor includes both an intracellular signaling domain containing ITAM or ITAM-like motifs and an intracellular signaling domain of a co-stimulatory receptor.

[0466] In some embodiments, the first receptor contains an intracellular signaling domain containing ITAM or ITAM-like motifs, and the second receptor contains an intracellular signaling domain of a costimulatory receptor. The costimulatory signal, in combination with an activating signal induced in the same cell, results in an immune response, such as a sustained and long-lasting immune response, such as an increase in gene expression, secretion of cytokines and other factors, and T-lymphocyte-mediated effector functions, such as cell destruction.

[0467] In some embodiments, neither ligation of the first receptor per se nor ligation of the second receptor per se induces a sustained immune response. In some aspects, if only one receptor is ligated, the cell is tolerized or becomes tolerant to the antigen or inhibited, and/or does not induce proliferation or secretion factors, or does not perform effector functions. In some such embodiments, however, when a plurality of receptors are ligated, for example upon encountering a cell expressing the first and second antigens, the desired response is achieved, such as complete immune activation or stimulation, for example, as indicated by the secretion of one or more cytokines, proliferation, persistence and/or implementation of an immune effector function such as cytotoxic killing of target cells.

[0468] In some embodiments, cells expressing the recombinant receptor further comprise inhibitory CARs (iCARs, see Fedorov et al., Sci. Transl. Medicine, 5(215) (2013), such as CARs that recognize an antigen other than an antigen associated with and/or specific to a disease or condition, wherein the activating signal from a non-disease-targeting CAR is reduced or inhibited by binding the inhibitory CAR to its ligand, for example, to reduce off-target effects.

[0469] In some embodiments, the two receptors induce, respectively, an activating and inhibitory signal to a cell, such that binding of one of the receptors to its antigen activates the cell or induces a response, but binding of the second inhibitory receptor to its antigen induces a signal that suppresses or dampens such a response. Examples are combinations of activating CARs and inhibitory CARs or iCARs. For example, a strategy can be used in which an activating CAR binds an antigen that is expressed in the disease or condition but is also expressed on normal cells, and an inhibitory receptor binds a single antigen that is expressed on normal cells but not on cells of the disease or condition.

[0470] In some aspects, the chimeric receptor is or contains an inhibitory CAR (eg, iCAR) and contains intracellular components that dampen or suppress an immune response, such as an ITAM and/or costimulatory-promoted response in a cell. Examples of such intracellular signaling components are those found on immune checkpoint molecules, including PD-1, CTLA4, LAG3, BTLA, OX2R, TIM-3, TIGIT, LAIR-1, PGE2 receptors, EP2/4 adenosine receptors, including A2AR . In some aspects, the engineered cell contains an inhibitory CAR comprising or derived from the signaling domain of such inhibitory molecule, such that it serves to dampen the response of the cell, for example, that induced by an activating and/or co-stimulatory CAR.

[0471] In some embodiments, a multitargeting strategy is used when an antigen associated with a particular disease or condition is expressed on a non-disease cell and/or is expressed on the genetically engineered cell itself, either transiently (e.g., when stimulated in connection with a gene engineering), or permanently. In such cases, if the ligation of two separate and individually specific antigen receptors is required, specificity, selectivity and/or efficacy can be improved.

[0472] In some embodiments, a plurality of antigens, such as first and second antigens, are expressed in a cell, tissue, or disease or condition that is targeted, such as on a cancer cell. In some aspects, the cell, tissue, disease, or condition is multiple myeloma or a multiple myeloma cell. In some embodiments, one or more of the plurality of antigens is typically also expressed on a cell that is not the desired target of cell therapy, such as a normal or non-diseased cell or tissue, and/or the genetically engineered cells themselves. In such embodiments, if multiple receptor ligation is required to achieve cell responses, specificity and/or efficacy is achieved.

B. Cells and preparation of cells for genetic engineering

[0473] Among the cells expressing the receptors and introduced using the proposed methods, there are genetically engineered cells. Genetic engineering typically involves introducing a nucleic acid encoding a recombinant or genetically engineered component into a composition containing cells, for example, by retroviral transduction, transfection, or transformation.

[0474] In some embodiments, the nucleic acids are heterologous, that is, not normally present in a cell or in a sample derived from a cell, such as a sample derived from another organism or cell, which, for example, is not normally found in the genetic engineered cell and/or in the organism from which such a cell is obtained. In some embodiments, the nucleic acids are not naturally occurring, for example, the nucleic acid is not naturally occurring, including nucleic acid containing chimeric combinations of nucleic acids encoding different domains from many different cell types.

[0475] The cells are typically eukaryotic cells, such as mammalian cells, and typically are human cells. In some embodiments, the cells are derived from blood, bone marrow, lymph, or lymphoid organs and are cells of the immune system, such as cells of innate or adaptive immunity, e.g., myeloid or lymphoid cells, including lymphocytes, typically T-lymphocytes and /or NK cells. Other exemplary cells include stem cells such as multipotent and pluripotent stem cells, including induced pluripotent stem cells (iPSCs). The cells are typically primary cells, such as cells isolated directly from the subject and/or isolated from the subject and frozen. In some embodiments, the cells contain one or more subsets of T lymphocytes or other cell types, such as whole populations of T lymphocytes, CD4+ cells, CD8+ cells, and subpopulations thereof, e.g., defined by function, activation state, maturity, potentials for differentiation , replication, recycling, localization, and/or persistence capabilities, type of antigen specificity, type of antigen receptor, presence in a particular organ or compartment, cytokine secretion marker or profile, and/or degree of differentiation. With respect to the subject to be treated, the cells may be allogeneic and/or autologous. The methods include standard methods. In some aspects, for example, for standard technologies, the cells are pluripotent and/or multipotent, for example, they are stem cells, such as induced pluripotent stem cells (iPSC). In some embodiments, the methods include isolating cells from a subject, preparing, processing, culturing and/or genetically engineering them, and reintroducing them to the same subject, before or after cryopreservation.

[0476] Among the subtypes and subpopulations of T lymphocytes and/or CD4+ and/or CD8+ T lymphocytes are naïve T lymphocytes (T _N ), effector T lymphocytes (T _EFF ), memory T lymphocytes and their subtypes, such as memory T-cell stem cells (T _SCM ), central memory T-lymphocytes (T _CM ), effector memory T-lymphocytes (T _EM ) or terminally differentiated effector memory T-lymphocytes, antitumor effector lymphocytes (TIL), immature T-lymphocytes, mature T lymphocytes, helper T lymphocytes, cytotoxic T lymphocytes, mucosal-associated invariant T lymphocytes (MAIT), naturally occurring and adaptive regulatory T lymphocytes (Treg), helper T lymphocytes such as TH1 cells, cells TH2, TH3 cells, TH17 cells, TH9 cells, TH22 cells, follicular helper T lymphocytes, alpha/beta T lymphocytes, and delta/gamma T lymphocytes.

[0477] In some embodiments, the cells are natural killer (NK) cells. In some embodiments, the cells are monocytes or granulocytes, such as myeloid cells, macrophages, neutrophils, dendritic cells, mast cells, eosinophils, and/or basophils.

[0478] In some embodiments, the cells contain one or more genetically engineered nucleic acids and express recombinant or genetically engineered products of such nucleic acids. In some embodiments, nucleic acids are heterologous, that is, not normally present in a cell or in a sample derived from a cell, such as derived from another organism, or cells that, for example, are not normally found in a genetically engineered cell and /or in the organism from which such a cell is obtained. In some embodiments, nucleic acids are not naturally occurring, such as non-naturally occurring nucleic acids, including nucleic acids containing chimeric combinations of nucleic acids encoding different domains from many different cell types.

[0479] In some embodiments, obtaining genetically engineered cells includes one or more stages of cultivation and/or preparation. Cells for the introduction of a nucleic acid encoding a transgenic receptor, such as CAR, can be isolated from a sample, such as a biological sample, for example, a sample obtained or isolated from a subject. In some embodiments, the subject from which cells are isolated is one who has a disease or condition, or is in need of cell therapy, or is being administered cell therapy. The subject, in some embodiments, is a human in need of a particular therapeutic intervention, such as an adoptive cell therapy for which cells are isolated, processed, and/or genetically engineered.

[0480] Accordingly, the cells, in some embodiments, are primary cells, eg, primary human cells. Samples include tissue, fluid and other samples taken directly from the subject, as well as samples obtained as a result of one or more stages. treatments such as separation, centrifugation, genetic engineering (eg transduction with a viral vector), washing and/or incubation. The biological sample may be a sample obtained directly from a biological source or a sample that is being processed. Biological samples include, but are not limited to, bodily fluids such as blood, plasma, serum, cerebrospinal fluid, synovial fluid, urine and sweat, tissue and organ samples, including processed samples derived therefrom.

[0481] In some aspects, the sample from which cells are obtained or isolated is blood or a sample obtained from blood, or is a product or product obtained from apheresis or leukapheresis. Illustrative specimens include whole blood, peripheral blood mononuclear cells (PBMCs), leukocytes, bone marrow, thymus, tissue biopsy, tumor, leukemia, lymphoma, lymph node, gastrointestinal-associated lymphoid tissue, mucosal-associated lymphoid tissue, spleen, other lymphoid tissues, liver, lung, stomach, intestines, colon, kidneys, pancreas, breast, bones, prostate, cervix, testes, ovaries, palatine tonsil or other organ, and/or cells derived from them. Samples include, in the context of cell therapy, such as adoptive cell therapy, samples from autologous and allogeneic sources.

[0482] In some embodiments, the cells are derived from cell lines, such as T-lymphocyte lines. The cells, in some embodiments, are derived from a xenogeneic source, such as mice, rats, non-human primates, and pigs.

[0483] In some embodiments, cell isolation includes one or more steps of preparing and/or separating cells not based on affinity. In some examples, cells are washed, centrifuged, and/or incubated in the presence of one or more reagents, for example, to remove unwanted components, enrich in desirable components, lyse, or remove cells sensitive to particular reagents. In some examples, cells are separated based on one or more properties such as density, adhesion properties, size, sensitivity, and/or resistance to particular components.

[0484] In some examples, cells from the subject's bloodstream are obtained, for example, by apheresis or leukapheresis. Samples, in some aspects, contain lymphocytes, including T-lymphocytes, monocytes, granulocytes, B-lymphocytes, other nucleated leukocytes, erythrocytes and/or platelets, and in some aspects contain cells other than erythrocytes and platelets.

[0485] In some embodiments, the blood cells collected from the subject are washed, for example, to remove a plasma fraction and to place the cells in an appropriate buffer or medium for subsequent steps. processing. In some embodiments, the cells are washed with phosphate buffered saline (PBS). In some embodiments, the wash solution is free of calcium and/or magnesium and/or many or all of the divalent cations. In some aspects, the washing step is carried out in a semi-automated "through-flow" centrifuge (eg, Cobe 2991 cell processor, Baxter) according to the manufacturer's instructions. In some aspects, the washing step is carried out using flow filtration along the flow (TFF) according to the manufacturer's instructions. In some embodiments, the cells are resuspended in a variety of biocompatible buffers after washing, such as, for example, Ca-free PBS⁺⁺/Mg⁺⁺. In certain embodiments, the components of the blood cell sample are removed and the cells directly resuspended in the culture medium.

[0486] In some embodiments, the methods include methods for separating cells based on density, such as preparing leukocytes from peripheral blood by lysing erythrocytes and centrifuging in a Percoll or Ficoll gradient.

[0487] In some embodiments, isolation methods include separating different cell types based on the expression or presence of one or more specific molecules in the cell, such as surface markers, such as surface proteins, intracellular markers, or nucleic acid. In some embodiments, any known separation method based on such markers can be used. In some embodiments, the partition is a partition based on affinity or immunoaffinity. For example, isolation in some aspects includes separating cells and cell populations based on cell expression or the level of expression of one or more markers, typically cell surface markers, for example, by incubation with an antibody or binding partner that specifically binds to markers such as this is typically followed by washing steps and separating cells bound to the antibody or binding partner from those cells not bound to the antibody or binding partner.

[0488] Such separation steps may be based on positive selection, in which cells bound to the reagents are retained for further use, and/or negative selection, in which cells not bound to the antibody or binding partner are retained. In some examples, both fractions are retained for later use. In some aspects, negative selection can be particularly useful when there is no antibody available that specifically identifies a cell type in a heterogeneous population such that separation is best done based on markers expressed by cells other than the desired population.

[0489] Separation should not result in 100% enrichment or removal of a particular population of cells or cells expressing a particular marker. For example, positive selection or enrichment for cells of a particular type that, for example, express a marker refers to an increase in the number or percentage of such cells, but should not result in a complete absence of cells not expressing the marker. Thus, negative selection, removal or depletion of cells of a particular type, such as those expressing a marker, refers to a reduction in the number or percentage of such cells, but should not result in the complete removal of all such cells.

[0490] In some examples, a plurality of separation steps are performed when a positive or negative selection fraction from one stage is subjected to another separation step, such as a subsequent positive or negative selection. In some examples, a single separation step can deplete cells expressing multiple markers simultaneously, such as by incubating the cells with multiple antibodies or binding partners, each specific for the marker targeted for negative selection. Thus, multiple cell types can be positively selected simultaneously by incubating cells with multiple antibodies or binding partners expressed on different cell types.

[0491] For example, in some aspects, specific subpopulations of T-lymphocytes, such as cells positive, or expressing high levels of one or more surface markers, for example, CD28 ⁺ , CD62L ⁺ , CCR7 ⁺ , CD27 ⁺ , CD127 ⁺ , CD4+, CD8+ , CD45RA ⁺ , and/or CD45RO ⁺ T lymphocytes are isolated using positive or negative selection techniques.

[0492] For example, CD3 T lymphocytes⁺, CD28⁺ can be positively selected using conjugated magnetic anti-CD3/anti-CD28 beads (e.g. DYNABEADS® M-450 CD3/CD28 T Cell Expander).

[0493] In some embodiments, the implementation, the selection is carried out by enriching a particular population of cells through positive selection or depletion of a particular population of cells through negative selection. In some embodiments, positive or negative selection is performed by incubating cells with one or more antibodies or other binding agent that specifically binds to one or more surface markers expressed (marker ⁺ ) at a relatively high level (marker ^high ) on cells, obtained by positive or negative selection, respectively.

[0494] In some embodiments, T lymphocytes are isolated from a PBMC sample by negative selection for markers expressed on cells other than T lymphocytes, such as B lymphocytes, monocytes, or other white blood cells such as CD14. In some aspects, a CD4+ or CD8+ selection step is used to separate CD4+ helper T lymphocytes from CD8+ cytotoxic T lymphocytes. Such CD4+ and CD8+ populations can be further sorted into subpopulations by positive or negative selection for markers expressed or expressed at a relatively higher level in one or more subpopulations of naive T lymphocytes, memory T lymphocytes and/or effector T lymphocytes.

[0495] In some embodiments, CD8+ cells are further enriched or depleted in naïve cells, central memory cells, effector memory cells, and/or central memory stem cells, e.g., by positive or negative selection based on surface antigens associated with the respective subpopulation. . In some embodiments, memory central T lymphocytes (T _CM ) are enriched to increase efficacy, e.g., to improve long-term survival, multiplication, and/or engraftment after administration, which in some aspects are particularly persistent in such subpopulations. See Terakura et al., Blood.1:72-82 (2012); Wang et al., J Immunother. 35(9):689-701 (2012). In some embodiments, combining TCM-rich CD8+ T lymphocytes and CD4+ T lymphocytes further enhances efficacy.

[0496] In embodiments, memory T lymphocytes are present in subsets of CD8+ peripheral blood lymphocytes in both CD62L ⁺ and CD62L-. PBMCs can be enriched or depleted in CD62L-CD8+ and/or CD62L ⁺ CD8+ fractions, for example using anti-CD8 and anti-CD62L antibodies.

[0497] In some embodiments, the enrichment of central memory T lymphocytes (T _CM ) is based on positive or high surface expression of CD45RO, CD62L, CCR7, CD28, CD3 and/or CD 127; in some aspects, it is based on negative selection of cells expressing or strongly expressing CD45RA and/or granzyme B. In some aspects, isolation of a CD8+ population enriched in T _CM cells is accomplished by depletion of cells expressing CD4, CD14, CD45RA and positive selection or enrichment in cells expressing CD62L. In one aspect, enrichment for central memory T lymphocytes ( _TCM ) is performed by starting with a negative fraction of cells selected based on CD4 expression, which are negatively selected based on CD14 and CD45RA expression, and positively selected based on CD62L. These types of selection in some aspects are carried out simultaneously, and in other aspects are carried out sequentially, in any order. In some aspects, the same selection step based on CD4 expression as used in preparing a population or subset of CD8+ cells is also used to generate a population or subset of CD4+ cells such that both positive and negative fractions from the CD4 based separation are retained and used in subsequent steps of the methods, optionally after one or more additional positive or negative selection steps.

[0498] In a specific example, a PBMC sample or a sample of other leukocytes is subjected to selection of CD4+ cells, where both negative and positive fractions are retained. The negative fraction is then subjected to negative selection based on CD14 and CD45RA or CD19 expression and positive selection based on marker characteristics of central memory T lymphocytes such as CD62L or CCR7, where positive and negative selection is done in any order.

[0499] CD4+ T helper lymphocytes are sorted into naive cells, central memory cells, and effector cells by identifying cell populations that have cell surface antigens. CD4+ lymphocytes can be obtained using standard methods. In some embodiments, the naïve CD4+ T lymphocytes are CD45RO-, CD45RA ⁺ , CD62L ⁺ , CD4+ T lymphocytes. In some embodiments, the CD4+ central memory cells are CD62L ⁺ and CD45RO ⁺ . In some embodiments, the CD4+ effector cells are CD62L- and CD45RO-.

[0500] In one example, to enrich CD4+ cells through negative selection, the monoclonal antibody cocktail typically contains antibodies to CD14, CD20, CD11b, CD16, HLA-DR, and CD8. In some embodiments, the antibody or binding partner is bound to a solid support or matrix, such as a magnetic bead or paramagnetic bead, to allow cells to be separated for positive and/or negative selection. For example, in some embodiments, cells and cell populations are separated or isolated using immunomagnetic (or magnetoaffinity) separation techniques (discussed in Methods in Molecular Medicine, vol. 58: Metastasis Research Protocols, Vol. 2: Cell Behavior In vitro and In vivo , p 17-25 Edited by: S. A. Brooks and U. Schumacher © Humana Press Inc., Totowa, NJ).

[0501] In some aspects, a sample or composition of cells to be separated is incubated with a finely divided magnetizable or magnetically sensitive material such as magnetically sensitive particles or microparticles such as paramagnetic beads (such as Dynalbeads or MACS beads). The magnetically sensitive material, e.g., particles, is typically attached directly or indirectly to a binding partner, e.g., an antibody, that specifically binds to a molecule, e.g. , they wish to subject them to negative or positive selection.

[0502] In some embodiments, the magnetic particle or bead comprises a magnetically sensitive material associated with a specific binding element, such as an antibody or other binding partner. There are many well known magnetically sensitive materials used in magnetic separation processes. Suitable magnetic particles include those described in Molday US Pat. No. 4,452,773 and EP 452342 B, which are hereby incorporated by reference. Particles of colloidal size, such as those described in Owen, US patent No. 4795698, and Liberti et al., US patent No. 5200084, are other examples.

[0503] Incubation is generally carried out under conditions under which antibodies or binding partners, or molecules such as secondary antibodies or other reagents that specifically bind to such antibodies or binding partners that are attached to the magnetic particle or bead, bind specifically to molecules on the cell surface if they are present on the cells in the sample.

[0504] In some aspects, the sample is placed in a magnetic field, and cells containing magnetically sensitive or magnetizable particles attached to them will be attracted to the magnet and separated from unlabeled cells. For positive selection, cells that are attracted to a magnet are retained; for negative selection, cells that are not attracted (unlabeled cells) are retained. In some aspects, the combination of positive and negative selection is performed during the same selection step, where the positive and negative fractions are retained and further processed or subjected to additional separation steps.

[0505] In certain embodiments, the magnetically sensitive particles are coated with primary antibodies or other binding partners, secondary antibodies, lectins, enzymes, or streptavidin. In certain embodiments, the magnetic particles are attached to cells via a coating of primary antibodies specific for one or more markers. In certain embodiments, cells, rather than beads, are labeled with a primary antibody or binding partner, and then magnetic particles coated with a cell type-specific secondary antibody or other binding partner (eg, streptavidin) are added. In certain embodiments, streptavidin-coated magnetic particles are used in combination with biotinylated primary or secondary antibodies.

[0506] In some embodiments, the magnetically sensitive particles are left attached to cells that are to be subsequently incubated, cultured, and/or genetically engineered; in some aspects, the particles remain attached to the cells for administration to the patient. In some embodiments, the magnetizable or magnetically sensitive particles are removed from the cells. Methods for removing magnetizable particles from cells are known and include, for example, the use of competing unlabeled antibodies and magnetizable particles or antibodies conjugated to cleavable linkers. In some embodiments, the magnetizable particles are biodegradable.

[0507] In some embodiments, affinity selection is performed using magnetically activated cell sorting (MACS) (Miltenyi Biotec, Auburn, CA). Magnetic Activated Cell Sorting (MACS) systems are capable of selecting high purity cells containing magnetized particles attached to them. In certain embodiments, the MACS operates in a mode where non-target and target particles are sequentially eluted following the application of an external magnetic field. That is, the cells attached to the magnetic particles are held in place while the unattached particles are eluted. Then, after this first elution step is completed, the particles that are trapped in the magnetic field and prevented from being eluted are released in some way so that they can be eluted. In certain embodiments, non-target cells are labeled and depleted in a heterogeneous population of cells.

[0508] In certain embodiments, the isolation or separation is performed using a system, device, or apparatus that performs one or more of the isolation, cell preparation, separation, processing, incubation, culture, and/or preparation method steps. In some aspects, the system is used to perform each of these steps in a closed or sterile environment, such as to minimize error, user mismanagement, and/or contamination. In one example, the system is as described in PCT Publication Number WO2009/072003 or US Patent Application No. 20110003380 A1.

[0509] In some embodiments, the system or device performs one or more, for example, all of the steps of isolation, processing, genetic engineering, and preparation in an integrated or self-contained system and/or in an automated or programmable manner. In some aspects, the system or device includes a computer and/or a computer program in communication with the system or device that allows the user to program, control, evaluate the result and/or adjust various aspects of the processing, isolation, genetic engineering and preparation steps.

[0510] In some aspects, separation and/or other steps are performed using the CliniMACS system (Miltenyi Biotec), for example, to automatically separate cells at the clinical scale level in a closed and sterile system. Components may include an embedded microcomputer, a magnetic separation unit, a peristaltic pump, and various shut-off valves. The built-in computer in some aspects controls all the components of the tool and directs the system to carry out repetitive procedures in a standardized sequence. The magnetic separation assembly in some aspects includes a movable permanent magnet and a selection column holder. The peristaltic pump controls the flow rate through the tubing set and, together with check valves, provides a controlled flow of buffer through the system and continuous cell suspension.

[0511] The CliniMACS system in some aspects uses antibody-bound magnetizable particles that are supplied in a sterile, non-pyrogenic solution. In some embodiments, after labeling cells with magnetic particles, the cells are washed to remove excess particles. The cell preparation bag is then connected to a tubing set, which in turn is connected to a bag containing buffer and a cell collection bag. The tubing set consists of pre-assembled sterile tubing, including pre-column and separation column, and is intended for single use. After starting the separation program, the system automatically introduces the cell sample into the separation column. Labeled cells are retained on the column while unlabeled cells are removed through a series of washing steps. In some embodiments, cell populations for use with the methods described herein are unlabeled and are not retained on the column. In some embodiments, cell populations for use in conjunction with the methods described herein are labeled and retained in a column. In some embodiments, cell populations for use with the methods described herein are eluted from the column after removal of the magnetic field and collected in a cell collection bag.

[0512] In certain embodiments, separation and/or other steps are performed using a CliniMACS Prodigy system (Miltenyi Biotec). The CliniMACS Prodigy system is in some aspects provided with a cell processing unit that enables automatic washing and fractionation of cells by centrifugation. The CliniMACS Prodigy system may also include an integrated camera and programmable image recognition device that determines the optimal cell fractionation end point by detecting differences in macroscopic layers of the original cell product. For example, peripheral blood is automatically separated into erythrocytes, leukocytes, and plasma layers. The CliniMACS Prodigy system may also include an integrated cell culture chamber that performs cell culture protocols such as cell differentiation and expansion, antigen loading, and long-term cell culture. Entry nodes can allow sterile removal and exchange of media, and cells can be observed using an on-board microscope. See, for example, Klebanoff et al., J Immunother. 35(9): 651-660 (2012), Terakura et al., Blood.1:72-82 (2012) and Wang et al., J Immunother. 35(9):689-701 (2012).

[0513] In some embodiments, the cell populations described herein are collected and enriched (or depleted) using flow cytometry, in which cells stained with a plurality of cell surface markers are carried in a fluid stream. In some embodiments, the cell population described herein is harvested and enriched (or depleted) by preparative scale (FACS) sorting. In certain embodiments, the cell population described herein is harvested and enriched (or depleted) using microelectromechanical systems (MEMS) chips in combination with a FACS-based detection system (see, for example, WO 2010/033140, Cho et al. , Lab Chip 10, 1567-1573 (2010) and Godin et al., J Biophoton 1(5):355-376 (2008) In both cases, cells can be labeled with a variety of markers to enable well-defined subsets of T - lymphocytes of high purity.

[0514] In some embodiments, antibodies or binding partners are labeled with one or more detectable markers to facilitate separation for positive and/or negative selection. For example, separation may be based on the binding of fluorescently labeled antibodies. In some examples, cell separation based on the binding of antibodies or other binding partners specific for one or more cell surface markers is carried out in a fluid stream, for example, using a fluorescence excited cell sorter (FACS), including (FACS) in preparative scale and/or microelectromechanical systems (MEMS) chips, for example in combination with a flow cytometric detection system. Such methods allow positive and negative selection based on multiple markers at the same time.

[0515] In some embodiments, the preparation methods include freezing steps, such as cryopreservation of cells, either before or after isolation, incubation, and/or genetic engineering. In some embodiments, the freezing and subsequent thawing step removes granulocytes and, to some extent, monocytes in the cell population. In some embodiments, the cells are suspended in a freezing solution, for example, after a washing step to remove plasma and platelets. Any of a variety of known freezing solutions and parameters may be used in some aspects. One example involves using PBS containing 20% DMSO and 8% human serum albumin (HSA) or other appropriate cell freezing media. It is then diluted with media 1:1 so that the final concentration of DMSO and HSA is 10% and 4%, respectively. The cells are then typically frozen to -80° C. at a rate of 1° per minute and stored in the vapor phase in a liquid nitrogen storage tank.

[0516] In some embodiments, cells are incubated and/or cultured prior to or in association with genetic engineering. Incubation steps may include seeding, culturing, stimulating, activating and/or growing. Incubation and/or genetic engineering may be carried out in a culture vessel such as a node, chamber, well, column, tube, tube set, valve, vial, culture dish, bag or other container for seeding or culturing cells. In some embodiments, the compositions or cells are incubated in the presence of stimulating conditions or a stimulating agent. Such conditions include conditions designed to induce, proliferate, expand, activate, and/or survive cells in a population, to mimic antigen exposure, and/or to prime cells for genetic engineering, for example, to introduce a recombinant antigen receptor.

[0517] Conditions may include one or more specific environments, temperature, oxygen content, carbon dioxide content, time, agents such as nutrients, amino acids, antibiotics, ions, and/or stimulatory factors such as cytokines, chemokines, antigens, partners binding, fusion proteins, recombinant soluble receptors, and any other agents designed to activate cells.

[0518] In some embodiments, the stimulating conditions or agents include one or more agents, such as a ligand, that can activate an intracellular signaling domain of the TCR complex. In some aspects, the agent turns on or initiates an intracellular TCR/CD3 signaling cascade in a T lymphocyte. Such agents may include antibodies, eg specific for TCR, eg anti-CD3. In some embodiments, the stimulation conditions include one or more agents, such as a ligand, that can stimulate a co-stimulatory receptor, such as anti-CD28. In some embodiments, such agents and/or ligands can bind to a solid carrier, such as a bead, and/or one or more cytokines. Optionally, the propagation method may further comprise the step of adding the anti-CD3 and/or anti-CD28 antibody to the culture medium (eg, at a concentration of at least about 0.5 ng/mL). In some embodiments, the stimulatory agents include IL-2, IL-15 and/or IL-7. In some aspects, the concentration of IL-2 is at least about 10 units/ml.

[0519] In some aspects, incubation is carried out according to techniques such as those described in US patent No. 6040177, Riddell et al., Klebanoff et al., J Immunother. 35(9): 651-660 (2012), Terakura et al., Blood.1:72-82 (2012) and/or Wang et al., J Immunother. 35(9):689-701 (2012).

[0520] In some embodiments, T lymphocytes are propagated by adding feeder cells, such as non-dividing peripheral blood mononuclear cells (PBMCs) to the initiating culture composition (e.g., such that the resulting cell population contains at least about 5 , 10, 20, or 40 or more PBMC feeder cells for each T-lymphocyte in the initial population to proliferate); and incubating the culture (eg, for a period of time sufficient for these numbers of T-lymphocytes to multiply). In some aspects, non-dividing feeder cells may contain gamma-irradiated PBMC feeder cells. In some embodiments, the PBMC is irradiated with gamma radiation in the range of about 3000-3600 rad to prevent cell division. In some aspects, the feeder cells are added to the culture medium prior to the addition of the T lymphocyte populations.

[0521] In some embodiments, the stimulation conditions include a temperature appropriate for human T-lymphocyte growth, e.g., at least about 25 degrees Celsius, typically at least about 30 degrees, and typically at or about 37 degrees Celsius. Optionally, the incubation may further include the addition of non-dividing EBV-transformed lymphoblastic cells (LCL) as feeder cells. LCLs can be irradiated with gamma radiation in the range of about 6000 to 10000 rads. LCL feeder cells in some aspects are provided in any appropriate amount, eg, at a ratio of LCL feeder cells to initial T lymphocytes of at least about 10:1.

[0522] In embodiments, antigen-specific T-lymphocytes, such as antigen-specific CD4+ and/or CD8+ T-lymphocytes, are obtained by stimulating naive or antigen-specific T-lymphocytes with antigen. For example, lines or clones of antigen-specific T lymphocytes can be generated for cytomegalovirus antigens by isolating T lymphocytes from infected subjects and stimulating the cells in vitro with the same antigen.

C. Nucleic acids, vectors and genetic engineering methods

[0523] In some embodiments, cells, such as T-lymphocytes, are genetically engineered to express the recombinant receptor. In some embodiments, genetic engineering is carried out by introducing nucleic acid molecules that encode a recombinant receptor. Also provided are nucleic acid molecules encoding a recombinant receptor and vectors or constructs containing such nucleic acids and/or nucleic acid molecules.

[0524] In some cases, the nucleic acid sequence encoding a recombinant receptor, for example, a chimeric antigen receptor (CAR), contains a signal sequence that encodes a signal peptide. In some aspects, the signal sequence may encode a signal peptide derived from a native polypeptide. In other aspects, the signal sequence may encode a heterologous or non-native signal peptide. In some embodiments, the signal peptide is derived from a transmembrane protein. In some examples, the signal peptide is derived from CD8a, CD33, or IgG. Non-limiting illustrative examples of signal peptides include, for example, the CD33 signal peptide shown in SEQ ID NO:21, the CD8a signal peptide shown in SEQ ID NO:75, or the signal peptide shown in SEQ ID NO:76, or a modified version thereof. .

[0525] In some embodiments, the nucleic acid molecule encoding the recombinant receptor contains at least one promoter that is operably linked to it to control expression of the recombinant receptor. In some examples, the nucleic acid molecule contains two, three or more promoters operably linked to it to control expression of the recombinant receptor. In some embodiments, the nucleic acid molecule may contain regulatory sequences, such as transcription and translation initiation codons and stop codons, that are specific to the type of host (e.g., bacteria, fungus, plant, or animal) into which the nucleic acid molecule is to be introduced, appropriately and considering whether the nucleic acid molecule is based on DNA or RNA. In some embodiments, the nucleic acid molecule may contain regulatory/control elements such as a promoter, an enhancer gene, an intron, a polyadenylation signal, a Kozak consensus sequence, and a splicing acceptor or donor. In some embodiments, the nucleic acid molecule may contain a non-native promoter operably linked to a nucleotide sequence encoding a recombinant receptor and/or one or more additional polypeptides. In some embodiments, the promoter is selected from the pol I, pol II, or pol III RNA promoter. In some embodiments, the promoter is recognized by RNA polymerase II (eg, CMV, SV40 early region, or adenovirus major late promoter). In another embodiment, the promoter is recognized by RNA polymerase III (eg, the U6 or H1 promoter). In some embodiments, the promoter may be a non-viral promoter or a viral promoter such as the cytomegalovirus (CMV) promoter, the SV40 promoter, the RSV promoter, and a promoter found in the long terminal repeat of a mouse stem cell virus. Other known promoters are also contemplated.

[0526] In some embodiments, the promoter is or contains a constitutive promoter. Exemplary constitutive promoters include, for example, the simian virus 40 (SV40) early promoter, the cytomegalovirus immediate-early (CMV) promoter, the human ubiquitin C (UBC) promoter, the human elongation factor 1α (EF1α) promoter, the mouse phosphoglycerate kinase 1 (PGK) promoter, and chick β-actin promoter linked to the early CMV enhancer gene (CAGG). In some embodiments, the constitutive promoter is a synthetic or modified promoter. In some embodiments, the promoter is or contains the MND promoter, a synthetic promoter that contains the LTR region of MoMuLV U3 modified with a myeloproliferative sarcoma virus enhancer gene (see Challita et al. (1995) J. Virol. 69(2) :748-755). In some embodiments, the promoter is a tissue-specific promoter. In another embodiment, the promoter is a viral promoter. In another embodiment, the promoter is a non-viral promoter.

[0527] In another embodiment, the promoter is a regulated promoter (eg, an inducible promoter). In some embodiments, the promoter is an inducible promoter or a repressible promoter. In some embodiments, the promoter comprises a Lac operator sequence, a tetracycline operator sequence, a galactose operator sequence, or a doxycycline operator sequence, or is an analog thereof, or is capable of binding to or being recognized by the Lac repressor or the tetracycline repressor or an analog thereof. In some embodiments, the nucleic acid molecule does not contain a regulatory element, such as a promoter.

[0528] In some embodiments, a nucleic acid molecule encoding a recombinant receptor, e.g., CAR or other antigen receptor, further comprises nucleic acid sequences encoding a marker and/or cells expressing CAR or other antigen receptor, further comprises a marker, e.g., a surrogate marker, such as a cell surface marker, which can be used to confirm transduction or genetic engineering of a cell to express a receptor, such as a truncated version of a cell surface receptor, such as truncated EGFR (tEGFR). In some embodiments, one or more of the markers is a transduction marker, a surrogate marker, and/or a selection marker.

[0529] In some embodiments, the marker is a transduction marker or a surrogate marker. A transduction marker or surrogate marker can be used to detect cells that are introduced with a nucleic acid molecule, for example, a nucleic acid molecule encoding a recombinant receptor. In some embodiments, the transduction marker may indicate or confirm cell modification. In some embodiments, the surrogate marker is a protein that is designed to be co-expressed on the cell surface with a recombinant receptor, such as CAR. In specific embodiments, such a surrogate marker is a surface protein that is modified to have little or no activity. In certain embodiments, the surrogate marker is encoded by the same nucleic acid molecule that encodes the recombinant receptor. In some embodiments, a nucleic acid sequence encoding a recombinant receptor is operably linked to a nucleic acid sequence encoding a marker, optionally separated by an internal ribosome entry site (IRES), or to a nucleic acid encoding a self-cleaving peptide or a peptide that causes ribosome skipping. , such as a 2A sequence such as T2A, P2A, E2A, or F2A. External marker genes can in some cases be used in connection with a genetically engineered cell to allow detection or selection of cells and, in some cases, also facilitate cell suicide.

[0530] Exemplary surrogate markers may comprise truncated forms of cell surface polypeptides, such as truncated forms that are non-functional and do not or cannot transduce a signal or signal normally transduced by the full-length form of the cell surface polypeptide and/or are not internalized or unable to be internalized . Exemplary truncated cell surface polypeptides include truncated forms of growth factors or other receptors such as truncated human epidermal growth factor receptor 2 (tHER2), truncated epidermal growth factor receptor (tEGFR, exemplary tEGFR sequence shown in SEQ ID NO:11 or 76), or prostate-specific membrane antigen (PSMA) or a modified form thereof. The tEGFR may contain an epitope recognized by the cetuximab antibody (Erbitux®) or other anti-EGFR therapeutic antibody or binding molecule that can be used to identify or select genetically engineered cells that are derived from the tEGFR construct and the encoded exogenous protein, and/or to removal or separation of cells expressing the encoded exogenous protein. See US Pat. No. 8,802,374 and Liu et al., Nature Biotech. April 2016; 34(4): 430-434). In some aspects, the marker, eg, a surrogate marker, comprises all or part (eg, a truncated form) of CD34, NGFR, CD19, or truncated CD19, eg, truncated non-human CD19, or an epidermal growth factor receptor (eg, tEGFR). In some embodiments, the marker is or contains a fluorescent protein such as green fluorescent protein (GFP), enhanced green fluorescent protein (EGFP) such as superfolded GFP (sfGFP), red fluorescent protein (RFP) such as tdTomato, mCherry , mStrawberry, AsRed2, DsRed or DsRed2, blue fluorescent protein (CFP), blue-green fluorescent protein (BFP), enhanced blue fluorescent protein (EBFP), and yellow fluorescent protein (YFP), and variants thereof, including species variants, monomeric variants and codon-optimized and/or enhanced variants of fluorescent proteins. In some embodiments, the marker is or contains an enzyme such as luciferase, the lacZ gene from E. coli, alkaline phosphatase, secreted embryonic alkaline phosphatase (SEAP), chloramphenicol acetyltransferase (CAT). Exemplary light emitting reporter genes include luciferase (luc), β-galactosidase, chloramphenicol acetyltransferase (CAT), β-glucuronidase (GUS), or variants thereof.

[0531] In some embodiments, the marker is a selection marker. In some embodiments, the selection marker is or contains a polypeptide that confers resistance to exogenous agents or drugs. In some embodiments, the selection marker is an antibiotic resistance gene. In some embodiments, the selection marker is an antibiotic resistance gene that confers antibiotic resistance in mammalian cells. In some embodiments, the selection marker is or comprises a puromycin resistance gene, a hygromycin resistance gene, a blasticidin resistance gene, a neomycin resistance gene, a geneticin resistance gene, or a zeocin resistance gene, or a modified form thereof.

[0532] In some aspects, the marker, eg, a surrogate marker, includes all or part (eg, a truncated form) of CD34, NGFR, or an epidermal growth factor receptor (eg, tEGFR). In some embodiments, a nucleic acid encoding a marker is operably linked to a polynucleotide encoding a linker sequence, such as a cleavable linker sequence, eg T2A. For example, the marker, and optionally the linker sequence, may be any group as described in PCT Publication No. WO2014031687. For example, the marker may be a truncated EGFR (tEGFR) that is optionally linked to a linker sequence, such as a cleavable T2A linker sequence. An exemplary polypeptide for truncated EGFR (e.g., tEGFR) contains the amino acid sequence shown in SEQ ID NO: 7 or 28, or an amino acid sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with SEQ ID NO: 7 or 28. An exemplary T2A linker sequence contains an amino acid sequence, shown in SEQ ID NO: 6, or an amino acid sequence that shows at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95 %, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 6.

[0533] In some embodiments, nucleic acid molecules encoding such CAR constructs further comprise a sequence encoded by T2A, a ribosomal hopping element, and/or a tEGFR sequence, for example, downstream of the sequence encoding the CAR. In some embodiments, the sequence encodes a T2A ribosome skipping element set forth in SEQ ID NO: 6, or an amino acid sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with SEQ ID NO: 6. In some embodiments, T lymphocytes expressing an antigen receptor (e.g. . for the detection of such cells (see, for example, US patent No. 8802374). In some embodiments, the sequence encodes a tEGFR sequence shown in SEQ ID NO: 7 or 28, or an amino acid sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with SEQ ID NO: 7 or 28.

[0534] In some embodiments, a single promoter may direct the expression of an RNA that contains a single open reading frame (ORF), two or three genes (e.g., encoding a molecule involved in metabolic pathway modulation and encoding a recombinant receptor) separated from each other sequences encoding a self-cleaving peptide (eg 2A sequences) or a protease recognition site (eg furin). Thus, the ORF encodes a single polypeptide, which, either during (in the case of 2A) or after translation, is processed into separate proteins. In some cases, a peptide such as T2A can cause a ribosome to skip (ribosomal skipping) the synthesis of a peptide bond at the C-terminus of element 2A, resulting in separation of the end of the 2A sequence and the next peptide downstream of it (see, e.g., de Felipe. Genetic Vaccines and Ther 2:13 (2004) and deFelipe et al Traffic 5:616-626 (2004)). Many 2A elements are known in the art. Examples of 2A sequences that can be used in the methods and nucleic acids described herein include, but are not limited to, 2A sequences of stomatitis virus (F2A, e.g., SEQ ID NO: 27), equine rhinitis virus A (E2A, e.g., SEQ ID NO: 26), Thosea asigna virus (T2A, e.g., SEQ ID NO: 6 or 23), and porcine teshovirus-1 (P2A, e.g., SEQ ID NO: 24 or 25) as described in US Patent Publication No. 20070116690.

[0535] In some embodiments, the marker is a molecule, such as a cell surface protein, not naturally occurring on T lymphocytes or not naturally occurring on the surface of T lymphocytes, or a portion thereof. In some embodiments, the molecule is a foreign molecule, such as a foreign protein, that is, a protein that is not recognized as "self" by the immune system of the host to which the cells will be adoptively transferred.

[0536] In some embodiments, the marker does not serve a therapeutic function and/or does not produce an effect other than use as a genetically engineered marker, for example, to select cells successfully obtained by genetic engineering. In other embodiments, the marker may be a therapeutic molecule or a molecule otherwise having some desired effect, such as a ligand for a cell to be encountered in vivo, such as a co-stimulatory molecule or an immune checkpoint molecule to enhance and/or dampen responses. cells during adoptive transfer and encounter with the ligand.

[0537] The introduction of nucleic acid molecules encoding a recombinant receptor into a cell can be carried out using any number of known vectors. Such vectors include viral and non-viral systems, including lentiviral and gamma-retrovirus systems, as well as transposon based systems such as PiggyBac or Sleeping Beauty based gene transfer systems. Illustrative methods include methods for transferring receptor-encoding nucleic acids, including viral, eg, retroviral or lentiviral, transduction, transposons, and electroporation.

[0538] In some embodiments, gene transfer is accomplished by first stimulating the cell, e.g., by combining it with a stimulus that induces a response, such as proliferation, survival, and/or activation, e.g., as measured by expression of a cytokine or activation marker, with followed by transduction of activating cells and expansion in culture to quantities sufficient for clinical applications.

[0539] In some embodiments, prior to or during gene transfer, cells are incubated or cultured in the presence of an immunomodulatory compound, such as lenalidomide, including any compound as described herein. In some embodiments, an immunomodulatory compound, such as lenalidomide, is added during a cell production process, such as during a CAR T-lymphocyte genetic engineering process. In some aspects, the presence of an immunomodulatory compound may improve the quality of the resulting cell population. In some aspects, an immunomodulatory compound, such as lenalidomide, may increase cell proliferation or proliferation or may alter one or more signaling pathways resulting in cells with a less differentiated or less activated surface phenotype despite demonstrating sufficient proliferation and/or effector function. .

[0540] In some contexts, overexpression of a stimulatory factor (eg, lymphokine or cytokine) can be toxic to the subject. Thus, in some contexts, genetically engineered cells contain gene segments that render the cells susceptible to negative selection in vivo, such as when administered by adoptive immunotherapy. For example, in some aspects, the cells are genetically engineered so that they can be eliminated by altering the condition of the patient to whom they are administered in vivo. A negatively selectable phenotype may result from the insertion of a gene that confers sensitivity to an administered agent, such as a compound. Negatively selectable genes include the herpes simplex virus type I thymidine kinase gene (HSV-I TK) (Wigler et al., Cell 2:223, 1977), which confers sensitivity to ganciclovir; cellular hypoxanthine phosphoribosyl transferase (HPRT) gene, cellular adenine phosphoribosyl transferase (APRT) gene, bacterial cytosine deaminase gene, (Mullen et al., Proc. Natl. Acad. Sci. USA. 89:33 (1992)).

[0541] In some embodiments, recombinant nucleic acids are transferred into cells using particles of recombinant infectious viruses, such as, for example, vectors derived from simian virus 40 (SV40), adenoviruses, adeno-associated virus (AAV). In some embodiments, recombinant nucleic acids are transferred to T lymphocytes using recombinant lentiviral vectors or retroviral vectors such as retroviral gamma vectors (see, for example, Koste et al. (2014) Gene Therapy 2014 Apr 3. doi: 10.1038/ gt.2014.25; Carlens et al. (2000) Exp Hematol 28(10): 1137-46; Alonso-Camibo et al. (2013) Mol Ther Nucl Acids 2, e93; Park et al., Trends Biotechnol. 2011 November 29 (11): 550-557.

[0542] In some embodiments, the retroviral vector has a long terminal repeat (LTR) sequence, e.g., a retroviral vector derived from Moloney murine leukemia virus (MoMLV), myeloproliferative sarcoma virus (MPSV), mouse embryonic stem cell virus (MESV), mouse stem cell virus (MSCV), spleen necrosis virus (SFFV), or adeno-associated virus (AAV). Most retroviral vectors are derived from murine retroviruses. In some embodiments, retroviruses include viruses derived from any source of avian or mammalian cells. Retroviruses are generally amphotropic, meaning that they can infect host cells of several species, including humans. In one embodiment, the gene to be expressed replaces the gag, pol and/or env retroviral sequences. A number of exemplary retroviral systems have been described (e.g., US Pat. Nos. 5,219,740; 6,207,453; 5,219,740; Miller and Rosman (1989) BioTechniques 7:980-990; Miller, A. D. (1990) Human Gene Therapy 1:5-14; Scarpa et al. (1991) Virology 180:849-852; Burns et al. (1993) Proc. Natl. Acad. Sci. USA 90:8033-8037; and Boris-Lawrie and Temin (1993) Cur. Opin. Genet. Develop. 3 :102-109.

[0543] Methods for lentiviral transduction are known. Illustrative methods are described, for example, in Wang et al. (2012) J. Immunother. 35(9): 689-701; Cooper et al. (2003) Blood. 101:1637-1644; Verhoeyen et al. (2009) Methods Mol Biol. 506:97-114; and Cavalieri et al. (2003) Blood. 102(2): 497-505.

[0544] In some embodiments, recombinant nucleic acids are transferred to T lymphocytes via electroporation (see, e.g., Chicaybam et al, (2013) PLoS ONE 8(3): e60298 and Van Tedeloo et al. (2000) Gene Therapy 7 (16): 1431-1437). In some embodiments, the recombinant nucleic acids are transferred to T lymphocytes via transposition (see, e.g., Manuri et al. (2010) Hum Gene Ther 21(4): 427-437; Sharma et al. (2013) Molec Ther Nucl Acids 2, e74 and Huang et al (2009) Methods Mol Biol 506: 115-126). Other methods for introducing and expressing genetic material in immune cells include calcium phosphate transfection (eg, as described in Current Protocols in Molecular Biology, John Wiley & Sons, New York. N.Y.), protoplast fusion, cationic liposomal mediated transfection; microparticle bombardment with tungsten particles (Johnston, Nature, 346: 776-777 (1990)); and co-precipitation of strontium phosphate and DNA (Brash et al., Mol. Cell Biol., 7: 2031-2034 (1987)).

[0545] Other approaches and vectors for transferring nucleic acids encoding recombinant products are described, for example, in Published International Patent Application No: WO2014055668 and US Patent No. 7446190.

[0546] In some embodiments, cells, such as T-lymphocytes, can be transfected either during or after expansion, such as with a T-lymphocyte receptor (TCR) or a chimeric antigen receptor (CAR). This transfection to introduce the gene of the desired receptor can be done with, for example, any suitable retroviral vector. The population of genetically modified cells can then be released from the initial stimulus (eg, a CD3/CD2 stimulus) and then stimulated with a second type of stimulus, eg, with a de novo administered receptor). This second type of stimulus may comprise an antigenic stimulus in the form of a peptide/MHC molecule, a recognized (cross-linking) ligand of a genetically introduced receptor (e.g., a natural CAR ligand), or any ligand (such as an antibody) that directly binds in frame to the new receptor (e.g. , recognizing constant regions in the receptor). See, for example, Cheadle et al, “Chimeric antigen receptors for T-cell based therapy” Methods Mol Biol. 2012; 907:645-66, or Barrett et al., Chimeric Antigen Receptor Therapy for Cancer Annual Review of Medicine Vol. 65:333-347 (2014).

[0547] In some cases, you can use a vector that does not require cells, such as T-lymphocytes, to be activated. In some of these cases, cells may be selected and/or transduced prior to activation. Thus, cells can be obtained by genetic engineering before or after cell culture, and in some cases, at the same time as culture, or during at least part of it.

[0548] In some aspects, the cells are subjected to additional genetic engineering to facilitate the expression of cytokines or other factors. Among the additional nucleic acids are, for example, genes for introduction, which are those genes that improve the effectiveness of therapy, for example, contributing to the viability and/or functioning of the transferred cells; genes for obtaining a genetic marker for selection and/or evaluation of cells, for example, for evaluation of in vivo survival or localization; genes to improve safety, for example making cells susceptible to negative selection in vivo, as described by Lupton S. D. et al., Mol. and Cell Biol., 11:6 (1991); and Riddell et al., Human Gene Therapy 3:319-338 (1992); see also PCT/US91/08442 and PCT/US94/05601, Lupton et al., describing the use of bifunctional selectable fusion genes derived from the fusion of a dominant positive selectable marker with a negative selectable marker. See, for example, Riddell et al., US Pat. No. 6,040,177, columns 14-17.

III. Illustrative Treatment Outcomes and How to Evaluate Them

[0549] In some embodiments of the methods, compositions, combinations, kits, and uses provided herein, the proposed combination therapy results in one or more treatment outcomes, such as a feature associated with any one or more therapy-related parameters, or treatment as described below. In some embodiments, the method includes assessing the exposure, persistence, and proliferation of T lymphocytes, eg, T lymphocytes administered for T lymphocyte therapy. In some embodiments, exposure or prolonged expansion and/or persistence of cells, and/or changes in cell phenotypes or functional activity of cells, e.g., cells administered for immunotherapy, e.g., T cell therapy, in the methods provided herein, can be measure by characterizing T-lymphocytes in vitro or ex vivo. In some embodiments, such assays can be used to determine or confirm T-lymphocyte function, such as T-cell therapy, before or after administration of the combination therapy provided herein.

[0550] In some embodiments, the combination therapy may further include one or more screening steps to identify subjects for treatment with the combination therapy and/or continuation of the combination therapy, and/or the step of evaluating treatment outcomes and/or monitoring treatment outcomes. In some embodiments, the stage of evaluating treatment outcomes may include the stages of evaluating and/or monitoring treatment and/or identifying subjects for the introduction of additional or remaining stages of therapy and/or for retreatment. In some embodiments, the screening and/or outcome evaluation step may be used to determine the dose, frequency, duration, timing, and/or sequencing of the combination therapy provided herein.

[0551] In some embodiments, any of the steps of screening and/or evaluating treatment outcomes described herein may be used before, during, during, or after the administration of one or more of the steps of the proposed combination therapy, e.g., administration of T- cell therapy (eg, CAR-expressing T-lymphocytes), and/or immunomodulatory compounds, such as lenalidomide. In some embodiments, the evaluation is done before, during, during the course, or after any of the methods provided herein. In some embodiments, the evaluation is done prior to the implementation of the methods provided herein. In some embodiments, the evaluation is done after one or more of the steps of the methods provided herein. In some embodiments, the evaluation is carried out prior to the introduction of one or more stages of the proposed combination therapy, for example, to screen and identify patients prone and/or susceptible to receive combination therapy. In some embodiments, evaluation is performed during, during, or after the administration of one or more steps of the proposed combination therapy, e.g., to evaluate intermediate or end points of treatment, e.g., to determine the effectiveness of treatment and/or to determine whether to continue or repeat treatment and/or to determine whether to administer the remaining stages of concomitant therapy.

[0552] In some embodiments, the results of treatment include an improvement in immune function, for example, the immune function of T lymphocytes administered for cell-based therapy and/or endogenous T lymphocytes in the body. In some embodiments, exemplary treatment outcomes include, but are not limited to, increased T-lymphocyte proliferation, increased functional activity of T-lymphocytes, changes in the expression of an immune cell phenotypic marker, for example, such features are associated with engineered T-lymphocytes, for example, CAR T-lymphocytes administered to a subject. In some embodiments, exemplary treatment outcomes include reduced disease severity, such as tumor mass, improved clinical outcomes, and/or increased efficacy of therapy.

[0553] In some embodiments, the step of screening and/or evaluating treatment outcomes includes evaluating the survival and/or function of T lymphocytes administered for cell-based therapy. In some embodiments, the step of screening and/or evaluating treatment outcomes includes evaluating levels of cytokines or growth factors. In some embodiments, the step of screening and/or evaluating treatment outcomes includes assessing disease severity and/or improvement, such as assessing tumor mass and/or clinical outcomes. In some embodiments, the step of either screening and/or evaluating treatment outcomes may include any of the evaluation methods and/or assays described herein and/or known in the art, and may be performed one or more times, for example, before, during course, during the course or after the introduction of one or more stages of combination therapy. Exemplary sets of treatment outcome-related parameters that may be assessed in some embodiments of the methods provided herein include a peripheral blood immune cell population profile and/or tumor mass.

[0554] In some embodiments, the methods affect the effectiveness of cell therapy for a subject. In some embodiments, the persistence, expansion, and/or presence of recombinant receptor-expressing cells, e.g., CAR-expressing cells, in a subject after dosing cells in a method with an immunomodulatory compound is greater than what is achieved with a method without administration. immunomodulatory compound. In some embodiments of the immunotherapy methods provided herein, such as T-cell therapy (e.g., CAR-expressing T-lymphocytes), the evaluation of parameters includes an assessment of proliferation and/or persistence in a subject receiving T-lymphocytes for immunotherapy, e.g. , T cell therapy, as compared to a method in which immunotherapy is administered to a subject without an immunomodulatory compound. In some embodiments, the methods result in administered T lymphocytes exhibiting increased or prolonged proliferation and/or persistence in the subject compared to a method in which the T cell therapy is administered to the subject without an immunomodulatory compound.

[0555] In some embodiments, administration of an immunomodulatory compound, such as lenalidomide, reduces disease severity, such as tumor mass, in a subject compared to a method in which a dose of cells expressing the recombinant receptor is administered to the subject without the immunomodulatory compound. In some embodiments, administration of an immunomodulatory compound, such as lenalidomide, reduces the number of bone marrow blast cells in a subject compared to a method in which a dose of cells expressing the recombinant receptor is administered to the subject without the immunomodulatory compound. In some embodiments, administration of an immunomodulatory compound, e.g., lenalidomide, results in improved clinical outcomes, e.g., objective response rate (ORR), progression-free survival (PFS), and overall survival (OS), compared to the method when wherein a dose of cells expressing the recombinant receptor is administered to a subject without an immunomodulatory compound.

[0556] In some embodiments, the subject may be screened prior to the introduction of one or more stages of combination therapy. For example, a subject may be screened for disease characteristics and/or disease severity, eg, tumor mass, prior to administration of combination therapy to determine compliance, responsiveness, and/or responsiveness to administration of combination therapy. In some embodiments, the screening and/or outcome evaluation step may be used to determine the dose, frequency, duration, timing, and/or sequencing of the combination therapy provided herein.

[0557] In some embodiments, the subject may be screened after the introduction of one of the stages of combination therapy, to determine and identify subjects for receiving the remaining stages of combination therapy and/or to monitor the effectiveness of therapy. In some embodiments, the number, level, or amount of T lymphocytes administered and/or the proliferation and/or activity of T lymphocytes administered is assessed before and/or after administration of an immunomodulatory compound, eg, lenalidomide.

[0558] In some embodiments, an immunomodulatory compound, such as lenalidomide, is administered until the concentration or number of genetically engineered cells in the subject's blood is (i) at least, or about at least 10 genetically engineered cells. engineered cells per microliter, (ii) at least 20%, 30%, 40%, or 50% of total peripheral blood mononuclear cells (PBMC), (iii) at least, or about at least 1 x 10 ⁵ engineered cells or (iv) at least 5,000 copies of DNA encoding the recombinant receptor per microgram of DNA; and/or on day 90 after the start of administration (a) CAR-expressing cells are detected in the subject's blood or serum; and/or on day 90 after the start of administration (a) the subject's blood contains at least 20% CAR-expressing cells, at least 10 CAR-expressing cells per microliter, or at least 1 x 10 ⁴ CAR- expressing cells.

[0559] In some embodiments, an immunomodulatory compound, e.g., lenalidomide, is administered until there is a clinical benefit from the treatment, such as at least a 50% or greater reduction in total tumor volume or a complete response (CR), at which the detectable tumor disappears, progression-free survival, or disease-free survival for more than 6 months or more than 1 year or more.

[0560] In some embodiments, a change and/or alternation is determined or evaluated, such as an increase, increase, decrease, or decrease in levels, values, or measurements of a parameter or result compared to levels, values, or measurements of the same parameter or result at another time. evaluation time, in another state, at the reference point and/or at another subject. For example, in some embodiments, a fold change, such as an increase or decrease in specific parameters, such as the number of engineered T lymphocytes in a sample, compared to the same parameter in another condition, such as before or after administration of an immunomodulatory compound, may be determined. eg lenalidomide. In some embodiments, the levels, values, or measurements of two or more parameters are determined and the relative levels are compared. In some embodiments, the determined levels, values, or measurements of parameters are compared to levels, values, or measurements from a control sample or an untreated sample. In some embodiments, certain levels, values, or parameter measurements are compared to levels from a sample from the same subject, but at a different point in time. Values derived from the quantification of an individual parameter may be combined for disease assessment purposes, for example, by forming an arithmetic or logical operation on the levels, values, or measurements of the parameters using multiparametric analysis. In some embodiments, a ratio of two or more specific parameters may be calculated.

A. Exposure, persistence and proliferation of T-lymphocytes

[0561] In some embodiments, a parameter associated with therapy or treatment outcome, which includes parameters that can be assessed for screening steps and/or for evaluating treatment outcomes and/or monitoring treatment outcomes, is or includes an assessment of exposure, persistence, and proliferation of T-lymphocytes, for example, T-lymphocytes administered for therapy based on T-lymphocytes. In some embodiments, increased exposure or prolonged proliferation and/or persistence of cells, and/or changes in cell phenotypes or functional activity of cells, e.g., cells administered for immunotherapy, e.g., T cell therapy, in the methods provided herein , can be measured by characterizing T-lymphocytes in vitro or ex vivo. In some embodiments, such assays can be used to determine or confirm the function of T-lymphocytes used for immunotherapy, for example, T-cell therapy, before or after the introduction of one or more stages of combination therapy proposed herein.

[0562] In some embodiments, administration of an immunomodulatory compound, e.g., lenalidomide, is designed to facilitate exposure of the subject to cells, e.g., T-lymphocytes, administered for T-lymphocyte therapy, e.g., by facilitating their expansion and/or persistence with time. In some embodiments, the T cell therapy demonstrates an increase or prolongation of proliferation and/or persistence in a subject compared to a method in which the T cell therapy is administered to the subject without an immunomodulatory compound, such as lenalidomide.

[0563] In some embodiments, the methods provide increased exposure of a subject to administered cells (eg, increased cell number or duration) and/or improved efficacy and therapeutic outcomes of immunotherapy, such as T cell therapy. In some aspects, the methods are advantageous in that higher and/or longer exposure for cells expressing recombinant receptors, such as CAR expressing cells, improves treatment outcomes over other methods. Such outcomes may include patient survival and remission, even in individuals with large tumor masses.

[0564] In some embodiments, administration of an immunomodulatory compound, e.g., lenalidomide, can increase the maximum exposure, total exposure, and/or duration of exposure for cells, e.g., T-lymphocytes administered for T-lymphocyte-based therapy, in a subject compared with the introduction of only T-lymphocytes without an immunomodulatory compound. In some aspects, administration of an immunomodulatory compound, such as lenalidomide, in the context of greater disease severity (and thus large amounts of antigen) and/or more aggressive or resistant cancer enhances efficacy compared to administration of T-lymphocytes alone without the immunomodulatory compound, in that same context, which may result in immunosuppression, weakness and/or depletion, which may prevent cell proliferation and/or persistence.

[0565] In some embodiments, the presence and/or amount of recombinant receptor-expressing cells (e.g., CAR-expressing cells administered for T-lymphocyte therapy) is detected in a subject after administration of T-lymphocytes and before, during, and /or after administration of an immunomodulatory compound, for example lenalidomide. In some aspects, quantitative PCR (qPCR) is used to evaluate the number of cells expressing a recombinant receptor (e.g., CAR-expressing cells administered for T-lymphocyte therapy) in a blood or serum sample, or in an organ or tissue sample (e.g., at the disease site, e.g., in a tumor sample) of the subject. In some aspects, persistence is quantified as copies of DNA or plasmid encoding a receptor, e.g., CAR, per microgram of DNA, or as the number of receptor-expressing, e.g., CAR-expressing cells per microliter of sample, e.g., blood or serum, or per total the number of peripheral blood mononuclear cells (PBMC) or leukocytes or T-lymphocytes per microliter of sample.

[0566] In some embodiments, cells are detected in a subject on or at least on 4, 14, 15, 27 or 28 days after the administration of T-lymphocytes, for example, CAR-expressing T-lymphocytes. In some aspects, cells are detected at or at least at 2, 4, or 6 weeks, or at 3, 6, or 12, 18, or 24, or 30, or 36 months, or at 1, 2, 3, 4, 5 or later one year after the administration of T-lymphocytes, for example, CAR-expressing T-lymphocytes, and/or an immunomodulatory compound, for example, lenalidomide.

[0567] In some embodiments, the persistence of receptor-expressing cells (e.g., CAR-expressing cells) in a subject according to the methods following administration of T lymphocytes, e.g., CAR-expressing T lymphocytes and/or an immunomodulatory compound, e.g., lenalidomide, more than what would be achieved with alternative methods, such as methods involving the administration of immunotherapy alone, for example, the administration of T-lymphocytes, for example, CAR-expressing T-lymphocytes, without an immunomodulatory compound.

[0568] Exposure, for example, the number of cells, for example, T-lymphocytes, administered for T-cell therapy, as an indicator of expansion and / or persistence can be formulated in terms of the maximum number of cells for which the subject is exposed, the duration of the presence of detectable cells, or cells above a certain amount or percentage, the area under the curve of the number of cells depending on time and / or their combinations and their indicators. Such results can be assessed using known methods such as qPCR to detect the copy number of a nucleic acid encoding a recombinant receptor compared to the total amount of nucleic acid or DNA in a particular sample, e.g. blood, serum, plasma or tissue, e.g. tumor sample, and/or using flow cytometric assays that detect cells expressing the receptor, usually using antibodies specific for these receptors. Cell-based assays can also be used to detect the number or percentage of functional cells, such as cells capable of binding and/or neutralizing and/or eliciting reactions, such as cytotoxic reactions, against cells of a disease or condition, or expressing an antigen recognized by a receptor.

[0569] In some aspects, increasing a subject's exposure to cells includes increasing cell proliferation. In some embodiments, receptor-expressing cells, eg, CAR-expressing cells, proliferate in the subject after administration of T lymphocytes, eg, CAR-expressing T lymphocytes, and/or after administration of an immunomodulatory compound, eg, lenalidomide. In some aspects, the method results in an increase in cell proliferation compared to other methods, for example, involving the administration of T lymphocytes, for example, CAR expressing T lymphocytes, without the administration of an immunomodulatory compound, for example, lenalidomide.

[0570] In some aspects, the method results in high in vivo proliferation of the administered cells, for example, as measured by flow cytometry. In some aspects, high peak cell proportions are detected. For example, in some embodiments, at a peak peak level following administration of T lymphocytes, e.g., CAR-expressing T lymphocytes, and/or an immunomodulatory compound, e.g. , in the PBMC fraction or in the T-lymphocyte fraction, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50% , at least about 60%, at least about 70%, at least about 80%, or at least about 90% of the cells express the recombinant receptor, eg, CAR.

[0571] In some embodiments, the method results in a maximum concentration, in the blood or serum or other bodily fluid or organ or tissue of the subject, of at least 100, 500, 1000, 1500, 2000, 5000, 10,000, or 15,000 copies, and of the nucleic acid itself encoding a receptor, for example, CAR, per microgram of DNA or at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0, 8 or 0.9 receptor-expressing, eg, CAR-expressing cells per total peripheral blood mononuclear cells (PBMC), total mononuclear cells, total T-lymphocytes, or total microliters. In some embodiments, cells expressing the receptor are detected at least 10%, 20%, 30%, 40%, 50%, or 60% of the total amount of PBMC in the subject's blood, and/or at the same level for at least , 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, 48, or 52 weeks after administration of T-lymphocytes, e.g., CAR-expressing T-lymphocytes, and/ or an immunomodulatory compound, for example, lenalidomide, or for 1, 2, 3, 4 or 5 or more years after such administration.

[0572] In some aspects, the method results in at least 2-fold, at least 4-fold, at least 10-fold, or at least 20-fold increase in copies of the nucleic acid encoding a recombinant receptor, eg, CAR, per microgram of DNA, eg, in serum, plasma, blood, or tissue, eg, in a tumor sample from a subject.

[0573] In some embodiments, cells expressing the receptor are detected in serum, plasma, blood, or tissue, e.g., in a tumor sample from a subject, e.g., using a specified method, such as qPCR or a flow cytometric detection method, by at least 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 days or more after administration of T-lymphocytes, such as CAR-expressing T -lymphocytes, or after the administration of an immunomodulatory compound, for example, lenalidomide, for at least, or about, at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 , 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 or more weeks after administration of T-lymphocytes, e.g., CAR-expressing T-lymphocytes, and/or immunomodulatory compound, e.g., lenalidomide .

[0574] In some aspects, at least about 1 x 10 ² , at least about 1 x 10 ³ , at least about 1 x 10 ⁴ , at least about 1 x 10 ⁵ , or, by at least about 1 x 10 ⁶ or at least about 5 x 10 ⁶ or at least about 1 x 10 ⁷ or at least about 5 x 10 ⁷ or at least about 1 x 10 ⁸ recombinant receptor expressing cells, e.g. CAR expressing cells, and/or at least 10, 25, 50, 100, 200, 300, 400 or 500 or 1000 receptor expressing cells per microliter, for example, at least 10 per microliter are detected or present in the subject or in its fluid, plasma, serum, tissue or compartment, for example, in the blood, for example, peripheral blood, or at the site of the disease, for example, in its tumor. In some embodiments, such a number or concentration of cells is detected in a subject for at least about 20 days, at least about 40 days, or at least about 60 days, or at least about 3 , 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or at least 2 or 3 years after administration of T lymphocytes, e.g., CAR-expressing T lymphocytes, and/or after administration of an immunomodulatory compound, such as lenalidomide. Such cell numbers can be detected using flow cytometric methods or quantitative PCR methods and extrapolation to total cell numbers using known methods See, for example, Brentjens et al., Sci Transl Med. 2013 5(177), Park et al, Molecular Therapy 15(4):825-833 (2007), Savoldo et al., JCI 121(5):1822-1826 (2011), Davila et al., (2013) PLoS ONE 8(4):e61338, Davila et al., Oncoimmunology 1(9):1577-1583 (2012), Lamers, Blood 2011 117:72-82, Jensen et al., Biol Blood Marrow Transplant 2010 September; 16(9): 1245-1256, Brentjens et al., Blood 2011 118(18):4817-4828.

[0575] In some aspects, the number of copies of the nucleic acid encoding the recombinant receptor, for example, the number of copies of the vector per 100 cells, for example, in peripheral blood or bone marrow or in another compartment, measured using immunohistochemistry, PCR and/or flow cytometry, is at least 0.01, at least 0.1, at least 1, or at least 10 after about 1 week after about 2 weeks after about 3 weeks after about 4 weeks , after about 5 weeks, or at least about 6 weeks, or at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or at least , 2 or 3 years after the introduction of cells, for example, CAR-expressing T-lymphocytes, and/or an immunomodulatory compound, for example, lenalidomide. In some embodiments, the number of copies of a vector expressing a receptor, e.g., CAR, per microgram of genomic DNA is at least 100, at least 1000, at least 5000 or at least 10000 or at least 15,000, or at least 20,000, at about 1 week, about 2 weeks, about 3 weeks, or at least about 4 weeks after administration of T lymphocytes, e.g., CAR-expressing T lymphocytes, or an immunomodulatory compound, e.g., lenalidomide, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or at least 2 or 3 years after such administration .

[0576] In some aspects, a receptor, e.g., CAR, expressed by cells is detected by quantitative PCR (qPCR) or by flow cytometry in the subject, in their plasma, serum, blood, tissue, and/or at the site of the disease, for example, at the site of the tumor, at a time point that is at least about 3 months, at least about 6 months, at least about 12 months, at least about 1 year, at least about 2 years , at least about 3 years, or more than 3 years, after administration of cells, for example, after the start of administration of T-lymphocytes, for example, CAR-expressing T-lymphocytes, and/or an immunomodulatory compound, for example, lenalidomide.

[0577] In some embodiments, the area under the curve (AUC) for the concentration of cells expressing a receptor (e.g., CAR-) in a fluid, plasma, serum, blood, tissue, organ, and/or disease site, e.g., in situ tumors in a subject, over time after administration of T-lymphocytes, e.g., CAR-expressing T-lymphocytes, and/or an immunomodulatory compound, e.g. lymphocytes, for example, CAR-expressing T-lymphocytes, without administration of an immunomodulatory compound.

[0578] In some aspects, the method results in high in vivo proliferation of the administered cells, such as measured by flow cytometry. In some aspects, high peak cell proportions are detected. For example, in some embodiments, at a peak or maximum level following administration of T lymphocytes, e.g., CAR expressing T lymphocytes, and/or an immunomodulatory compound, e.g., lenalidomide, in the blood, plasma, serum, tissue, or disease site of the subject , or in its leukocyte fraction, for example, in the PBMC fraction or in the T-lymphocyte fraction, at least about 10%, at least about 20%, at least about 30%, at least about 40% , at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% of the cells express the recombinant receptor, eg, CAR.

[0579] In some aspects, increasing or prolonging the proliferation and/or persistence of the dose of cells in a subject administered together with an immunomodulatory compound, eg, lenalidomide, is associated with improved tumor outcomes in the subject. In some embodiments, the outcome of treatment associated with the tumor includes a decrease in the mass of the tumor or a decrease in the number of bone marrow blast cells in the subject. In some embodiments, tumor weight is reduced by, or at least by, or about 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 percent after administration according to this method. In some embodiments, disease severity, tumor size, tumor volume, tumor masses, and/or tumor burden or bulk is reduced after cell dosing by at least or about 50%, 60%, 70%, 80% , 90% or more compared to a subject treated with a method that does not include the administration of an immunomodulatory compound, for example, lenalidomide.

B. Functional activity of T-lymphocytes

[0580] In some embodiments, parameters associated with therapy or treatment outcomes, which include parameters that can be assessed during the screening and/or evaluation of treatment outcomes and/or monitoring of treatment outcomes, include one or more parameters from activity, phenotype , proliferation or function of T-lymphocytes. In some embodiments, any assay known in the art can be used to evaluate the activity, phenotypes, proliferation, and/or function of T lymphocytes, eg, T lymphocytes administered for T cell therapy. Before and/or after administration of the cells and/or an immunomodulatory compound, eg, lenalidomide, the biological activity of the engineered cell populations is, in some embodiments, measured, for example, by a number of known methods. Parameters for evaluation include specific binding of engineered or natural T-lymphocytes or other immune cells to an antigen, in vivo, eg, by image processing, or ex vivo, eg, by ELISA or flow cytometry. In certain embodiments, the ability of engineered cells to destroy target cells can be measured using any appropriate method known in the art, such as cytotoxicity assays, such as those described in Kochenderfer et al., J. Immunotherapy, 32(7): 689- 702 (2009), and Herman et al., J. Immunological Methods, 285(1): 25-40 (2004). In certain embodiments, cell biological activity is measured by assaying the expression and/or secretion of one or more cytokines such as CD107a, IFNγ, IL-2, GM-CSF, and TNFα and/or by assessing cytolytic activity.

[0581] In some embodiments, assays for activity, phenotypes, proliferation, and/or function of T lymphocytes, e.g., T lymphocytes administered for T cell therapy, include, but are not limited to, ELISPOT, ELISA, cell proliferation , cytotoxic lymphocyte (CTL) assays, T-lymphocyte epitope, antigen or ligand binding or intracellular cytokine staining, proliferation assays, lymphokine secretion assays, direct cytotoxicity assays, and serial dilution assays. In some embodiments, proliferative responses of T lymphocytes can be measured, for example, by introducing ³ H-thymidine, BrdU (5-bromo-2'-deoxyuridine) or 2'-deoxy-5-ethynyluridine (EdU) into their DNA, or by dye dilution analysis using dyes such as carboxyfluorescein diacetate, succinimmunomodulatory compound ester (CFSE), CellTrace Violet, or PKH26 membrane dye.

[0582] In some embodiments, the assessment of the activity, phenotypes, proliferation and/or function of T lymphocytes, for example, T lymphocytes administered for T cell therapy, includes measuring cytokine production by T lymphocytes and/or measuring cytokine production in a biological a sample from a subject, for example, in plasma, serum, blood and/or tissue samples, for example, in tumor samples. In some cases, such measurable cytokines may include, without limitation, interleukin-2 (IL-2), interferon-gamma (IFNγ), interleukin-4 (IL-4), TNF-alpha (TNFα), interleukin-6 (IL -6), interleukin-10 (IL-10), interleukin-12 (IL-12), granulocyte macrophage colony stimulating factor (GM-CSF), CD107a and/or TGF-beta (TGFβ). Cytokine measurement assays are well known in the art and include, but are not limited to, ELISA, intracellular cytokine staining, bead cytometry, RT-PCR, ELISPOT, flow cytometry, and bioassays in which cells sensitive to the relevant cytokine are examined. susceptibility (eg proliferation) in the presence of the test sample.

[0583] In some embodiments, the assessment of the activity, phenotypes, proliferation and/or function of T lymphocytes, for example, T lymphocytes administered for T cell therapy, includes assessment of cell phenotypes, for example, the expression of specific cell surface markers. In some embodiments, T lymphocytes, eg, T lymphocytes administered for T cell therapy, are evaluated for expression of T lymphocyte activation markers, T lymphocyte depletion markers, and/or T lymphocyte differentiation markers. In some embodiments, the cell phenotype is assessed prior to administration. In some embodiments, the cell phenotype is assessed after administration. T-lymphocyte activation markers, T-lymphocyte depletion markers, and/or T-lymphocyte differentiation markers for evaluation include any markers known in the art for specific T-lymphocyte subsets, e.g., CD25, CD38, human leukocyte antigen DR (HLA-DR) , CD69, CD44, CD137, KLRG1, CD62L ^low , CCR7 ^low , CD71, CD2, CD54, CD58, CD244, CD160, programmed cell death protein 1 (PD-1), lymphocyte activation gene 3 protein (LAG-3), domain immunoglobulin of T-lymphocytes and protein mucin domain 3 (TIM-3), cytotoxic antigen of T-lymphocytes - protein 4 (CTLA-4), B-lymphocyte and T-lymphocyte attenuator (BTLA) and / or immunoglobulin of T-lymphocyte and inhibitory domain immunoreceptor tyrosine (TIGIT) motif (see, for example, Liu et al., Cell Death and Disease (2015) 6, e1792). In some embodiments, the cell surface marker being evaluated is CD25, PD-1, and/or TIM-3. In some embodiments, the cell surface marker being evaluated is CD25.

[0584] In some aspects, detecting expression levels includes performing an in vitro assay. In some embodiments, the in vitro assay is an immune assay, an aptamer-based assay, a histological or cytological assay, or an mRNA expression level assay. In some embodiments, the parameter or parameters for one or more of each one or more factors, effectors, enzymes, and/or surface markers are detected using an enzyme-linked immunosorbent assay (ELISA), immunoblot, immunoprecipitation, radioimmunoassay (RIA), immune staining, flow cytometry analysis, surface plasmon resonance (SPR), chemiluminescence analysis, horizontal flow immunoassay, inhibition assay, or avidity assay. In some embodiments, detection of cytokines and/or surface markers is determined using a binding reagent that specifically binds to at least one biomarker. In some cases, the binding reagent is an antibody or an antigen-binding fragment, an aptamer, or a nucleic acid probe.

[0585] In some embodiments, administration of an immunomodulatory compound, such as lenalidomide, increases the level of circulating CAR T lymphocytes.

C. Disease severity

[0586] In some embodiments, parameters associated with therapy or treatment outcomes, which include parameters that can be assessed during the screening and/or evaluation of treatment outcomes and/or monitoring of treatment outcomes, include tumor mass or disease severity. Administration of an immunotherapy, such as T cell therapy (eg, CAR expressing T lymphocytes) and/or an immunomodulatory compound, eg, lenalidomide, can reduce or prevent the spread or severity of a disease or condition in a subject. For example, when the disease or condition is a tumor, the methods generally reduce tumor size, bulk, metastasis, percentage of blast cells in the bone marrow, or molecularly detectable cancer, and/or improve prognosis or survival, or other associated symptom. with tumor mass.

[0587] In some embodiments, the proposed methods result in a reduction in tumor mass in treated subjects compared to alternative methods in which immunotherapy, such as T-cell therapy (e.g., CAR-expressing T-lymphocytes) is obtained without administering an immunomodulatory compound, such as lenalidomide. It is not necessary that tumor mass actually decrease in all subjects receiving combination therapy, but that tumor mass decrease on average in treated subjects, e.g. based on clinical data in which the majority of subjects treated with such combination therapy would demonstrate reduction in tumor mass, for example, at least 50%, 60%, 70%, 80%, 90%, 95% or more of subjects treated with combination therapy, showed a reduction in tumor mass.

[0588] Disease severity can encompass the total number of disease cells in a subject or in an organ, tissue, or bodily fluid of the subject, such as in a tumor organ or tissue, or elsewhere, such as that would indicate metastases. For example, tumor cells can be detected and/or quantified in blood, lymph, or bone marrow in the context of certain hematological malignancies. The severity of the disease may include, in some embodiments, the mass of the tumor, the number or extent of metastases, and/or the percentage of blast cells present in the bone marrow.

[0589] In some embodiments, the subject has myeloma, lymphoma, or leukemia. In some embodiments, the subject has non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), diffuse large B cell lymphoma (DLBCL), or a myeloma, such as multiple myeloma (MM). In some embodiments, the subject has MM or DBCBL. In some embodiments, the subject has follicular lymphoma (FL).

[0590] In some embodiments, the subject has a solid tumor.

[0591] In the case of MM, illustrative parameters for assessing the severity of the disease include parameters such as the number of clonal plasma cells (for example, > 10% in a bone marrow biopsy, or any number in a biopsy of other tissues; plasmacytoma), the presence of a monoclonal protein (paraprotein) either in serum or urine, evidence of a sense of end organ damage associated with a plasma cell disorder (eg, hypercalcemia (corrected calcium >2.75 mmol/L); renal failure attributed to myeloma; anemia (hemoglobin <10 g/dL), and / or bone damage (lytic lesions or osteoporosis with compression fractures)).

[0592] In the case of DLBCL, exemplary parameters for assessing disease severity include parameters such as cell morphology (e.g., centroblastic, immunoblastic, and anaplastic cells), gene expression, miRNA expression, and protein expression (e.g., expression of BCL2, BCL6, MUM1, LMO2 , MYC and p21).

[0593] In the case of leukemia, the severity of the disease can be determined by assessing residual leukemia in the blood or bone marrow. In some embodiments, the subject exhibits a morphological disorder if there are 5% or more blast cells in the bone marrow, such as as detected by light microscopy. In some embodiments, the subject demonstrates complete or clinical remission if there are less than 5% blast cells in the bone marrow.

[0594] In some embodiments, for leukemia, the subject may be in complete remission, but a small proportion of morphologically undetectable (by light microscopy technique) residual leukemic cells is present. A subject is said to exhibit minimal residual disease (MRD) if the subject exhibits less than 5% blast cells in the bone marrow and exhibits molecularly detectable cancer. In some embodiments, a molecularly detectable cancer may be assessed using any of a variety of molecular techniques that allow sensitive detection of a small number of cells. In some aspects, such techniques include PCR assays that can detect unique T-lymphocyte receptor Ig/gene rearrangements or fusion transcriptions produced by chromosomal translocations. In some embodiments, flow cytometry can be used to identify cancer cells based on leukemia-specific immunophenotypes. In some embodiments, molecular cancer detection can detect as little as 1 leukemia cell out of 100,000 normal cells. In some embodiments, the subject demonstrates an MRD that is molecularly detectable if at least 1 or more leukemia cells are detected among 100,000 cells, for example, by PCR or flow cytometry. In some embodiments, the severity of the disease in the subject is MRD ^- , or it is molecularly undetectable such that, in some cases, leukemia cells cannot be detected in the subject using PCR or flow cytometry techniques.

[0595] In some embodiments, the methods and/or administration of immunotherapy, such as T-cell therapy (e.g., CAR-expressing T-lymphocytes) and/or an immunomodulatory compound, e.g., lenalidomide, reduces disease severity relative to disease severity in a point in time just prior to the administration of the immunotherapy, eg, T cell therapy, and/or the immunomodulatory compound.

[0596] In some aspects, administration of an immunotherapy, eg, T cell therapy, and/or an immunomodulatory compound, eg, lenalidomide, may prevent an increase in disease severity, and this may be indicated by no change in disease severity.

[0597] In some embodiments, the method reduces the severity of the disease or condition, e.g., tumor cell count, tumor size, patient survival or event-free survival, to a greater extent and/or for a longer period of time than the reduction that was observed would be for a comparable method using an alternative therapy, such as one in which the subject receives immunotherapy, such as T cell therapy alone, without the administration of an immunomodulatory compound, such as lenalidomide. In some embodiments, the severity of the disease is reduced to a greater extent or for a longer time after combination therapy, with the introduction of immunotherapy, for example, T-cell therapy, and an immunomodulatory compound, for example, lenalidomide, compared with the reduction that would be carried out with the introduction each agent by itself, eg, when an immunomodulatory compound is administered to a subject not receiving immunotherapy, eg, T-cell therapy; or when administering immunotherapy, eg, T cell therapy, to a subject who is not receiving an immunomodulatory compound.

[0598] In some embodiments, the severity of a disease or condition in a subject is detected, assessed, or measured. The severity of a disease can be detected in some aspects by detecting the total number of diseased or disease-associated cells, such as tumor cells, in a subject, or in an organ, tissue, or bodily fluid of the subject, such as blood or serum. In some embodiments, disease severity, such as tumor weight, is assessed by measuring solid tumor weight and/or the number or extent of metastases. In some aspects, the survival of the subject, survival over a certain period of time, the degree of survival, the presence or duration of event-free or asymptomatic survival or survival without recurrence is assessed. In some embodiments, any symptom of a disease or condition is evaluated. In some embodiments, a measure of the severity of the disease or condition is specified. In some embodiments, exemplary parameters for determination include specific clinical results showing relief or improvement in a disease or condition, such as a tumor. Such parameters include: duration of disease control, including complete response (CR), partial response (PR), or stable disease (SD) (see e.g. Response Evaluation Criteria In Solid Tumors (RECIST) guidelines), tumor response as measured by objective evaluation ( ORR), progression-free survival (PFS), and overall survival (OS). Specific parameter thresholds may be selected to determine the efficacy of the combination therapy method provided herein.

[0599] In some embodiments, subjects treated with this method achieve a more robust response. In some cases, the measure of duration of response (DOR) includes the time from documenting tumor response to disease progression. In some embodiments, the response parameter may include a persistent response, such as a response that is maintained after a period of time has elapsed from the start of therapy. In some embodiments, a persistent response is indicated by a response rate at approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, or 24 months after the start of therapy. In some embodiments, the response is maintained for more than 3 months, more than 6 months, or more than 12 months. In some specific embodiments, subjects treated according to this method achieve a more robust response after a previous recurrence of the disease in the subject after remission in response to the administration of genetically engineered cells.

[0600] In some aspects, the severity of the disease is measured or detected before the administration of immunotherapy, for example, T-cell therapy, after the administration of immunotherapy, for example, T-cell therapy, but before the administration of an immunomodulatory compound, for example, lenalidomide, after the administration of an immunomodulatory compound, but prior to administration of immunotherapy, eg T cell therapy, and/or after administration of both immunotherapy, eg T cell therapy, and an immunomodulatory compound. In the context of multiple administration of one or more stages of combination therapy, the severity of the disease in some embodiments can be measured before or after the administration of any of the stages, doses and/or cycles of administration, or at the time point between the administration of any of the stages, doses and/or cycles of administration .

[0601] In some embodiments, the severity is reduced by or at least by, or about 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 percent using the proposed methods compared to the severity immediately prior to administration of an immunomodulatory compound, eg lenalidomide, and immunotherapy, eg T-cell therapy. In some embodiments, disease severity, tumor size, tumor volume, tumor mass, and/or tumor burden or bulk mass is reduced after administration of immunotherapy, e.g., T cell therapy, and an immunomodulatory compound by at least, or about 10 , 20, 30, 40, 50, 60, 70, 80, 90% or more compared with the parameters immediately before the introduction of immunotherapy, for example, T-cell therapy, and/or immunomodulatory compounds.

[0602] In some embodiments, reducing the severity of the disease by the method includes inducing a morphologically complete remission, for example, as assessed at 1 month, 2 months, 3 months, or more than 3 months after administration, for example, after the start of combination therapy.

[0603] In some aspects, the analysis of minimal residual disease, for example, as measured by multiparametric flow cytometry, is negative, or the level of minimal residual disease is less than about 0.3%, less than about 0.2%, less than about 0.1%. or less than about 0.05%.

[0604] In some embodiments, the event-free survival rate or overall survival rate of the subject is improved by these methods compared to other methods. For example, in some embodiments, the FFS or probability for subjects treated with the methods 6 months after the combination therapy method provided herein is greater than about 40%, greater than about 50%, greater than about than 60%, greater than about 70%, greater than about 80%, greater than about 90%, or greater than about 95%. In some aspects, the overall survival rate is greater than about 40%, greater than about 50%, greater than about 60%, greater than about 70%, greater than about 80%, greater than about 90%, or more than about 95%. In some embodiments, the subject treated with these methods demonstrates event-free survival, relapse-free survival, or survival for at least 6 months, or at least 1, 2, 3, 4, 5, 6 , 7, 8, 9 or 10 years. In some embodiments, the time to disease development is increased, e.g., the time to disease development is greater than or about greater than 6 months, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 years.

[0605] In some embodiments, after treatment with the method, the likelihood of recurrence of the disease is reduced compared to other methods. For example, in some embodiments, the likelihood of disease recurrence 6 months after application of the combination therapy method is less than about 80%, less than about 70%, less than about 60%, less than about 50%, less than about than 40%, less than about 30%, less than about 20%, or less than about 10%.

IV. Industrial products and kits

[0606] Industrial products are also provided containing an immunomodulatory drug (immunomodulatory compound), such as lenalidomide, and components for immunotherapy, for example, an antibody or antigen-binding fragment, or for T-cell therapy, for example, genetically engineered cells, and/or compositions thereof. The articles of manufacture may include a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, IV bags, and the like. Containers can be formed from a variety of materials such as glass or plastic. The container, in some embodiments, contains a composition that is contained alone or in combination with another composition effective for treating, preventing, and/or diagnosing a condition. In some embodiments, the container has a sterile access node. Exemplary containers include bags of intravenous solutions, vials, including vials with stoppers pierced by an injection needle, or bottles or vials for orally administered agents. A label or package insert may indicate that the composition is being used to treat a disease or condition.

[0607] An industrial product may contain (a) a first container with a composition contained therein, where the composition contains an antibody or genetically engineered cells used for immunotherapy, for example, T-cell therapy; and (b) a second container with a composition contained therein, wherein the composition contains a second agent such as an immunomodulatory compound, eg lenalidomide. The article of manufacture may further include a package insert indicating that the compositions can be used to treat a particular condition. Alternatively, or in addition, the article of manufacture may further comprise another or the same container containing a pharmaceutically acceptable buffer. In addition, it may contain other materials such as other buffers, diluents, filters, needles and/or syringes.

V Definitions

[0608] Unless otherwise specified, all art terms, designations, and other technical and scientific terms or terminology used herein are intended to have the same meaning as commonly understood by those skilled in the art to which the subject matter pertains. In some cases, terms with all intelligible meanings are defined herein for clarity and/or ease of reference, and the inclusion of such definitions in this document should not necessarily be construed as some significant difference from what is understood by everyone in the art.

[0609] As used herein, a "subject" is a mammal, such as a human or other animal, typically a human. In some embodiments, the subject, eg, the patient, to whom the immunomodulatory polypeptides, engineered cells, or compositions are administered, is a mammal, typically a primate, such as a human. In some embodiments, the primate is a different species of monkey. The subject may be male or female, and may be of any appropriate age, including children, adolescents, male or female, adult, and elderly subjects. In some embodiments, the subject is a mammal other than a primate, such as a rodent.

[0610] As used herein, “treatment” (and its grammatical variants such as “treat” or “treated”) refers to the total or partial alleviation or reduction of a disease or condition or disorder, or a symptom, adverse effect, or result, or a phenotype associated with it. The desired effects of treatment include, but are not limited to, preventing the occurrence or recurrence of the disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, preventing metastasis, reducing the rate of disease progression, ameliorating or alleviating the disease state, and remission or improving prognosis. The terms do not imply complete cure of a disease or complete elimination of any symptom or effect(s) on all symptoms or results.

[0611] As used herein, “slowing the progress of a disease” means delaying, hindering, slowing, delaying, stabilizing, suppressing, and/or stopping the development of a disease (such as cancer). This delay can be of varying lengths of time, depending on the history of the disease and/or from the disease being treated As is obvious, a significant or significant delay may result in preventing the disease from developing in an individual For example, a late stage cancer such as the development of metastases may be slowed down.

[0612] "Prevention", as used herein, includes the implementation of prevention regarding the occurrence or recurrence of a disease in a subject who may be predisposed to the disease, but in which the disease has not yet been diagnosed. In some embodiments, the provided cells and compositions are used to delay the development of a disease or to slow the progression of a disease.

[0613] As used herein, "suppression" of a function or activity is intended to reduce function or activity, compared to the same conditions, except for the condition or parameter of interest, or alternatively, compared to another condition. For example, cells that inhibit tumor growth reduce the rate of tumor growth compared to the rate of tumor growth in the absence of cells.

[0614] An “effective amount” of an agent, e.g., pharmaceutical, cell, or composition, in the context of administration, refers to an amount effective at doses/amounts and for periods of time necessary to achieve a desired result, such as a therapeutic or prophylactic result.

[0615] A “therapeutically effective amount” of an agent, such as a pharmaceutical drug or genetically engineered cells, refers to an amount effective at doses and for periods of time necessary to achieve a desired therapeutic result, for example, to treat a disease, condition, or disorder, and/ or for the pharmacokinetic or pharmacodynamic effects of the treatment. The therapeutically effective amount may vary according to such factors as the disease state, age, sex and body weight of the subject, and the immunomodulatory polypeptides or engineered cells administered. In some embodiments, the methods provided include administering the immunomodulatory polypeptides, engineered cells, or compositions at effective amounts, eg, at therapeutically effective amounts.

[0616] A “prophylactically effective amount” refers to an amount effective at dosages and for periods of time necessary to achieve the desired prophylactic result. Typically, but not necessarily, as long as a prophylactic dose is used in subjects before or at an early stage of disease, the prophylactically effective amount will be less than the therapeutically effective amount.

[0617] The term “pharmaceutical preparation” refers to a preparation that is in such a form as to allow effective biological activity of the active ingredient contained therein, and which does not contain additional components that are unacceptably toxic to the subject to whom the preparation would be administered.

[0618] A “pharmaceutically acceptable carrier” refers to an ingredient in a pharmaceutical preparation, other than the active ingredient, that is non-toxic to a subject. A pharmaceutically acceptable carrier includes, but is not limited to, a buffer, excipient, stabilizer, or preservative.

[0619] As used herein, reference to nucleotide or amino acid positions "corresponding" to nucleotide or amino acid positions in a described sequence, as given in the Sequence Listing, refers to the nucleotide or amino acid positions identified when aligned with the described sequence prior to being adjusted to maximum identity using a standard alignment algorithm such as the GAP algorithm. When aligning sequences, the corresponding residues can be identified, for example, using conserved and identical amino acid residues as criteria. As a general rule, to identify relevant positions, amino acid sequences are aligned to obtain the highest order of correspondence (see, for example: Computational Molecular Biology, Lesk, A.M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects , Smith, D.W., ed., Academic Press, New York, 1993;Computer Analysis of Sequence Data, Part I, Griffin, A.M., and Griffin, H.G., eds., Humana Press, New.Jersey, 1994;Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; Carrillo et al. (1988) SIAM J Applied Math 48:1073).

[0620] The term “vector”, as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes a vector as a self-replicating nucleic acid structure, as well as a vector introduced into the genome of the host cell into which it is introduced. Certain vectors are capable of directing the expression of the nucleic acids to which they are operably linked. Such vectors are referred to herein as "expression vectors". Among the vectors are viral vectors such as retroviral, eg gammaretroviral and lentiviral vectors.

[0621] The terms “host cell”, “host cell line”, and “host cell culture” are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells", which include the primary transformed cell and the progeny derived from it, regardless of the number of passages. The offspring may not be completely identical in nucleic acid content to the parent cell, but may contain mutations. Mutant progeny that have the same function or biological activity as after screening or selection for the original transformed cell are included herein.

[0622] As used herein, a statement that a cell or population of cells is "positive" for a particular marker refers to the detectable presence on or within a cell of a particular marker, typically a surface marker. When referring to a surface marker, the term refers to the presence of surface expression as detected by flow cytometry, for example, by staining with an antibody that specifically binds to the marker and detecting said antibody, where the staining can be detected by flow cytometry at a level substantially above the staining, detectable by the same procedure with an isotype-like control under otherwise identical conditions and/or at a level substantially similar to a cell known to be positive for the marker and/or at a level substantially higher than for a cell known to be negative for the marker. marker.

[0623] As used herein, a statement that a cell or population of cells is “negative” for a particular marker refers to the absence of a significant detectable presence on or in the cell of a particular marker, typically a surface marker. When referring to a surface marker, the term refers to the absence of surface expression as detected by flow cytometry, for example, by staining with an antibody that specifically binds to the marker and detecting said antibody, where the staining is not detectable by flow cytometry at a level significantly higher than staining. detectable by the same procedure using an isotype-like control under otherwise identical conditions, and/or detectable at a level substantially lower than for a cell known to be positive for the marker and/or at a level substantially similar compared to a cell known to be as negative relative to the marker.

[0624] Amino acid substitution may include replacing one amino acid in a polypeptide with another amino acid. The substitution may be a conservative amino acid substitution or a non-conservative amino acid substitution. Amino acid substitution can be introduced into the binding molecule, eg, the antibody of interest, and into products viewed for the desired activity, eg, preserved/improved antigen binding, reduced immunogenicity, or improved ADCC or CDC.

[0625] Amino acids can generally be grouped according to the following general side chain properties:

(1) hydrophobic: norleucine, Met, Ala, Val, Leu, Ile;

(2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;

(3) acidic: Asp, Glu;

(4) main: His, Lys, Arg;

(5) residues that affect chain orientation: Gly, Pro;

(6) aromatic: Trp, Tyr, Phe.

[0626] In some embodiments, conservative substitutions may include exchanging an element of one of these classes for another element of the same class. In some embodiments, non-conservative amino acid substitutions may include replacing an element from one of these classes with another class.

[0627] As used herein, "percent (%) amino acid sequence identity" and "percent identity" when used relative to an amino acid sequence (reference polypeptide sequence) is defined as the percentage of amino acid residues in a candidate sequence (e.g., in a subject's antibody or in its fragment) that are identical to amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percentage of sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways, for example using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Appropriate parameters for sequence alignment including any algorithms necessary to achieve maximum alignment over the entire length of the sequences being compared can be determined.

[0628] As used herein, singular forms include plural references unless the context clearly dictates otherwise. For example, referring to an item in the singular means "at least one" or "one or more". It is understood that aspects and variations described herein include aspects and variations of "comprising" and/or "consisting essentially of".

[0629] In the present description, various aspects of the claimed subject matter are presented in range format. It should be understood that the description in range format is for convenience and brevity only and should not be construed as an inflexible limitation on the scope of the claimed subject matter. Accordingly, a description of a range should be considered to include all possible subranges specifically described, as well as individual numerical values within those ranges. For example, when a range of values is given, it is understood that every intermediate value between the upper and lower limit of this range and any other stated or intermediate value in this stated range is covered by the claimed subject matter. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and they are also covered by the claimed subject matter, being subject to any specifically excluded limit in the stated range. When a stated range includes one or both of the limits, ranges excluding one or both of these included limits are also included in the claimed subject matter. This applies regardless of the range width.

[0630] The term "about" as used herein refers to the typical range of errors for the corresponding value, well known in the art. Reference to an “exemplary” value or parameter herein includes (and describes) embodiments that address that value or parameter per se. For example, a description referring to "about X" includes a description of "X".

[0631] As used herein, a composition refers to any mixture of two or more products, substances, or compounds, including cells. It may be a solution, suspension, liquid, powder, paste, aqueous, non-aqueous, or any combination thereof.

VI. Illustrative Embodiments

[0632] Among the proposed options for implementation are:

1. The method of treatment, the method includes:

(a) administering T cell therapy to a subject having a disease or condition; and

(b) administering to the subject an immunomodulatory compound.

2. The method of treatment, the method includes the introduction of T-cell therapy to a subject having a disease or condition, where at the time of the beginning of the introduction of T-cell therapy, the subject is administered, and / or he is being treated with it, an immunomodulatory compound, and / or a blood sample or a biopsy of the subject contains detectable levels of T-lymphocytes from therapy with genetically engineered T-lymphocytes.

3. The method of treatment, the method includes administering an immunomodulatory compound to a subject having a disease or condition, where at the time the administration of the immunomodulatory compound is started, the subject has previously been administered T-cell therapy for the treatment of the disease or condition, and/or a blood sample or a biopsy of the subject contains detectable levels T-lymphocytes from therapy with genetically engineered T-lymphocytes.

4. The method of any one of embodiments 1-4 and 28-34, wherein the method thereby prevents, reduces, or alleviates one or more symptoms or outcomes of diseases or conditions.

5. The method according to any of embodiments 1-4 and 28-34, where:

(a) the amount of the immunomodulatory compound administered is insufficient, as a single agent and/or without the administration of T cell therapy, to alleviate, reduce or prevent the disease or condition or symptom thereof, or result; and/or

(b) the amount of the immunomodulatory compound administered is insufficient, as a single agent and/or without the administration of T cell therapy, to alleviate, reduce or prevent the disease or condition in the subject or its symptom or result; and/or

(c) the method thereby reduces or alleviates the symptom or outcome or severity of the disease or condition to an extent that is greater than the combination of (i) the amount of reduction or alleviation achieved by administration of the immunomodulatory agent alone, optionally the average of the population of subjects having the disease or a condition, and (ii) the degree of reduction or amelioration by the administration of T cell therapy alone, optionally, the average for the population of subjects having the disease or condition; and/or

(d) the amount of the immunomodulatory compound administered in the method, or administered in one or more doses, is the maintenance dose of the compound or corresponds to the dose of the compound administered to subjects showing a response, optionally a complete response, following administration of the compound for treatment.

6. The method according to any one of embodiments 1-5 and 28-34, wherein the disease or condition is untreatable or resistant to the immunomodulatory compound and/or becomes untreatable or resistant thereto after treatment with the immunomodulatory compound; and/or the subject or disease or condition is determined to have a mutation or factor that causes the disease or condition to be resistant to treatment with an immunomodulatory compound.

7. The method of any one of embodiments 1-6 and 28-34, wherein the immunomodulatory compound is selected from the group consisting of: immunomodulatory drugs (IMiDs), thalidomide analogs, thalidomide derivatives, compounds that interact and/or bind to cereblon ( CRBN) and/or with one or more elements of the CRBN and E3 complex of ubiquitin ligase, Ikaros (IKZF1) inhibitors, Aiolos (IKZF3) inhibitors, compounds that enhance or facilitate ubiquitination and/or degradation of Ikaros (IKZF1) and/or Aiolos (IKZF3) .

8. The method of any one of embodiments 1-7 and 28-34, wherein administering the immunomodulatory compound comprises:

(i) at least one cycle greater than 30 days from the start of administration of the immunomodulatory compound, where the cycle includes administration of the compound, optionally daily, or at least daily, for up to 21 days, consecutively, and/or where the last administration of the compound in the cycle corresponds to 21 days or less after the first administration of the compound in the cycle; and/or

(ii) at least two cycles, every at least two cycles, comprising administering the compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or

(iii) administration, optionally daily, or at least daily, for no more than 14 consecutive days.

9. The method according to any of embodiments 1-8 and 28-34, where:

the start of administration of the immunomodulatory compound, or the start of administration of the compound in at least one cycle, and the start of administration of the T cell therapy occur on the same day or on consecutive days, optionally simultaneously; and/or

at least one dose of the immunomodulatory compound is administered on the same day, within one or two days, before or after a dose of T cell therapy.

10. The method of any one of embodiments 1 and 4-8, and 28-34, wherein the initiation of administration of the immunomodulatory compound, or the initiation of administration of the compound, in at least one cycle occurs prior to the initiation of administration of the T cell therapy.

11. The method of treatment, the method includes the introduction of T-cell therapy to a subject having a disease or condition, where the subject before the start of T-cell therapy is administered an immunomodulatory compound, where the cycle includes:

(i) administration for up to 21 days, consecutively, where the cycle includes more than 30 days from the start of administration of the immunomodulatory compound; and/or

(ii) administration for multiple consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or

(iii) administration for no more than 14 consecutive days.

12. The method of any one of embodiments 1, 2 and 4-11, and 28-34, wherein the start of administration of the immunomodulatory compound occurs within 14 days prior to the start of T cell therapy.

13. The method of any one of embodiments 1, 2 and 4-12, and 28-34 wherein administration of the immunomodulatory compound begins prior to administration of T cell therapy, beginning with:

(i) one week or within one week before or after collection from the subject of a sample containing T-lymphocytes to be processed and/or genetically engineered to receive therapy, optionally, where the sample is an apheresis sample; and/or

(ii) within 14 days prior to initiation of T cell therapy.

14. The method of any one of embodiments 1-13 and 28-34, wherein the T cell therapy comprises engineered cells to express the recombinant receptor.

15. The method of embodiment 14, wherein the genetic engineering comprises one or more ex vivo preparation method steps, optionally selected from:

(1) isolating cells from a biological sample using leukapheresis or apheresis;

(2) selecting or enriching cells using immunoaffinity methods;

(3) introducing a recombinant nucleic acid, optionally a viral vector, into the cells;

(4) incubating cells, optionally genetically engineered cells, in the presence of one or more stimulation conditions;

(5) preparing cells in the presence of a cryoprotectant; and/or

(6) preparing cells for administration to a subject, optionally in the presence of a pharmaceutically acceptable excipient.

(6) preparing cells for administration to a subject, optionally in the presence of a pharmaceutically acceptable excipient.

16. The method of embodiment 14 or 15, further comprising performing a method of preparing and/or further genetically engineering T lymphocytes to express the recombinant receptor, thereby generating a T cell therapy.

17. The method of embodiment 16 further comprising contacting the cells with the immunomodulatory compound during one or more steps of the ex vivo preparation method.

18. The method of any one of embodiments 1-16 and 28-34, wherein the T cell therapy comprises engineered T lymphocytes obtained by a preparation method comprising ex vivo incubation of the cells in the presence of an immunomodulatory compound.

19. The method of Embodiment 17 or Embodiment 18, wherein the incubation of the cells in the presence of one or more stimulus conditions is carried out in the presence of an immunomodulatory compound.

20. The method of any one of embodiments 1, 2 and 4-19, and 28-34, wherein the start of administration of the immunomodulatory compound occurs within 10 days, 7 days, 4 days, 3 days, or 2 days prior to the start of administration of T cell therapy .

21. The method of Embodiment 1, wherein the start of administration of the immunomodulatory compound in at least one cycle occurs after the start of administration of T cell therapy.

22. A method of treatment, the method includes administering an immunomodulatory compound to a subject, the subject has a disease or condition and is administered T-cell therapy, where the immunomodulatory compound is administered in a cycle including:

(i) administration of the immunomodulatory compound for up to 21 days, consecutively, where the cycle includes more than 30 days from the start of administration of the immunomodulatory compound; and/or

(ii) administering the immunomodulatory compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or

(iii) administering the immunomodulatory compound for no more than 14 days, consecutively.

23. The method of any one of embodiments 1-22, wherein the T cell therapy is such that the peak population of cells from the therapy, which are optionally CD3+ or CD8+ cells from the T cell therapy and/or optionally are T-lymphocytes CAR+, in blood is ((a) the mean of a plurality of subjects treated with T-cell therapy without administration of an immunomodulatory compound, or (b) subjects after administration of T-cell therapy) less than 10 cells per µl, less 5 cells per µl or less than 1 cell per µl.

24. The method of any one of embodiments 1-23, wherein the T cell therapy comprises cells expressing a recombinant receptor, optionally a CAR.

25. The method of embodiment 24, wherein the recombinant receptor comprises an antigen-binding domain specific for B-cell maturation antigen (BCMA).

26. The method of embodiment 1, wherein the start of administration of the immunomodulatory compound in at least one cycle is after the start of administration of T cell therapy.

27. The method according to any one of embodiments 1 and 3-26 and 28-34, wherein the start of administration of the immunomodulatory compound is at least 2 days later, at least 1 week later, at least 2 weeks later, at least 3 weeks or at least 4 weeks after the start of administration or after the last dose of T-cell therapy, and / or carried out after 2-28 days or 7-21 days after the start of administration or after the last dose of T -cell therapy.

28. The method of treatment, the method includes:

(a) administering T cell therapy to a subject having a disease or condition, and

(b) administering to the subject an immunomodulatory compound, wherein the start of administration of the immunomodulatory compound corresponds to the time point:

(1) at least 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after starting T cell therapy , and/or it is carried out after 2-28 or 7-21 days after the start of the introduction of T-cell therapy; and/or

(2) at or after the time point, optionally immediately thereafter or within 1-3 days after: (i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

29. The method of treatment, the method includes administering an immunomodulatory compound to a subject who, prior to the start of the administration of the immunomodulatory compound, is administered T-cell therapy for the treatment of a disease or condition, where the start of the administration of the immunomodulatory compound corresponds to the time point:

(1) at least 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after initiation of T cell therapy , and/or it is carried out after 2-28 days or 7-21 days after the start of the introduction of T-cell therapy; and/or

(2) at or after the time point, optionally immediately thereafter or within 1-3 days after: (i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

30. The method of any one of embodiments 26-29, wherein the start of administration of the immunomodulatory compound is after a period of time that is greater than about 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 24 days, or 28 days after the start of T-cell therapy.

31. The method of any one of embodiments 26-30, comprising, prior to initiating administration of the immunomodulatory compound, selecting a subject in which: (i) a peak maximum cell level from T-cell therapy is detected in the blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

32. A method of treatment comprising administering a therapeutically effective amount of an immunomodulatory compound to a subject who, prior to initiation of administration of the immunomodulatory compound, is administered T cell therapy for the treatment of a disease or condition, wherein the subject is one that on or about day 12-15, optionally , on or about day 14 after initiation of T-cell therapy for the treatment of a disease or condition:

(i) the number of cells from T-cell therapy in the subject is less than 75% of the average number of cells from T-cell therapy at the same time in a plurality of subjects who are administered the same or similar dose of T-cell therapy; and/or

(ii) the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, in the blood is less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl.

33. Method of treatment, including:

(a) selecting a subject that corresponds to or approximately corresponds to day 12-15, optionally, day 14 or so, after the start of the administration of T cell therapy for the treatment of the disease or condition:

(i) the number of cells from T-cell therapy in the subject is less than 75% of the average number of cells from T-cell therapy at the same time in a plurality of subjects who are administered the same or similar dose of T-cell therapy; and/or

(ii) the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, in the blood is less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl; and

(b) administering a therapeutically effective amount of the immunomodulatory compound to the subject.

34. The method of any one of embodiments 33 wherein the immunomodulatory compound is administered daily, optionally once daily.

35. The method according to any of embodiments 1-34, wherein the immunomodulatory compound is administered for more than or about more than 7 consecutive days, more than or about more than 14 consecutive days, more than or about more than 21 consecutive days, or more than or more than about 28 consecutive days.

36. The method of any one of embodiments 1-35, wherein the immunomodulatory compound is administered in a cycle comprising administration daily for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered.

37. The method of embodiment 36, wherein the rest period during which no immunomodulatory compound is administered is greater than 7 consecutive days, greater than 14 consecutive days, greater than 21 days, or greater than 28 days.

38. The method of any one of embodiments 1-15, 11-16, 25 and 26, wherein the cycle of administration of the immunomodulatory compound is repeated at least once.

39. The method of any one of embodiments 1-39, wherein the immunomodulatory compound is administered for at least 2 cycles, at least 3 cycles, at least 4 cycles, at least 5 cycles, at least , 6 cycles, at least 7 cycles, at least 8 cycles, at least 9 cycles, at least 10 cycles, at least 11 cycles, or at least 12 cycles.

40. The method of any one of embodiments 1-39, wherein the administration of the immunomodulatory compound continues at least from the time after the start of administration of the T-lymphocytes until:

the number of cells from, or derived from, the administered T cell therapy detectable in the blood of the subject will not increase compared to the number of cells in the subject at the time immediately prior to the administration of the immunomodulatory compound or compared to the time prior to the administration of the T cell therapy;

the number of cells from, or derived from, T-cell therapy detectable in the blood will not be within 2.0 times (more or less) the peak or maximum number observed in the subject's blood after initiation of T-lymphocyte administration;

the number of cells from T-cell therapy detected in the subject's blood will not become more than or about more than 10%, 15%, 20%, 30%, 40%, 50%, or 60% of the total number of peripheral blood mononuclear cells (PBMC) in the subject's blood; and/or

the subject will not show a reduction in tumor weight compared to tumor weight at the time immediately prior to administration of the T cell therapy or at the time immediately prior to administration of the immunomodulatory compound; and/or

the subject will not demonstrate complete or clinical remission.

41. The method of any one of embodiments 1-40, wherein the immunomodulatory compound binds to cereblon (CRBN) and/or a complex of CRBN and E3 ubiquitin ligase; and/or inhibits the growth transcription factor Ikaros (IKZF1) or Aiolos (IKZF3); and/or enhances ubiquitination or degradation of Ikaros (IKZF1) or Aiolos (IKZF3).

42. The method of any one of embodiments 1-41, wherein the immunomodulatory compound is thalidomide or is a derivative or analogue of thalidomide.

43. The method of any one of embodiments 1-42, wherein the immunomodulatory compound is lenalidomide, pomalidomide, avadomide, a stereoisomer of lenalidomide, pomalidomide, avadomide, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, clathrate, or polymorph thereof.

44. The method according to any one of embodiments 1-43, wherein the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione, its stereoisomer or enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

45. The method of any one of embodiments 1-44, wherein the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione.

46. The method of any one of embodiments 1-43, wherein the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione, its stereoisomer or enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

47. The method of any of embodiments 1-43 and 46 wherein the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione .

48. The method of any one of embodiments 1-47, wherein the immunomodulatory compound is administered orally, subcutaneously, or intravenously.

49. The method of embodiment 46 wherein the immunomodulatory compound is administered orally.

50. The method of any one of embodiments 1-48, wherein the immunomodulatory compound is administered in a capsule or tablet.

51. The method of any one of embodiments 1-50, wherein the immunomodulatory compound is administered in an amount of from or about 0.1 mg to about 100 mg, from or about 0.1 mg to 50 mg, from or about 0.1 mg to 25 mg, from or about 0.1 mg to 10 mg, from or about 0.1 mg to 5 mg, from or about 0.1 mg to 1 mg, from or about 1 mg to 100 mg , from or about 1 mg to 50 mg, from or about 1 mg to 25 mg, from or about 1 mg to 10 mg, from or about 1 mg to 5 mg, from or about 5 mg to 100 mg , from or about 5 mg to 50 mg, from or about 5 mg to 25 mg, from or about 5 mg to 10 mg, from or about 10 mg to 100 mg, from or about 10 mg to 50 mg , from or from 10 mg to 25 mg, from or from about 25 mg to 100 mg, from or from about 25 mg to 50 mg, or from or from about 50 mg to 100 mg, each inclusive.

52. The method of any one of embodiments 1-51, wherein the immunomodulatory compound is administered once a day, twice a day, three times a day, four times a day, five times a day, or six times a day.

53. The method of any one of embodiments 1-52, wherein the immunomodulatory compound is administered at a total daily dosage amount of at least, or about, at least 0.1 mg per day, 0.5 mg per day, 1.0 mg per day, 2.5 mg per day, 5 mg per day, 10 mg per day, 25 mg per day, 50 mg per day, or 100 mg per day.

54. The method according to any one of embodiments 1-53, where:

the immunomodulatory compound is administered in an amount greater than or about greater than 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg, and less than 25 mg; or

the immunomodulatory compound is administered in an amount greater than or about greater than 1 mg per day, 2.5 mg per day, 5 mg per day, 7.5 mg per day, 10 mg per day, 15 mg per day, and less than 25 mg per day .

55. The method of any one of embodiments 1-54, wherein administration of a therapeutically effective amount of the immunomodulatory compound stimulates an increase in proliferation of T-lymphocytes associated with T cell therapy compared to proliferation following administration of T cell therapy without the immunomodulatory compound.

56. The method of any one of embodiments 1-55, wherein administration of a therapeutically effective amount of the immunomodulatory compound stimulates an increase in cytolytic activity mediated by T lymphocytes associated with T cell therapy compared to cytolytic activity following administration of T lymphocytes without the immunomodulatory compound.

57. The method of any one of embodiments 1-56, wherein administration of a therapeutically effective amount of the immunomodulatory compound stimulates an increase in cytokine production by T lymphocytes associated with T cell therapy compared to cytokine production following administration of T lymphocytes without the immunomodulatory compound.

58. The method of any one of embodiments 55-57 wherein the magnification is greater than or about greater than 1.5x, 2.0x, 3.0x, 4.0x, 5.0x, 10, 0 times or more.

59. The method of any one of embodiments 1-58, wherein the T cell therapy is or comprises tumor infiltrating lymphocyte (TIL) therapy or engineered cells expressing a recombinant receptor that specifically binds to an antigen.

60. The method of any one of embodiments 1-59, wherein the T cell therapy is or comprises engineered cells expressing a recombinant receptor that specifically binds to an antigen.

61. The method of any one of embodiments 1-60, wherein the T cell therapy comprises cells expressing a recombinant receptor that is or contains a functional non-TCR antigen receptor or a TCR or antigen-binding fragment thereof.

62. The method of embodiment 61 wherein the recombinant antigen receptor is a chimeric antigen receptor (CAR).

63. The method of any one of embodiments 1-62, wherein the T cell therapy comprises a recombinant antigen receptor that contains an extracellular domain containing an antigen-binding domain that specifically binds to an antigen.

64. The method of any one of embodiments 62 or 63, wherein the antigen is associated with, specific to, and/or expressed in a cell or tissue of the disease, disorder, or condition.

65. The method of embodiment 64, wherein the disease, disorder, or condition is an infectious disease or disorder, an autoimmune disease, an inflammatory disease, or a tumor or cancer.

66. The method of any one of embodiments 62-65, wherein the antigen is a tumor antigen.

67. The method of any of embodiments 62-66 wherein the antigen is selected from ROR1, B cell maturation antigen (BCMA), charcoal anhydrase 9 (CAIX), tEGFR, Her2/neu (erbB2 tyrosine kinase receptor), L1-CAM, CD19 , CD20, CD22, mesothelin, CEA and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40) , EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate-binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha , IL-13R-alpha2, kinase insert domain-containing receptor (kdr), kappa light chain, Ley antigen, L1 cell adhesion molecule, (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, preferably expressed melanoma antigen (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE Al, HLA-A2 NY-ESO-1, PSCA , folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, testicular cancer antigen, mesothelin, mouse CMV, mucin 1 (MUC1) , MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, G protein coupled receptor 5D (GPCR5D), oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu , estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms tumor 1 (WT-1), cyclin, cyclin A2, CCL-1, CD138, optionally, a human antigen to any of the above; a pathogen-specific antigen; and an antigen associated with a universal label.

68. The method of any one of embodiments 62-67, wherein the antigen is or comprises CD19, optionally human CD19.

69. The method of any one of embodiments 62-68, wherein the antigen is or comprises an antigen associated with multiple myeloma, optionally BCMA, optionally human BCMA.

70. The method of any one of embodiments 62-69, wherein the antigen binding domain is or contains an antibody or antibody fragment, which is optionally a single chain fragment.

71. The method of embodiment 70, wherein the fragment comprises antibody variable regions connected by a flexible linker.

72. The method of embodiment 70 or embodiment 71, wherein the fragment contains scFv.

73. The method of any one of embodiments 62-72, wherein the T cell therapy comprises a recombinant receptor that further comprises a spacer, optionally derived from an immunoglobulin, optionally containing a hinge region.

74. The method of any one of embodiments 62-73, wherein the recombinant antigen receptor comprises an intracellular signaling region.

75. The method of embodiment 74 wherein the intracellular signaling region comprises an intracellular signaling domain.

76. The method of embodiment 75, wherein the intracellular signaling domain is or comprises a primary signaling domain, which is a signaling domain that can induce a primary activation signal in a T-lymphocyte, a component of the T-lymphocyte receptor (TCR) signaling domain, and/or a signaling domain a domain containing an immunoreceptor tyrosine activating motif (ITAM).

77. The method of Embodiment 75 or Embodiment 76, wherein the intracellular signaling domain is or comprises an intracellular signaling domain of the CD3 chain, optionally the CD3-zeta (CD3ζ) chain, or a signal portion thereof.

78. The method of any one of embodiments 75-77, wherein the recombinant receptor further comprises a transmembrane domain located between an extracellular domain and an intracellular signaling region, wherein the transmembrane domain is optionally a CD8 or CD28 transmembrane domain.

79. The method of any one of embodiments 75-78, wherein the intracellular signaling region further comprises a costimulatory signaling region.

80. The method of embodiment 79, wherein the co-stimulatory signaling region comprises an intracellular signaling domain of a co-stimulatory T-lymphocyte molecule or a signaling portion thereof.

81. The method of embodiment 79 or embodiment 80, wherein the costimulatory signaling region comprises a CD28, 4-1BB, or ICOS intracellular signaling domain, or a signaling portion thereof.

82. The method of any one of embodiments 79-81, wherein the co-stimulatory signaling region comprises a 4-1BB intracellular signaling domain.

83. The method of any one of embodiments 79-82, wherein the co-stimulatory signaling region is between the transmembrane domain and the intracellular signaling region.

84. The method of any one of embodiments 1-83, wherein the T cell therapy comprises:

T-lymphocytes selected from the group consisting of central memory T-lymphocytes, effector memory T-lymphocytes, intact T-lymphocytes, stem central memory T-lymphocytes, effector T-lymphocytes and regulatory T-lymphocytes; and/or

a plurality of cells, this plurality contains at least 50% of a population of cells selected from the group consisting of CD4+ T lymphocytes, CD8+ T lymphocytes, central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central memory T-lymphocytes, effector T-lymphocytes and regulatory T-lymphocytes.

85. The method of any one of embodiments 1-84, wherein the T cell therapy comprises T lymphocytes that are CD4+ or CD8+.

86. The method of any one of embodiments 1-85, wherein the T cell therapy comprises primary cells obtained from a subject.

87. The method of any one of embodiments 1-86, wherein the T cell therapy comprises cells that are autologous to the subject.

88. The method of any one of embodiments 1-87, wherein the T cell therapy comprises T lymphocytes that are allogeneic to the subject.

89. The method of any one of embodiments 1-88, wherein the subject is a human.

90. The method of any one of embodiments 1-89, wherein the T cell therapy comprises administering from or about 1 x 10 ⁵ to 1 x 10 ⁸ cells, in total, T lymphocytes expressing recombinant receptors, in total, or mononuclear cells peripheral blood cells (PBMC), in total, from or about 5 × 10 ⁵ to 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, or from or about 1 x 10 ⁶ to 1 x 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, each value is inclusive.

91. The method of any one of embodiments 1-90, wherein the T cell therapy comprises administering no more than 1 x 10 ⁸ cells, in total, of T lymphocytes expressing recombinant receptors in total, or peripheral blood mononuclear cells (PBMCs), in total. not more than 1 × 10 ⁷ cells in total, recombinant receptor-expressing T-lymphocytes in total, or peripheral blood mononuclear cells (PBMCs) in total, not more than 0.5 × 10 ⁷ cells in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMCs), in total, not more than 1 × 10 ⁶ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells ( PBMC), in total, not more than 0.5 × 10 ⁶ cells, in total, T-lymphocytes expressing recombinant receptors, in general, or peripheral blood mononuclear cells (PBMC), in general.

92. The method of any one of embodiments 1-91, wherein the number of cells administered in the T cell therapy is less than that in the other method, wherein the T cell therapy is administered without the administration of an immunomodulatory compound, optionally, this other method yields resulting in a similar or lower degree of alleviation or reduction or prevention of the disease or condition or symptom, or their severity, compared to the result of the present method.

93. The method of embodiment 92, wherein the number of cells administered is 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, or 10-fold less than the amount administered in the other method.

94. The method of any one of embodiments 1-93, wherein the T cell therapy is administered as a single pharmaceutical composition containing cells.

95. The method of any one of embodiments 1-94, wherein the T cell therapy comprises a cell dose that is a divided dose, where the cells of the dose are administered in a plurality of compositions collectively containing all the cells of the dose, over a period of no more than three days.

96. The method of any one of embodiments 1-95, wherein the method further comprises administering anti-lymphocyte chemotherapy prior to administering T cell therapy.

97. The method of any one of embodiments 1-96, wherein the disease or condition is a cancer.

98. The method of any one of embodiments 1-97, wherein the cancer is B-lymphocyte cancer and/or myeloma, lymphoma, or leukemia.

99. The method of embodiment 97 or embodiment 98 wherein the cancer is mantle cell lymphoma (MCL), multiple myeloma (MM), acute lymphocytic leukemia (ALL), adult ALL, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma ( NHL) or diffuse large B-cell lymphoma (DLBCL).

100. The method of Embodiment 97 wherein the cancer is a non-hematologic cancer or is a solid tumor.

101. The method of any one of embodiments 1-100, wherein the T cell therapy exhibits an increase or prolongation of proliferation and/or persistence in a subject compared to a method in which the T cell therapy is administered to the subject without an immunomodulatory compound.

102. The method of any one of embodiments 1-101, wherein the method reduces tumor mass to a greater extent and/or for a longer period of time compared to the reduction that would be observed for a comparable method in which T cell therapy is administered to a subject without immunomodulatory compounds and/or in which the immunomodulatory compound is administered without T cell therapy, optionally at the same dose or time schedule of dosing.

103. A set containing:

(a) a pharmaceutical composition containing a standard unit dose of T-cell therapy; and

(b) instructions for administering the composition to a subject having a disease or condition in conjunction with administering a composition containing an immunomodulatory compound, wherein the instructions specify administering the immunomodulatory compound in one or more unit dose units according to an administration cycle including:

(i) administration of the immunomodulatory compound for up to 21 days, consecutively, where the cycle includes more than 30 days from the start of administration of the immunomodulatory compound; and/or

(ii) administering the immunomodulatory compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or

(iii) administering the immunomodulatory compound for no more than 14 days, consecutively.

104. A set containing:

(a) a pharmaceutical composition containing one or more standard unit doses of an immunomodulatory compound; and

(b) instructions for administering an immunomodulatory compound to a subject having a disease or condition in conjunction with administering a unit dose of a pharmaceutical composition comprising a T cell therapy, wherein the instructions specify the administration of one or more unit dose units of the immunomodulatory compound according to an administration cycle comprising:

(i) administration of the immunomodulatory compound for up to 21 days, consecutively, where the cycle includes more than 30 days from the start of administration of the immunomodulatory compound; and/or

(ii) administering the immunomodulatory compound for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered, where the rest period is greater than 14 consecutive days; and/or

(iii) administering the immunomodulatory compound for no more than 14 days, consecutively.

105. The kit of embodiment 103 or embodiment 104, wherein the instructions specify the initiation of administration of one or more standard unit doses of the immunomodulatory compound on the same day, optionally at the same time as the initiation of administration of T cell therapy.

106. The kit of embodiment 103 or embodiment 104, wherein the instructions specify the initiation of administration of one or more standard unit doses of the immunomodulatory compound prior to the initiation of administration of T cell therapy.

107. The kit of embodiment 106, wherein the instructions specify the start of administration of one or more standard unit doses of an immunomodulatory compound:

(1) a week or within a week prior to collection from the subject of a sample containing T-lymphocytes to be genetically engineered, optionally, where the sample is an apheresis sample; and/or

(2) during one or more of the steps of an ex vivo preparation method for receiving therapy with genetically engineered T-lymphocytes; and/or

(3) within 14 days prior to the introduction of T-cell therapy.

108. The kit of embodiment 107 wherein one or more steps of the ex vivo preparation method are selected from:

(1) isolating cells from a biological sample using leukapheresis or apheresis;

(2) selection or enrichment of cells using immunoaffinity methods;

(3) introducing the recombinant nucleic acid, optionally a viral vector, into the cells;

(4) incubating cells, optionally genetically engineered cells, in the presence of one or more stimulation conditions;

(5) preparing cells in the presence of a cryoprotectant; and/or

(6) preparing cells for administration to a subject, optionally in the presence of a pharmaceutically acceptable excipient.

109. The kit of any one of embodiments 103-106, wherein the instructions specify the initiation of administration of one or more standard unit doses of the immunomodulatory compound within 10 days, 7 days, 4 days, 3 days, or 2 days prior to the administration of T cell therapy.

110. The kit of embodiment 103 or embodiment 104, wherein the instructions specify the initiation of administration of one or more standard unit doses of the immunomodulatory compound after initiation of administration of T cell therapy.

111. The kit of embodiment 110, wherein the instructions specify the start of administration of one or more standard unit doses of the immunomodulatory compound in at least 2 days, at least 1 week, at least 2 weeks, at least 3 weeks or at least 4 weeks after the start of the administration of T-cell therapy, and/or 2-28 days or 7-21 days after the start of the administration of T-cell therapy.

112. A set containing:

(a) a pharmaceutical composition containing a standard unit dose of T cell therapy and

(b) instructions for administering the composition to a subject having a disease or condition in conjunction with administering an immunomodulatory compound, wherein the instructions specify the start of administration of the immunomodulatory compound at one or more unit doses at a point in time:

(1) at least 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after starting T cell therapy , and/or it is carried out after 2-28 days or 7-21 days after the start of the introduction of T-cell therapy; and/or

(2) at or after the time point, optionally immediately thereafter or within 1-3 days after: (i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

113. A set containing:

(a) a pharmaceutical composition containing one or more standard unit doses of an immunomodulatory compound; and

(b) instructions for administering an immunomodulatory compound to a subject having a disease or condition in conjunction with administering a unit dose of a pharmaceutical composition comprising a T cell therapy, wherein the instructions specify the start of administration of one or more unit dose units of the immunomodulatory compound at a time point:

(1) at least 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after starting T cell therapy , and/or it is carried out after 2-28 days or 7-21 days after the start of the introduction of T-cell therapy; and/or

(2) at or after the time point, optionally immediately thereafter or within 1-3 days after: (i) a peak maximum cell level from T-cell therapy is detected in the subject's blood; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

114. The kit of embodiment 112 or embodiment 113, wherein the instructions specify the commencement of administration of one or more standard unit doses of the immunomodulatory compound after a period of time that is greater than or greater than about 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 24 days, or 28 days after the start of T-cell therapy.

115. The kit of any one of embodiments 112-114, wherein the instructions specify the selection of a subject for administration of one or more standard unit doses of an immunomodulatory compound following administration of T cell therapy, wherein: (i) a peak maximum cell level from T is detected in the subject's blood - cell therapy; (ii) the number of cells from T-cell therapy detectable in the blood, after their detection in the blood, becomes undetectable or decreases, optionally, decreases compared with the previous time point after the introduction of T-cell therapy; (iii) the number of cells from T-cell therapy detected in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more compared to the peak or maximum number of cells from T-cell therapy detected in the blood of the subject, after the start of the introduction of T-cell therapy; (iv) at some point in time after detecting the peak or maximum level of cells from T-cell therapy in the blood of the subject, the number of T-lymphocytes or cells derived from them, detected in the blood of the subject, is less than 10%, less than 5%, less than 1% or less than 0.1% of the total peripheral blood mononuclear cells (PBMC) in the subject's blood; (v) the subject demonstrates the development of the disease and/or its recurrence after remission after treatment with T-cell therapy; and/or (vi) the subject exhibits an increase in tumor weight relative to tumor weight at the time point before or after administration of the T-lymphocytes and prior to the start of administration of the immunomodulatory compound.

116. A set containing:

(a) a pharmaceutical composition containing a standard unit dose of T-cell therapy; and

(b) instructions for administering the composition to a subject having a disease or condition in conjunction with administering an immunomodulatory compound, wherein the instructions specify administering the immunomodulatory compound to a subject in one or more unit doses if on or about day 12-15, optionally, day 14 or so, after initiation of T-cell therapy for the treatment of a disease or condition:

(i) the number of cells from T-cell therapy in the subject is less than 75% of the average number of cells from T-cell therapy at the same time in a plurality of subjects who are administered the same or similar dose of T-cell therapy; and/or

(ii) the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, in the blood is less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl.

117. A set containing:

(a) a pharmaceutical composition containing one or more standard unit doses of an immunomodulatory compound; and

(b) instructions for administering one or more unit dose units of an immunomodulatory compound to a subject having a disease or condition in conjunction with administering a pharmaceutical composition comprising a unit dose of a T cell therapy, wherein the instructions specify administering one or more unit unit doses of an immunomodulatory compound to a subject if on or about day 12-15, optionally on or about day 14, after initiation of T-cell therapy for the treatment of a disease or condition:

(i) the number of cells from T-cell therapy in the subject is less than 75% of the average number of cells from T-cell therapy at the same time in a plurality of subjects who are administered the same or similar dose of T-cell therapy; and/or

(ii) the number of CD3+ or CD8+ cells from T cell therapy, optionally CAR+ T lymphocytes, in the blood is less than 10 cells per μl, less than 5 cells per μl, or less than 1 cell per μl.

118. The kit of any one of embodiments 103-117, wherein the immunomodulatory compound is formulated in an amount for daily administration and/or instructions specify daily administration of the immunomodulatory compound.

119. The kit of any one of embodiments 103-118, wherein the instructions specify administering the immunomodulatory compound for more than or about more than 7 consecutive days, more than or about more than 14 consecutive days, more than or about more than 21 consecutive days, more than or about more than 21 consecutive days. days or more, or about more than 28 consecutive days.

120. The kit of any of embodiments 103-119, wherein the instructions specify the administration of the immunomodulatory compound in a cycle of administration, including daily administration for a plurality of consecutive days followed by a rest period during which no immunomodulatory compound is administered.

121. The kit of embodiment 120, wherein the instructions specify a rest period during which no immunomodulatory compound is administered, greater than 7 consecutive days, greater than 14 consecutive days, greater than 21 days, or greater than 28 days.

122. The kit of any one of embodiments 103-121, wherein the instructions specify a cycle of administration of the immunomodulatory compound that is repeated at least once.

123. The kit of any one of embodiments 103-122, wherein the instructions specify the continuation of the administration of the immunomodulatory compound, at least from the time after the start of the administration of T-lymphocytes, until:

the number of cells from or derived from the administered T-cell therapy detectable in the blood of the subject will not increase compared to the number of cells of the subject at the time immediately before the administration of the immunomodulatory compound or compared with the previous time after the administration of the T-cell therapy;

the number of cells from or derived from T-cell therapy detected in the blood will not be within 2.0 times (more or less) the peak or maximum number observed in the blood of the subject after the start of the administration of T-lymphocytes;

the number of cells from T-cell therapy detected in the subject's blood will not become more than or about more than 10%, 15%, 20%, 30%, 40%, 50%, or 60% of the total number of peripheral blood mononuclear cells (PBMC) in the subject's blood; and/or

the subject will not show a reduction in tumor weight compared to tumor weight at the time immediately prior to administration of the T cell therapy or at the time immediately prior to administration of the immunomodulatory compound; and/or

the subject will not demonstrate complete or clinical remission.

124. A kit according to any one of embodiments 103-123, wherein the immunomodulatory compound binds to cereblon (CRBN) and/or a complex of CRBN and E3 ubiquitin ligase; and/or inhibits the transcription factor Ikaros (IKZF1) or Aiolos (IKZF3); and/or enhances ubiquitination or degradation of Ikaros (IKZF1) or Aiolos (IKZF3).

125. The kit of any one of embodiments 103-124, wherein the immunomodulatory compound is thalidomide or is a derivative or analog of thalidomide.

126. The kit of any one of embodiments 103-125, wherein the immunomodulatory compound is lenalidomide, pomalidomide, avadomide, a stereoisomer of lenalidomide, pomalidomide, avadomide, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, clathrate, or polymorph thereof.

127. The kit of any one of embodiments 103-126 wherein the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione, its stereoisomer or enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

128. The kit of any one of embodiments 103-127, wherein the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione.

129. The kit according to any one of embodiments 103-126, wherein the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione, its stereoisomer or enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

130. The kit of any of embodiments 103-127 and 129, wherein the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione .

131. The kit of any one of embodiments 103-130, wherein the immunomodulatory compound is formulated for oral, subcutaneous, or intravenous administration.

132. The kit of embodiment 131 wherein the immunomodulatory compound is formulated for oral administration.

133. The kit of any one of embodiments 103-132, wherein the immunomodulatory compound is formulated into capsules or tablets.

134. A set according to any one of embodiments 103-133, where:

each of one or more standard unit doses of an immunomodulatory compound contains an amount of from or about 0.1 mg to about 100 mg, from or about 0.1 mg to 50 mg, from or about 0.1 mg to 25 mg, from or about 0.1 mg to 10 mg, from or about 0.1 mg to 5 mg, from or about 0.1 mg to 1 mg, from or about 1 mg to 100 mg, from or about 1 mg to 50 mg, from or about 1 mg to 25 mg, from or about 1 mg to 10 mg, from or about 1 mg to 5 mg, from or about 5 mg to 100 mg, from or about 5 mg to 50 mg, from or about 5 mg to 25 mg, from or about 5 mg to 10 mg, from or about 10 mg to 100 mg, from or about 10 mg to 50 mg, from or from 10 mg up to 25 mg, from or about 25 mg to 100 mg, from or about 25 mg to 50 mg, or from or about 50 mg to 100 mg, each inclusive; and/or

each of one or more standard unit doses of an immunomodulatory compound contains at least, or about at least, 0.1 mg, 0.5 mg, 1.0 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg or 100 mg.

135. The kit of any one of embodiments 103-134, wherein each one or more standard unit doses of the immunomodulatory compound contain an amount greater than or greater than about 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg, and less than 25 mg.

136. The kit of any one of embodiments 103-135, wherein the T cell therapy is or comprises tumor infiltrating lymphocyte (TIL) therapy or engineered cells expressing a recombinant receptor that specifically binds to an antigen.

137. The kit of any one of embodiments 103-136, wherein the T cell therapy is or contains engineered cells expressing a recombinant receptor that specifically binds to an antigen.

138. The kit of embodiment 136 or embodiment 137, wherein the recombinant receptor is or contains a functional non-TCR antigen receptor or a TCR or antigen-binding fragment thereof.

139. The kit of any one of embodiments 136-138, wherein the recombinant antigen receptor is a chimeric antigen receptor (CAR).

140. The kit of any one of embodiments 136-139, wherein the recombinant antigen receptor comprises an extracellular domain containing an antigen-binding domain that specifically binds to an antigen.

141. The kit of any one of embodiments 136-140, wherein the antigen is associated with, specific to, and/or expressed on a cell or tissue of the disease, disorder, or condition.

142. The kit of embodiment 141 wherein the disease, disorder or condition is an infectious disease or disorder, an autoimmune disease, an inflammatory disease, or a tumor or cancer.

143. The kit of any one of embodiments 136-142, wherein the antigen is a tumor antigen.

144. The kit of any one of embodiments 136-143 wherein the antigen is selected from ROR1, B cell maturation antigen (BCMA), charcoal anhydrase 9 (CAIX), tEGFR, Her2/neu (erbB2 tyrosine kinase receptor), L1-CAM, CD19 , CD20, CD22, mesothelin, CEA and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40) , EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate-binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha , IL-13R-alpha2, kinase insert domain-containing receptor (kdr), kappa light chain, Ley antigen, L1 cell adhesion molecule, (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, preferably expressed melanoma antigen (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE Al, HLA-A2 NY-ESO-1, P SCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, testicular cancer antigen, mesothelin, mouse CMV, mucin 1 (MUC1 ), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, G protein coupled receptor 5D (GPCR5D), oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/ neu, estrogen receptor, progesterone receptor, aphrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms tumor 1 (WT-1), cyclin, cyclin A2, CCL-1, CD138 optionally, a human antigen to any of the above; a pathogen-specific antigen; and an antigen associated with a universal label.

145. The kit of any one of embodiments 136-144, wherein the antigen is or contains CD19, optionally human CD19.

146. The kit of any one of embodiments 136-145, wherein the antigen is or contains BCMA, optionally human BCMA.

147. The kit of any one of embodiments 136-146, wherein the antigen binding domain is or contains an antibody or antibody fragment, which is optionally a single chain fragment.

148. The kit of embodiment 147 wherein the fragment contains the variable regions of the antibody connected by a flexible linker.

149. The set of embodiment 147 or embodiment 148, wherein the fragment contains scFv.

150. The kit of any one of embodiments 136-149, wherein the recombinant receptor further comprises a spacer, optionally derived from an immunoglobulin, optionally containing a hinge region.

151. The kit of any one of embodiments 136-150, wherein the recombinant antigen receptor contains an intracellular signaling region.

152. The kit of embodiment 151 wherein the intracellular signaling region contains an intracellular signaling domain.

153. The kit of embodiment 152, wherein the intracellular signaling domain is or contains a primary signaling domain, a signaling domain that can induce a primary activation signal in a T-lymphocyte, a T-lymphocyte receptor (TCR) signaling domain component, and/or a signaling domain, containing an immunoreceptor tyrosine activating motif (ITAM).

154. The kit of embodiment 152 or embodiment 153, wherein the intracellular signaling domain is or comprises an intracellular signaling domain of the CD3 chain, optionally the CD3-zeta (CD3ζ) chain, or a signal portion thereof.

155. The kit of any one of embodiments 152-154, wherein the recombinant receptor further comprises a transmembrane domain located between an extracellular domain and an intracellular signaling region, where the transmembrane domain is optionally a CD8 or CD28 transmembrane domain.

156. The kit of any one of embodiments 152-155, wherein the intracellular signaling region further comprises a costimulatory signaling region.

157. The kit of embodiment 156, wherein the co-stimulatory signaling region comprises an intracellular signaling domain of a co-stimulatory T-lymphocyte molecule or a signaling portion thereof.

158. The kit of embodiment 156 or embodiment 157, wherein the costimulatory signaling region comprises a CD28, 4-1BB, or ICOS intracellular signaling domain, or a signal portion thereof.

159. The kit of any one of embodiments 156-158, wherein the co-stimulatory signaling region comprises the 4-1BB intracellular signaling domain.

160. The kit of any one of embodiments 156-159, wherein the costimulatory signaling region is located between the transmembrane domain and the intracellular signaling region.

161. The kit of any one of embodiments 103-160, wherein the T cell therapy comprises:

T lymphocytes selected from the group consisting of central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central memory T lymphocytes, effector T lymphocytes and regulatory T lymphocytes; and/or

a plurality of cells, this plurality contains at least 50% of a population of cells selected from the group consisting of CD4+ T lymphocytes, CD8+ T lymphocytes, central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central memory T-lymphocytes, effector T-lymphocytes and regulatory T-lymphocytes.

162. The kit of any one of embodiments 103-161, wherein the T cell therapy comprises T lymphocytes that are CD4+ or CD8+.

163. The kit of any one of embodiments 103-162, wherein the T cell therapy comprises primary cells obtained from a subject.

164. The kit of any one of embodiments 103-163, wherein the T cell therapy is autologous to the subject.

165. The method of any one of embodiments 103-164, wherein the T cell is allogeneic to the subject.

166. The kit of any one of embodiments 103-165, wherein the subject is a human.

167. The kit of any one of embodiments 103-166, wherein a single unit dose of T cell therapy contains from or about 1 x 10 ⁵ to 1 x 10 ⁸ cells, in total, T lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, from or about 5 × 10 ⁵ to 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total , or from or about 1 x 10 ⁶ to 1 x 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, each value inclusive.

168. The kit of any one of embodiments 103-167, wherein the single unit dose of T cell therapy comprises administering not more than 1 x 10 ⁸ cells, in total, T lymphocytes expressing recombinant receptors in total, or peripheral blood mononuclear cells (PBMCs). ), in total, not more than 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in general, or peripheral blood mononuclear cells (PBMC), in total, not more than 0.5 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, no more than 1 × 10 ⁶ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or mononuclear cells blood cells (PBMC), in general, no more than 0.5 × 10 ⁶ cells, in general, T-lymphocytes expressing recombinant receptors, in general, or peripheral blood mononuclear cells (PBMC), in general.

169. The kit of any one of embodiments 103-168, wherein the single unit dose of T cell therapy comprises a cell dose that is a divided dose, where the dose cells are administered in a plurality of compositions collectively containing all of the dose cells over a period of no more than three days.

170. The kit of any one of embodiments 103-169, wherein the instructions further specify administration of anti-lymphocyte chemotherapy prior to administration of T cell therapy.

171. The kit of any one of embodiments 103-170, wherein the disease or condition is a cancerous disease.

172. The kit of any one of embodiments 103-171, wherein the cancer is B-lymphocyte cancer and/or myeloma, lymphoma, or leukemia.

173. The kit of embodiment 171 or embodiment 172, wherein the cancer is mantle cell lymphoma (MCL), multiple myeloma (MM), acute lymphocytic leukemia (ALL), adult ALL, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma ( NHL) or diffuse large B-cell lymphoma (DLBCL).

174. The kit of embodiment 171 wherein the cancer is a non-hematologic cancer or is a solid tumor.

175. An article of manufacture comprising the kit of any one of embodiments 103-174.

176. A pharmaceutical composition containing T-cell therapy, an immunomodulatory compound and a pharmaceutically acceptable carrier.

177. The pharmaceutical composition of embodiment 176 wherein the T cell therapy is formulated in a single unit dose amount.

178. The pharmaceutical composition of embodiment 177, wherein a unit unit dose of T-cell therapy contains from or about 1 x 10 ⁵ to 1 x 10 ⁸ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or mononuclear cells of peripheral blood cells (PBMC), in total, from or about 5 × 10 ⁵ to 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, or from or from about 1 x 10 ⁶ to 1 x 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in general, or peripheral blood mononuclear cells (PBMC), in total, each value is inclusive.

179. The pharmaceutical composition of Embodiment 177 or Embodiment 178, wherein the single unit dose of T cell therapy comprises administering not more than 1 x 10 ⁸ cells, in total, of T lymphocytes expressing recombinant receptors in total, or peripheral blood mononuclear cells (PBMC), in total, not more than 1 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC), in total, not more than 0.5 × 10 ⁷ cells, in total, T-lymphocytes expressing recombinant receptors, in general, or peripheral blood mononuclear cells (PBMC), in total, not more than 1 × 10 ⁶ cells, in total, T-lymphocytes, expressing recombinant receptors, in total, or peripheral blood mononuclear cells (PBMC) in total not more than 0.5×10 ⁶ cells in total, recombinant receptor-expressing T-lymphocytes in total, or peripheral blood mononuclear cells (PBMC) in total.

180. The pharmaceutical composition of any one of embodiments 176-179, wherein the immunomodulatory compound binds to cereblon (CRBN) and/or the CRBN and E3 ubiquitin ligase complex and/or inhibits the transcription factor Ikaros (IKZF1) or Aiolos (IKZF3); and/or enhances ubiquitination or degradation of Ikaros (IKZF1) or Aiolos (IKZF3).

181. The pharmaceutical composition of any one of embodiments 176-180, wherein the immunomodulatory compound is thalidomide or is a derivative or analogue of thalidomide.

182. The pharmaceutical composition of any one of embodiments 176-181, wherein the immunomodulatory compound is lenalidomide, pomalidomide, avadomide, a stereoisomer of lenalidomide, pomalidomide, avadomide, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, clathrate, or polymorph thereof.

183. The pharmaceutical composition of any one of embodiments 176-182, wherein the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione , its stereoisomer or enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

184. The pharmaceutical composition of any one of embodiments 176-183, wherein the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione .

185. The pharmaceutical composition of any one of embodiments 176-182, wherein the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione, its stereoisomer or enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate or polymorph thereof.

186. The pharmaceutical composition of any one of embodiments 176-182 and 185, wherein the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6- dione.

187. The pharmaceutical composition of embodiment 176-186, wherein the immunomodulatory compound is formulated in a single unit dose amount.

188. Pharmaceutical composition according to any of the embodiments 176-187, where:

the amount of immunomodulatory compound in the composition is from or about 0.1 mg to about 100 mg, from or about 0.1 mg to 50 mg, from or about 0.1 mg to 25 mg, from or about 0.1 mg to 10 mg, from or about 0.1 mg to 5 mg, from or about 0.1 mg to 1 mg, from or about 1 mg to 100 mg, from or about 1 mg to 50 mg, from or about 1 mg to 25 mg, from or about 1 mg to 10 mg, from or about 1 mg to 5 mg, from or about 5 mg to 100 mg, from or about 5 mg to 50 mg, from or about 5 mg to 25 mg, from or about 5 mg to 10 mg, from or about 10 mg to 100 mg, from or about 10 mg to 50 mg, from or from 10 mg to 25 mg, from or from about 25 mg to 100 mg, from or from about 25 mg to 50 mg, or from or from about 50 mg to 100 mg, each inclusive; and/or

the amount of the immunomodulatory compound in the composition is at least, or about, at least 0.1 mg, 0.5 mg, 1.0 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg, or 100 mg.

189. The pharmaceutical composition of Embodiment 187 or Embodiment 188, wherein the amount of the immunomodulatory compound in the composition is greater than or about greater than 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg, and less than 25 mg.

190. The pharmaceutical composition of any one of embodiments 176-189, wherein the T cell therapy is or comprises tumor infiltrating lymphocyte (TIL) therapy or engineered cells expressing a recombinant receptor that specifically binds to an antigen.

191. The pharmaceutical composition of any one of embodiments 176-190, wherein the T cell therapy is or contains engineered cells expressing a recombinant receptor that specifically binds to an antigen.

192. The pharmaceutical composition of embodiment 190 or embodiment 191, wherein the recombinant receptor is or contains a functional non-TCR antigen receptor or a TCR or antigen-binding fragment thereof.

193. The pharmaceutical composition of any one of embodiments 190-192, wherein the recombinant antigen receptor is a chimeric antigen receptor (CAR).

194. The pharmaceutical composition of any one of embodiments 190-193, wherein the recombinant antigen receptor comprises an extracellular domain containing an antigen-binding domain that specifically binds to an antigen.

195. The pharmaceutical composition of any one of embodiments 190-194, wherein the antigen is associated with, specific to, and/or expressed on a cell or tissue of the disease, disorder, or condition.

196. The pharmaceutical composition of embodiment 195, wherein the disease, disorder, or condition is an infectious disease or disorder, an autoimmune disease, an inflammatory disease, or a tumor or cancer.

197. The pharmaceutical composition of any one of embodiments 190-195, wherein the antigen is a tumor antigen.

198. The pharmaceutical composition of any one of embodiments 190-197, wherein the antigen is selected from ROR1, B-cell maturation antigen (BCMA), charcoal anhydrase 9 (CAIX), tEGFR, Her2/neu (erbB2 tyrosine kinase receptor), L1-CAM, CD19, CD20, CD22, mesothelin, CEA and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40 ), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate-binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R- alpha, IL-13R-alpha2, kinase insert domain-containing receptor (kdr), kappa light chain, Ley antigen, L1 cell adhesion molecule, (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE-A3 , MAGE-A6, preferably expressed melanoma antigen (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE Al, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, testicular cancer antigen, mesothelin, mouse CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, G protein coupled receptor 5D (GPCR5D), oncofetal antigen, ROR1, TAG72, VEGF -R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms tumor 1 (WT-1) , cyclin, cyclin A2, CCL-1, CD138, optionally, a human antigen to any of the above; a pathogen-specific antigen; and an antigen associated with a universal label.

199. The pharmaceutical composition of any one of embodiments 190-198, wherein the antigen is or contains CD19, optionally human CD19.

200. The pharmaceutical composition of any one of embodiments 190-199, wherein the antigen is or contains BCMA, optionally human BCMA.

201. The pharmaceutical composition of any one of embodiments 190-200, wherein the antigen binding domain is or contains an antibody or antibody fragment, which is optionally a single chain fragment.

202. The pharmaceutical composition of embodiment 201, wherein the fragment contains antibody variable regions connected by a flexible linker.

203. The pharmaceutical composition of embodiment 201 or embodiment 202, wherein the fragment contains scFv.

204. The pharmaceutical composition of any one of embodiments 190-203, wherein the recombinant receptor further comprises a spacer, optionally derived from an immunoglobulin, optionally containing a hinge region.

205. The pharmaceutical composition of any one of embodiments 190-204, wherein the recombinant antigen receptor contains an intracellular signaling region.

206. The pharmaceutical composition of embodiment 205, wherein the intracellular signaling region contains an intracellular signaling domain.

207. The pharmaceutical composition of embodiment 206, wherein the intracellular signaling domain is or contains a primary signaling domain, a signaling domain that can induce a primary activation signal in a T-lymphocyte, a T-lymphocyte receptor (TCR) signaling domain component, and/or a signaling domain containing an immunoreceptor tyrosine activating motif (ITAM).

208. The pharmaceutical composition of embodiment 206 or embodiment 207, wherein the intracellular signaling domain is or contains an intracellular signaling domain of the CD3 chain, optionally the CD3-zeta (CD3ζ) chain, or a signal portion thereof.

209. The pharmaceutical composition of any one of embodiments 205-208, wherein the recombinant receptor further comprises a transmembrane domain located between an extracellular domain and an intracellular signaling region, where the transmembrane domain is optionally a CD8 or CD28 transmembrane domain.

210. The pharmaceutical composition of any one of embodiments 205-209, wherein the intracellular signaling region further comprises a costimulatory signaling region.

211. The pharmaceutical composition of embodiment 210, wherein the costimulatory signaling region comprises an intracellular signaling domain of a costimulatory T-lymphocyte molecule or a signaling portion thereof.

212. The pharmaceutical composition of embodiment 210 or embodiment 211, wherein the costimulatory signaling region comprises a CD28, 4-1BB, or ICOS intracellular signaling domain, or a signaling portion thereof.

213. The pharmaceutical composition of any one of embodiments 210-212, wherein the co-stimulatory signaling region comprises the 4-1BB intracellular signaling domain.

214. The pharmaceutical composition of any one of embodiments 210-213, wherein the costimulatory signaling region is located between the transmembrane domain and the intracellular signaling region.

215. The pharmaceutical composition of any one of embodiments 210-214, wherein the recombinant receptor is or contains a chimeric antigen receptor containing an antigen-binding domain, a spacer, a transmembrane domain from CD28, an intracellular signaling domain containing the CD3-zeta chain (CD3ζ) and an intracellular signaling domain from 4-1BB.

216. The pharmaceutical composition of any one of embodiments 176-215, wherein the T cell therapy comprises:

T lymphocytes selected from the group consisting of central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central memory T lymphocytes, effector T lymphocytes and regulatory T lymphocytes; and/or

a plurality of cells, this plurality contains at least 50% of a population of cells selected from the group consisting of CD4+ T lymphocytes, CD8+ T lymphocytes, central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central memory T-lymphocytes, effector T-lymphocytes and regulatory T-lymphocytes.

217. The pharmaceutical composition of any one of embodiments 176-216, wherein the T cell therapy comprises T lymphocytes that are CD4+ or CD8+.

218. The pharmaceutical composition of embodiment 217 wherein the ratio of CD4+ T lymphocytes to CD8+ T lymphocytes is from or about 1:3 to 3:1, optionally 1:1.

219. The pharmaceutical composition of any one of embodiments 176-218, wherein the T cell therapy comprises primary cells obtained from a subject.

220. The pharmaceutical composition of embodiment 219, wherein the subject is a human.

221. A pharmaceutical composition according to any one of embodiments 176-220, containing a volume of from or about 1 ml to 100 ml, 1 ml to 75 ml, 1 ml to 50 ml, 1 ml to 25 ml, 1 ml to 10 ml, 1 ml to 5 ml, 5 ml to 100 washes 5 ml to 75 ml, 5 ml to 50 ml, 5 ml to 25 ml, 5 ml to 10 ml, 10 ml to 100 ml, 10 ml to 75 ml, 10 ml to 50 ml, 10 ml to 25 ml, 25 ml to 100 washes 25 ml to 75 washes 25 ml to 50 ml, 50 ml to 100 ml, 50 ml to 75 ml or from 75 ml to 100 ml.

222. A pharmaceutical composition according to any one of embodiments 176-221, containing a volume of at least, or about, at least, or about 1 ml, 5 ml, 10 ml, 20 ml, 25 ml, 30 ml, 40 ml, 50 ml, 60 ml, 70 ml, 80 ml, 90 ml or 100 ml.

223. The pharmaceutical composition of any one of embodiments 176-222 further comprising a cryoprotectant.

224. The pharmaceutical composition of any one of embodiments 176-223, which is sterile.

225. An industrial product containing a pharmaceutical composition according to any of the embodiments of 176-223.

226. A method of treatment comprising administering a pharmaceutical composition according to any one of embodiments 176-225 to a subject for the treatment of a disease or condition.

227. The method of embodiment 226 wherein the disease or condition is cancer.

228. The method of embodiment 227 wherein the cancer is B-lymphocyte cancer and/or myeloma, lymphoma, or leukemia.

229. The method of embodiment 216 or embodiment 228 wherein the cancer is mantle cell lymphoma (MCL), multiple myeloma (MM), acute lymphocytic leukemia (ALL), adult ALL, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL ) or diffuse large B-cell lymphoma (DLBCL).

230. The method of embodiment 227 wherein the cancer is a non-hematologic cancer or is a solid tumor.

VII. Examples

[0633] The following examples are included for illustrative purposes only and are not intended to limit the scope of the present invention.

Example 1 Anti-BCMA CAR T cell cytolytic activity and cytokine production after incubation with target cell lines expressing BCMA in the presence or absence of lenalidomide

[0634] T-lymphocytes are isolated by immunoaffinity-based enrichment from leukapheresis samples from healthy donors. The isolated cells are transduced with a viral vector encoding one of a variety of exemplary anti-BCMA CARs. Each anti-BCMA CAR contains a human anti-BCMA scFv, a spacer region, a CD28 transmembrane domain, a 4-1BB-derived intracellular co-signaling sequence, and an intracellular signaling domain derived from CD3-zeta. The viral vector construct further encodes a truncated EGFR (EGFRt) that serves as a surrogate marker for CAR expression; The EGFRt-coding region is separated from the CAR sequence by the T2A hopping sequence. After transduction, cells proliferate and the resulting compositions are frozen by cryopreservation.

[0635] Cryo-frozen anti-BCMA CAR+ T lymphocytes are thawed and evaluated for various responses after co-culture with BCMA-expressing target cells in the presence or absence of lenalidomide. In vitro assays to assess destruction target cells and cytokine production are performed using two different target multiple myeloma cell lines expressing BCMA RPMI-8226 or OPM-2. FIG. 1A shows surface BCMA expression assessed by flow cytometry after anti-BCMA antibody staining of exemplary multiple myeloma cell lines including RPMI-8226 and OPM-2. The dashed line shows the background of a BCMA-negative cell line stained with anti-BCMA antibody. MFI is the median fluorescence intensity. BCMA expression is relatively low for both cell lines (see Lee et al. (2016) Br J Haematol. 174:911-922). RPMI-8226 has been shown to be more sensitive to lenalidomide compared to OPM-2 (6.43 and 37.4 µM, respectively) (Wellcome Sanger Institute. Genomics of drug sensitivity in cancer. www.cancerrxgene.org/translation/ Drug/1020, Accessed February 7, 2018).

A. RPMI-8226

1. Cytolytic activity

[0636] Cells of a target cell line expressing BCMA (RPMI-8226) are incubated with exemplary anti-BCMA CAR T lymphocytes expressing human scFv anti-BCMA CARs at an effector to target cell ratio (E:T) of 0, 3:1 in the presence of 1 μM or 10 μM lenalidomide or without lenalidomide (carrier medium). Co-cultures with T lymphocytes not expressing CAR (sham) or cultures with only target cells (no CAR T) are used as controls, each with or without (carrier medium) 10 μM or 1 μM lenalidomide. Cells from each condition are inoculated in triplicate.

[0637] Target RPMI-8226 cells are labeled with NucLight Red (NLR) to enable them to be tracked by microscopy. Cytolytic activity is assessed by measuring the loss of viable target cells over a period of six days, as determined by the red fluorescence signal (using the IncuCyte® Live Cell Analysis System, Essen Bioscience). Normalized target cell counts are generated by dividing the target cell counts by the cell counts at the start of each culture. Percent target cell death is assessed by measuring the area under the curve (AUC) for normalized target cell counts as a function of time and normalizing reciprocal AUC values (1/AUC) by determining a value of 0% (target cells only) and a value of 100% (CAR+ T cells). co-cultured with target cells in vehicle control).

[0638] As shown, co-cultivation in the presence of 1 μM (FIG. 1C) or 10 μM (FIGS. 1B, 1C) lenalidomide results in increased target cell death by CAR+ anti-BCMA T-lymphocytes on day 6 of co-cultivation, by compared to incubation of the target cells together with anti-BCMA CAR+ T lymphocytes in the absence of lenalidomide (corresponding to 100% in FIG. 1B). As shown in FIG. 1C, the observed effect of lenalidomide on cytolytic activity is dose-sensitive and delayed, not appearing until approximately 50 hours in culture. The results are consistent with the role of lenalidomide in facilitating continued function and/or survival (eg, by preventing cell depletion or cell death) of CAR T lymphocytes after initial activation. Similar results are observed for engineered cells expressing a number of other anti-BCMA CARs, each containing different scFv binding domains.

2. Production/accumulation of cytokines

[0639] The levels of various cytokines are assessed in culture supernatants after incubation of anti-BCMA CAR+ T-lymphocytes with cells of the target cell line expressing BCMA, RPMI-8226 at an effector to target cell ratio (E:T) of 0.3:1 in the presence of or in the absence of 10 μM lenalidomide. A culture of T-lymphocytes not expressing CAR anti-BCMA (sham) was used as a control. The amounts of IL-2 (FIG. 2A), IFNγ (FIG. 2B) and TNF-α (FIG. 2C) in culture supernatants were assessed 48 hours after the start of culture. As shown in Figures 2A-2C, the presence of lenalidomide is associated with an increase in CAR-dependent cytokine production and/or accumulation after co-culture of anti-BCMA CAR+ T-lymphocyte target cells with antigen-specific target cells. These results are consistent with the role of lenalidomide in facilitating CAR-mediated effector functions. Similar results are observed for cells expressing various other anti-BCMA CARs each containing different scFv binding domains.

B.OPM-2

3. Cytolytic activity

[0640] OPM-2 multiple myeloma target cells are incubated with human T lymphocytes (isolated from four different independent donors) expressing exemplary anti-BCMA CAR) at a 1:1 ratio of effector to target cells (E:T) in the presence of 0.01 µM, 0.1 µM, 1.0 µM or 10 µM lenalidomide or in the absence of lenalidomide, over a period of 7 days. OPM-2 cells are labeled with NucLight Red (NLR) to enable tracking of target cells by microscopy, essentially as described above. Cytolytic activity is assessed by measuring the loss of viability of target cells at the end of incubation. The degree of cytolytic activity observed for cultures incubated in the absence of lenalidomide is set as background, 100%. The results are shown in Fig. 3A. The addition of lenalidomide was observed to improve the cytolytic activity of anti-BCMA CAR+ T-lymphocytes against target OPM-2 cells in a dose-dependent manner. Similar results are observed for other anti-BCMA CAR-expressing T-lymphocytes, including cells expressing different anti-BCMA CARs each containing different scFv binding domains and/or engineered cells using cells from different donors.

4. Cytokine

[0641] Anti-BCMA CAR+ T lymphocytes from four independent donors are incubated with a target cell line expressing BCMA, OPM-2 at an effector to target cell ratio (E:T) of 1:1 in the presence of 0.01 μM, 0.1 μM, 1.0 μM or 10 μM lenalidomide or in the absence of lenalidomide (background, corresponds to 100%). After 24 hours of culture, the presence of IFNγ (FIG. 3B), IL-2 (FIG. 3C) and TNF-α (FIG. 3D) in the culture supernatants is assessed. As shown in Figures 3B-3D, lenalidomide was observed to increase cytokine production and/or accumulation by antigen-stimulated CAR+ anti-BCMA T lymphocytes in a dose dependent manner.

C. Comparison of multi-donor anti-BCMA activity of CAR+ T-lymphocytes

[0642] In another study, anti-BCMA CAR T lymphocytes from a representative healthy donor and a multiple myeloma patient (the patient was pomalidomide refractory) are incubated with fluorescently labeled OPM-2 target cells at an effector to target cell ratio (E: T) 0.3:1 in the presence of various concentrations of lenalidomide (0.01 μM, 0.1 μM, 1.0 μM or 10 μM lenalidomide) or in the absence of lenalidomide, for 6-7 days. Cytolytic activity is measured by the loss of red fluorescent cells. To assess cytokine production, anti-BCMA CAR T lymphocytes obtained from a healthy donor and from a multiple myeloma patient are co-cultured with fluorescently labeled OPM-2 target cells at an effector to target cell ratio (E:T) of 1:1 in the presence of various lenalidomide concentrations (0.01 μM, 0.1 μM, 1.0 μM or 10 μM lenalidomide) or in the absence of lenalidomide. After 24 hours, take samples of media to assess the presence of IFNγ and IL-2. The results are shown in Fig.3E, they represent the average of two experiments; antigen-specific cytolytic activity of CAR-T anti-BCMA and cytokine production have been observed to be increased by lenalidomide in a concentration dependent manner.

[0643] The studies discussed above extend to anti-BCMA CAR+ T lymphocytes generated from cells from two additional healthy donors. Anti-BCMA CAR+ T-lymphocyte activities from three healthy donors and one IMiD refractory patient (donor patient was refractory to pomalidomide) were compared to multiple myeloma cell lines expressing BCMA, both OPM-2 and RPMI-8226. Cytolytic activity and production of cytokines (IFNγ, IL-2 and TNF-α) are analyzed essentially as described above. The absolute changes in cytokine levels relative to the control carrier medium are calculated. Experiments are carried out 2-3 times on each donor.

[0644] Increased cytolytic activity of CAR T anti-BCMA against OPM-2 target cells titrated with increasing concentrations of lenalidomide is observed in all donors (P=6.2×10 ^-5 ) (FIG. 3F). As shown in FIG. 3F, the effect of lenalidomide treatment on CAR T cytolytic activity in the RPMI-8226 coculture appears to be donor dependent, with the donor patient showing a significant increase in cytolytic activity (P=1.9 x 10 ^-8 ). In addition, all CAR T donors have significantly increased production of IFN-γ, IL-2 and TNF-α in a lenalidomide concentration-dependent manner in co-culture with OPM-2 cells (P<0.002, Fig. 3G). Cytokine production by CAR-expressing T lymphocytes co-cultured with RPMI-8226 was also significantly increased for all donors and cytokines by lenalidomide treatment (P<0.003, Fig. 3H).

Example 2 Effects of Lenalidomide on CAR T-Lymphocyte Proliferation and Antigen-Specific Function in Serial Restimulation

A. Proliferation of CAR T-lymphocytes

[0645] The ability of CAR T lymphocytes to proliferate and exhibit antigen-specific function ex vivo after repeated rounds of antigen stimulation may correlate with in vivo function and/or the ability of cells to persist in vivo (e.g., after administration and initial activation in response to encounter with antigen) (Zhao et al. (2015) Cancer Cell, 28:415-28). Anti-BCMA CAR+ T lymphocytes generated as described above are inoculated in triplicate at 1 x 10 ⁵ cells/well in 96-well plates. Irradiated target cells expressing BCMA (MM1.S cells) are added at an effector to target cell ratio (E:T) of 1:2 in the presence or absence of various concentrations (0.01 μM, 0.1 μM, 1.0 μM or 10 μM) lenalidomide.

[0646] Every 3-4 days (at the start of each new entry), CAR T-lymphocytes are counted. Cells are then harvested and re-inoculated at initial seeding density with fresh media, newly added lenalidomide at the same concentration, where applicable, and re-thawed, re-irradiated target cells. Carry out 8 visits stimulation during the cultivation period of 31 days. For some visits, upon re-inoculation, cells are assessed for phenotypic markers using flow cytometry.

[0647] Illustrative results are shown in Fig. 4A. As shown, an increase in anti-BCMA CAR T lymphocyte proliferation was observed at day 14 at all lenalidomide concentrations compared to wells without lenalidomide. The assay is performed on different compositions of anti-BCMA CAR+ T lymphocytes, each generated by injecting CAR into T lymphocytes obtained from one of six different donors. The assay is performed on genetically engineered cells from six different independent donors for CAR expression. For each donor, an increase or no change in CAR-T expansion was observed at a concentration of 0.1 μM lenalidomide. 4B shows the results of a similar assay in which genetically engineered cells to express two different human anti-BCMA CARs are exposed to multiple rounds of target cell stimulation in the presence or absence of lenalidomide. As shown, the presence of lenalidomide in cultures was observed to cause an increase in proliferation in both cell populations beginning between day 21 and day 28. Results are consistent with the conclusion that lenalidomide may facilitate continued expansion of CAR+ T-lymphocytes and/or survival after re-encountering a recognized antigen.

B. CAR T-lymphocyte counts, cytokine production and activation

[0648] 3-donor CAR+ anti-BCMA T lymphocytes generated as described above are inoculated in triplicate on 96-well plates with irradiated target cells expressing BCMA (MM1S cells) at an effector to target cell ratio (E:T ) 1:2 in the presence of 0.1 μM lenalidomide or vehicle control. Culture conditions are set anew every 3-4 days. Reinoculation is maintained for 28 days or until the cell count is <50,000 cells. The experiments are carried out in triplicate on 3 donors. Cytokine levels (IFNγ, IL-2, and TNF-α) are assessed 24 hours after reinoculation on days 5, 8, and 15. CAR T-lymphocyte activation is measured on cells harvested on days 4, 7, and 14 by flow cytometry for CD25.

[0649] FIG. 5A shows the cell counts (doubling of the projective population) of anti-BCMA CAR+ T lymphocytes for each restimulation time point. “×” indicates insufficient number of cells to reinoculate in the assay. The results show that after re-stimulation with target cells, all 3 lenalidomide-treated CAR T donors have increased projective cell counts at 28 days compared to controls (P < 0.003). FIG. 5B shows median CD25 fluorescence intensity (MFI) (tuned for live CAR+ CD3+) and FIG. 5C shows cytokine production normalized to number of cells inoculated. The increase in cell counts is associated with a significant increase in CD25 expression in CAR T (P<3.4×10 ^-4 ; Fig.5B) and the production of IL-2, IFN-γ and TNF-α in media (P<0.5; Fig. .5C). The results show that anti-BCMA CAR T-lymphocyte counts, cytokine production and activation are increased by lenalidomide after in vitro re-stimulation.

Example 3 Effects of Lenalidomide on Proliferation and Activation of CAR-T BCMA in a 3D Myeloma Model

[0650] To evaluate cell function in the context of the three-dimensional (3-D) microenvironment of tissue expressing human BCMA, reshaped bone marrow (rBone™) (zPREDICTA, San Jose, CA) is immersed in BCMA-expressing RPMI-8226. 20,000 T lymphocytes expressing another exemplary human anti-BCMA CAR (or mock T lymphocytes not expressing CAR) are incubated in a 3-D model in the presence or absence of 1.0 μM lenalidomide.

[0651] After 2 or 7 days, cells are isolated and evaluated by flow cytometry for surface expression of CD3, CD25, CD4 and CD8. As shown in Figure 6A, the presence of lenalidomide was observed to result in an increase in total CD3+ cell counts in anti-BCMA CAR+ T cell cultures on day 7. An increase in CD25+ expression in CD4+ T cell populations (Figure 6B) and CD8+ (Fig.6C) is also observed in the presence of lenalidomide. The results are consistent with the conclusion that lenalidomide can facilitate the increase in multiplication, survival and/or function of anti-BCMA CAR+ T lymphocytes in the antigen-expressing tumor microenvironment.

Example 4 Effect of lenalidomide on CAR T-lymphocyte function in vivo

[0652] The anti-tumor effects of CAR+ anti-BCMA T-lymphocytes alone and in combination with lenalidomide were evaluated in two different BCMA-expressing mouse tumor models, the grafted human multiple myeloma model RPMI 8226 (subcutaneous implant model) and the OPM multiple myeloma model -2 human grafted mice (orthotropic bone marrow model).

A. Model RPMI-8226

[0653] Mice are injected subcutaneously (sc) with 5 x 10 ⁶ RPMI-8226 cells and allowed to grow tumor volume to approximately 150 mm ³ . On day 0, a composition containing a suboptimal (low) dose of anti-BCMA CAR+ T-lymphocytes (generated by transducing cells derived from samples of human donor subjects, essentially as described above) is administered intravenously to mice (together with a similar composition of T- non-CAR-expressing lymphocytes (sham) used as controls). Specifically, the composition contains approximately 5×10 ⁵ CAR+ (or sham) CD4+ T lymphocytes and 5×10 ⁵ CAR+ (or sham) CD8+ T lymphocytes. T-lymphocytes are adoptively transferred into mice, alone or in combination with lenalidomide given daily at 25 mg/kg, intraperitoneally (ip) starting on day 0 (along with T-lymphocyte administration) continuing until day 21. B the other control group of mice is injected with lenalidomide itself, without the introduction of T-lymphocytes. Tumor volume and animal survival are monitored throughout the study. Blood from the retro-orbital (RO) circulation was collected weekly for assessment of plasma levels of BCMA and IFN-gamma and for pharmacokinetic (PK) assessment of CAR+ T-lymphocytes.

[0654] Tumor volume measurements are shown for individual animals in FIG. 7A, administration of lenalidomide and a low dose of anti-BCMA CAR+ T lymphocytes in combination was observed to result in slower tumor growth compared to mice treated with lenalidomide or T alone. -lymphocytes CAR+ anti-BCMA. The observed effect is most evident at later time points, including times after the last daily administration of lenalidomide (ie, after day 21). The results are consistent with the ability of lenalidomide to increase the capacity of T-lymphocytes for long-term persistence and/or function.

[0655] As shown in Fig.7B, mice that received lenalidomide and anti-BCMA CAR+ T-lymphocytes show an increase in survival compared to other treatment groups. The median survival (ms) of mice treated with CAR+ anti-BCMA T-lymphocytes and lenalidomide is 85 days (twice as long as the other treatment groups, which show a median survival of 38-43.5 days).

[0656] In addition, the number of CAR+ CD4+ and CD8+ T-lymphocytes and T-lymphocytes without CAR in the peripheral blood of each animal is determined on days 7, 14, 21 and 34. CAR CD4+ T-lymphocytes and T-lymphocytes without CAR are shown in Figures 8A and 8E (days 7 and 14), respectively, and in Figures 8B and 8F (days 21 and 34), respectively. The numbers of CAR CD8+ T lymphocytes and non-CAR T lymphocytes are shown in Figures 8C and 8G (days 7 and 14), respectively, and in Figures 8D and 8H (days 21 and 34), respectively. As shown, an increase in the number of CAR+ CD4+ and CD8+ T lymphocytes (but not non-CAR+ T lymphocytes) in the blood was observed on day 36 in mice receiving a combination of anti-BCMA CAR+ T lymphocytes and lenalidomide, compared with other treatment groups.

B. Model Opm-2

i. Study 1

[0657] The effect of lenalidomide in combination with CAR T anti-BCMA is also evaluated in an orthotopic mouse tumor model using OPM-2 cells. Mice (NOD.Cg-Prkdc ^scid IL-2rg ^tm1Wjl /SzJ mice (NSG; Jackson Labs)) are intravenously injected with (iv) 2 x 10 ⁶ OPM2 (multiple myeloma) cells transfected with firefly luciferase (OPM2-ffluc). The tumor is allowed to engraft for 13 days before staging (14 days before CAR T-lymphocyte injection) and engraftment is verified using bioluminescent imaging. Mice are administered one or more compositions in different treatment groups, as follows and is given in Table E1.

[0658] Some groups receive 10 mg/kg of lenalidomide in phosphate-buffered saline via intraperitoneal injection, either (A) starting on day -1 (the day before administration of CAR+ T-lymphocytes) (lenalidomide (A)); or (B) on day 14 (day 14 after the start of CAR+ T-lymphocyte administration) (lenalidomide (B)), in each case, daily, for the duration of the study. In CAR+ T-lymphocyte receiving groups, CAR anti-BCMA (generated by transducing cells obtained from samples from human donor subjects, essentially as described above) is administered on day 0 (day 14 after tumor cell injection), at a dose of either 5 x 10 ⁵ (low) or 1 x 10 ⁶ (high) CAR-expressing T-lymphocytes. Table E1 lists dosing regimens.

Table E1: Study Design Group No. Group Description Injected CAR T-lymphocytes (or sham-transduced
T-lymphocytes) one Only tumor 0 2 simulation (high) (1 × 10 ⁶ ) 3 Lenalidomide (A) No four Lenalidomide (B) No 5 Imitation+lenalidomide (A) (1 × 10 ⁶ ) 6 Imitation+lenalidomide (B) (1 × 10 ⁶ ) 7 T-lymphocytes CAR+ anti-BCMA (high) 1 x ¹⁰⁶ eight T-lymphocytes CAR+ anti-BCMA (low) ⁵ ×105 9 T-lymphocytes CAR+ anti-BCMA (high) + lenalidomide (A) 1 x ¹⁰⁶ ten T cells CAR+ anti-BCMA (low) + lenalidomide (A) ⁵ ×105 eleven T-lymphocytes CAR+ anti-BCMA (high) + lenalidomide (B) 1 x ¹⁰⁶ 12 T cells CAR+ anti-BCMA (low) + lenalidomide (B) ⁵ ×105

[0659] The mass of the tumor in animals among in different groups tracked using bioluminescent images until day 39 after dosing T-lymphocytes CAR+. For bioluminescent imaging, mice receive intraperitoneal (i.p.) injections of luciferin substrate (CaliperLife Sciences, Hopkinton, MA) resuspended in PBS (15 μg/g body weight). The total luminous flux (photons/sec) is determined at each moment of time.

[0660] Figures 9A and Figure 9B show results for tumor weight up to day 46, in mice treated with lenalidomide daily starting on day -1 (lenalidomide A), in the presence or absence of high (1×10 ⁶ ; Fig. 9A) or low (5 x 10 ⁵ ; Fig. 9B) dose of CAR+ T-lymphocytes. Figure 9C shows graphs for tumor mass in individual animals up to day 53. Figure 9D shows graphs and tumor imaging results (day 46 after CAR+ cell injection) for individual animals receiving a higher dose of CAR+ with lenalidomide on day -1 (lenalidomide A ). 9E shows graphs and tumor imaging results (day 46 after CAR+ cell injection) for selected animals receiving a higher dose of CAR+ without lenalidomide on day -1 (lenalidomide A). The asterisks indicate the death or killing of an individual animal at the time shown in the graph. As shown, the addition of lenalidomide has been observed to result in slower tumor growth and reduced tumor mass in mice administered with CAR+ T-lymphocytes at both doses of CAR+ T-lymphocytes.

[0661] Fig.9F and Fig.9G show the results of tumor weight at time point, in studies on mice administered lenalidomide, starting from day 14 after the introduction of T-lymphocytes CAR+ (lenalidomide B) (vertical lines in Figures 9F and 9G ) in the presence or absence of a high (1 x 10 ⁶ , Fig. 9F) or low (5 x 10 ⁵ , Fig. 9G) dose of CAR+ T-lymphocytes. Figure 9H shows plots for tumor weight of individual animals up to day 53. Figure 9I shows plots and tumor imaging results (day 46 after CAR+ cell injection) for individual animals receiving a higher dose of CAR+ with lenalidomide on day -1 (lenalidomide A) . 9J shows graphs and tumor imaging results (day 46 after CAR+ cell injection) for selected animals receiving a higher dose of CAR+ without lenalidomide on day -1 (lenalidomide A). As shown, while lenalidomide alone was not observed to reduce tumor growth or tumor mass, the addition of lenalidomide was observed to result in a trend towards slowing tumor growth and decreasing tumor mass in mice treated with both doses of T -lymphocytes CAR, with clear differences observed starting on day 30-40 after the injection of T-lymphocytes CAR, for a higher (1 × 10 ⁶ ) dose of T-lymphocytes CAR+. At this dose, combination with lenalidomide has been observed to slow tumor growth, whether given on day -1 or via delayed dosing.

[0662] The results of survival at time point in the studies are shown in Figures 10A and 10B (for groups receiving lenalidomide with regimens (d-1) and B (d.14 after CAR (delayed) respectively), and Figures 10C and 10D ( for high and low dose CAR groups, respectively.) The addition of lenalidomide has been shown to improve the impact on survival observed in mice treated with anti-BCMA CAR+ T-lymphocytes (at both high and low doses evaluated) when they administered either on day -1 (A) or via delayed (d.14) dosing (B).

[0663] Table E2 lists the median survival (ms) (as assessed at day 56 post-treatment with CAR+ T-lymphocytes) and the number of mice per group surviving to day 56 post-treatment with CAR+ T-lymphocytes for each group of animals evaluated in this study .

TABLE E2: SURVIVAL Group Dose of cells Median survival
(assessed on day 6) Number of animals surviving day 56 after CAR T simulation (high) 1.00E+06 24 0/8 Lenalidomide (A) N/A 28 0/8 Lenalidomide (B) N/A 25 0/8 Imitation+lenalidomide (A) 1.00E+06 28 0/8 Imitation+lenalidomide (B) 1.00E+06 25 0/8 T-lymphocytes CAR+ (high) 1.00E+06 56 2/8 T-lymphocytes CAR+ (low) 5.00E+05 35 0/8 T-lymphocytes CAR+ (high) + lenalidomide (A) 1.00E+06 N/A 6/8 T-lymphocytes CAR+ (low) + lenalidomide (A) 5.00E+05 52 1/8 T-lymphocytes CAR+ (high) + lenalidomide (B) 1.00E+06 N/A 6/8 T cells CAR+ (low) + lenalidomide (B) 5.00E+05 36 2/8

(ii.) Study 2

[0664] In additional studies, NOD/Scid/gc ^-/- (NSG) mice are injected with (iv) OPM-2-luciferase cells as described in Study 1 above and allowed to engraft for 14 days prior to infusion (iv) CAR-T (or sham) cells. In some groups, daily intraperitoneal administration of 10 mg/kg of lenalidomide or vehicle control is started either on day -1 (the day before CAR-T administration) (simultaneously with lenalidomide (lenalidomide (C) or vehicle (vehicle (C)), or on day 14 after administration of CAR (or sham) T-lymphocytes (delayed lenalidomide (D)).

[0665] 14 days after tumor cell injection (day 0), an intravenous injection of a sub-therapeutic dose of CAR+ T lymphocytes (1 x 10 ⁶ CAR T lymphocytes (generated from two different donors)) or sham control cells is given. The results are shown in Fig.10E, 10F, 10G and 10H. Data are presented as mean ± SEM (standard deviation). For in vivo survival, the Gehan-Breslow-Wilcoxon test is used to compare groups.

[0666] FIG. 10E shows the results of tumor weight assessment up to day 60 as analyzed by bioluminescence measured by flow cytometry. The addition of lenalidomide has been observed to result in slower tumor growth and reduced tumor mass in mice treated with CAR+ T-lymphocytes generated from both donor cells. As shown in FIG. 10F, the addition of lenalidomide has been observed to improve the impact on survival in mice treated with anti-BCMA CAR+ T-lymphocytes, especially after co-administration of lenalidomide. Fixed-exposure or mixed-exposure linear models are used to assess the significance of lenalidomide treatment for cytolytic activity, with treatment, donor, and time considered as fixed exposures, and the animal treated as random exposure nested over time, with repeated measurements obtained from one and the same time. the same animal. P values are obtained by probability ratio tests comparing the full model with the impact of interest to the model without the impact of interest. Simultaneous addition of lenalidomide leads to a significant decrease in tumor mass for donor 1 (P=0.02) and an increase in survival for donor 1 (P=0.057) and donor 2 (P=0.04), compared with vehicle treated animals, who are injected with CAR T anti-BCMA only. Animals on the simultaneous dosing regimen of lenalidomide also show an increase in peripheral blood CAR T counts after 7 days (P=7.3×10 ^-6 ), but at later time points. Lenalidomide had a small but significant effect, along with sham CAR T, on tumor mass for donor 1 only (P=0.003). In this study, the addition of delayed dosing lenalidomide did not improve tumor vanishing and survival for both CAR T donors. The results show that survival and tumor vanishing with a subtherapeutic dose of CAR-T anti-BCMA is enhanced by lenalidomide in an in vivo OPM-2 tumor model.

[0667] Blood from treated mice is collected for CAR-T pharmacokinetic analysis and cells are stained with antibodies to exclude mouse specific cells (H2-kd, TER119 and muCD45) and analyzed by flow cytometry. Cells are gated to CAR+ CD45+ and CD3+ and the number of cells per microliter of blood is determined. For pharmacokinetic measurements, each time point is analyzed using a one-way ANOVA test and Tukey's post-hoc test. Figures 10G and 10H show flow cytometric analysis of sham control cells and CAR T lymphocytes in the blood of mice on days 8, 14, 22 and 28 after injection of CAR T lymphocytes from two donors. The results show that there is an increase in peripheral blood CAR T-lymphocyte counts at early time points, in particular after co-administration of lenalidomide (**P<0.01).

Example 5 Effects of lenalidomide on anti-CD19 CAR proliferation upon suboptimal stimulation

[0668] Anti-CD19 CAR-expressing T lymphocytes are generated by genetically engineering CD4+ and CD8+ T lymphocytes (which are isolated by immunoaffinity-based enrichment in healthy human donor subjects) with a viral vector encoding anti-CD19 CAR. CAR containing anti-CD19 scFv, an Ig-derived spacer, a transmembrane domain derived from human CD28, an intracellular signaling domain derived from human 4-1BB, and a signaling domain derived from human CD3 zeta. The nucleic acid construct encoding CAR also contains a truncated EGFR sequence (tEGFR) for use as a transduction marker, separated from the CAR sequence by a T2A self-cleaving sequence.

[0669] Anti-CD19 CAR T lymphocytes are suboptimally stimulated by incubation with anti-CD3 (without a second reagent such as anti-CD28 engineered to produce a costimulatory signal), in the presence of 5 μM lenalidomide or vehicle control. CAR-expressing anti-CD19 T lymphocytes are labeled with CELLTRACE VIOLET (CTV; ThermoFisher Scientific, Waltham MA) prior to incubation; proliferation is assessed by assessing the dilution of the dye using flow cytometry. As shown in FIG. 11, in the context of suboptimal stimulation conditions over a period of 72 hours, lenalidomide was observed to have increased proliferation of CAR+ T lymphocytes.

Example 6 Observed relationship between treatment outcomes and peripheral blood CAR+ T-lymphocyte levels in a cohort of human subjects administered CAR-expressing anti-CD19 cells.

[0670] Treatment outcomes and blood CAR+ T-lymphocyte counts are evaluated in twenty-eight adult subjects with relapsing or refractory (R/R) non-Hodgkin's lymphoma (NHL) treated with autologous T-lymphocytes expressing a CD19-targeted chimeric antigen receptor (CAR) containing an anti-CD19 scFv antibody and a 4-1BB intracellular signaling domain (administered at a ratio of CD4+ to CAR+ CD8+ T-lymphocytes of approximately 1:1).

[0671] Prior to administration of CAR-expressing T-lymphocytes, subjects are treated with 30 mg/m ² fludarabine daily for 3 days and 300 mg/m ² cyclophosphamide daily for 3 days. At d=0, subjects are treated with 5 x 10 ⁷ (DL-1) or 1 x 10 ⁸ (DL-2) CAR-expressing T lymphocytes by intravenous infusion.

[0672] The response rates observed at a particular time point in the ongoing study are shown in Table E3 for a cohort of 20 subjects with diffuse large B-cell lymphoma (DLBCL) treated with a single dose of DL-1. As shown, an overall response rate (ORR) of 80% (16/20) is observed, and 60% (12/20) of subjects are observed to achieve complete remission (CR). 20% (4/20) of subjects show a partial response (PR) and 20% (4/20) show progressive disease (PD). Of chemotherapy-refractory subjects (showing stable or progressive disease after the last chemotherapy regimen or disease recurrence less than 12 months after autologous SCTs) before administration of CAR+ T-lymphocytes, the overall response rate is 83% (10 ORR, 7 CR, 3 PR, 2PD, n=12). Among non-responders (showing less than complete remission after the last treatment but not considered non-responders), the overall response rate is 77% (13 ORR, 9 CR, 4 PR, 4PD, n=17).

Table E3. General reaction DLBCL cohort, DL1 single dose regimen All (n=20) Not amenable
treatment ^*
(n=17)

(n=12)Not amenable

(n=12) ORR, n (%) [95% CI] 16 (80) [56, 94] 13 (77) [50, 93] 10 (83) [52, 98] CR, n (%) [95% CI] 12 (60) [36, 81] 9 (53) [28, 77] 7 (58) [28, 85] PR 4 (20) 4 (24) 3 (25) PD 4 (20) 4 (24) 2 (17)

*<CR at last therapy

или PD при последнем режиме химиотерапии или возобновление заболевания <12 месяцев после аутологичных SCT

or PD on last chemotherapy regimen or disease recurrence <12 months after autologous SCT

[0673] Of the three subjects with DLBCL who are treated with two doses of DL-1 at the time of evaluation, two show a partial response (PR) and 1 shows progressive disease (PD). Among 2 subjects treated with a single dose of DL-2 at the time of evaluation, both subjects were observed to achieve CR. In the MCL cohort with 2 total subjects treated at the time of evaluation with a single dose of DL-1, 1 PR and 1 PD are observed. Two double dose subjects, three triple dose subjects, and four double dose subjects of DLBCL Expressor were observed to achieve a response (7 CR, 2 PR).

[0674] The number of CAR+ T-lymphocytes in peripheral blood is determined at certain time points after treatment using a transgene-specific reagent. Peripheral blood CAR+/CD3+ T-lymphocyte counts measured at specific time points after infusion are shown for subjects grouped by best overall response in FIG. 12A. Responders show higher peak CAR+/CD3+ (CR/PR) T-lymphocyte counts than subjects with progressive disease (PD). Figures 12B-D show the levels of CAR+/CD3+ T-lymphocytes, CAR+/CD4+ T-lymphocytes, and CAR+/CD8+ T-lymphocytes (cells/µl of blood; mean ±SEM) in subjects who achieve response to treatment, grouped by duration of response (continuous response (CR/PR) or PD at 3 months). C _max (CAR+ cells/µl blood) and area under the curve (AUC) for responders (CR/PR) and PD are shown in Table E4. The results are consistent with the finding that persistent responses correlate with higher blood levels of CAR+/CD3 ⁺ T-lymphocytes over time and at peak reproduction.

Table E4. C _max and AUC _0-28 are higher in patients with CR/PR compared with PD CD3 CD4 CD8 CR/PR
(n=16) PD
(n=4) CR/PR
(n=16) PD
(n=4) CR/PR
(n=16) PD
(n=4) C _max (CAR+ cells / µl of blood) Mean value (SD) 612 (1919) 2(1) 220 (754) 1 (0.6) 426 (1314) 0.5 (0.5) Median value (Min, Max) 33 (1, 7726) 1 (1, 3) 8 (1, 3040) 1 (0, 2) 4 (0.5238) 0.3 (0.1) Q1, Q3 7, 123 0.7, 2 2, 46 0.6, 2 0.8, 104 0.1, 0.9 AUC _0-28 Mean value (SD) 5883 (18821) 16 (13) 2369 (8388) 10 (7) 3873 (11963) 6 (6) Median value (Min, Max) 196 (11, 75773) 14 (4, 31) 47 (7, 33740) 9 (3, 17) 23
(1, 47834) 4 (1, 14) Q1, Q3 52, 781 5, 26 16, 261 4, 16 4,761 1, 10

[0675] For one subject with chemo-refractory transformed DLBCL (germinal center subtype with BCL2 rearrangement and multiple copies of MYC and BCL6) administered CAR+ T lymphocytes at DL-1, CD3+/CAR+, CD4+/CAR+ T cell counts, CD8+/CAR+ in peripheral blood measured at specific time points are shown in FIG. 13A. Subject previously treated with them and was refractory to the previous five lines of therapy, including individually titrated etoposide, doxorubicin and cyclophosphamide with vincristine and prednisone plus rituximab (DA-EPOCH-R) and intermediate-intensity allogeneic stem cell transplantation from 8/8 HLA matched unrelated donor. Following allogeneic stem cell transplantation and prior to receiving CAR+ T-lymphocytes, the subject showed 100% donor chimerism in all blood cell lines, ceased immunosuppressive therapy, and had no graft-versus-host disease (GVHD). Prior to administration of CAR+ T-lymphocytes, the subject had an auricular mass and temporal lobe damage observed by positron emission tomography and computed tomography (PET-CT) (Fig. 13B) and confirmed by magnetic resonance imaging ( MRI) (Fig. 13D).

[0676] After receiving treatment With anti-CD19 CAR T-lymphocytes, the subject achieves CR 28 days post-infusion as shown by PET-CT (FIG. 13C) and brain MRI (FIG. 13E), with no observable signs of neurotoxicity or CRS. Three months after the infusion of CAR T-lymphocytes, the appearance of a mass located around the pinna is noted in the subject (Fig. 13F) and an incisional biopsy is performed. As shown in Fig. 13A, after the biopsy, the visible tumor decreased without additional therapy. Pharmacokinetic analysis showed a marked regrowth of CAR+ T-lymphocytes in peripheral blood (to a level above the observed initial proliferation, with peak levels observed approximately 113 days post-infusion), coinciding with tumor regression. The subject then began to achieve a second CR, as confirmed by PET-CT re-staging one month after the biopsy (FIG. 13G), and maintained a CR 6 months after infusion of CAR T-lymphocytes. Additional evaluation of the subject showed that the CNS response is persistent and the subject maintains CR for 12 months.

[0677] These results are consistent with the finding that regrowth and activation of CAR+ T lymphocytes can be initiated in vivo following a decrease in the number or loss of functional or active CAR+ T lymphocytes and/or resume after an antitumor response to therapy with CAR T lymphocytes . Then, after re-propagation in vivo, much later after the initial infusion of CAR+ T-lymphocytes, CAR+ T-lymphocytes are able to resume antitumor activity. This result confirms that replication and activation of CAR+ T lymphocytes can be triggered in vivo and that methods for reactivation of CAR+ T lymphocytes can further enhance their effectiveness.

Example 7 Effects of Lenalidomide on Anti-CD19 CAR T Lymphocyte Activity after Serial Restimulation

[0678] Anti-CD19 CAR+ T lymphocytes generated essentially as described in Example 5 are thawed and incubated with CD19 expressing cells (K562 cells transduced to express CD19) at an effector to target cell ratio (E:T) 2.5:1 in the presence or absence of 1 nM, 5 nM, 60 nM, 550 nM or 5000 nM lenalidomide or in the absence of lenalidomide (control). Target K562-CD19 cells are labeled with NucLight Red (NLR) as described in Example 1 to allow target cells to be tracked by microscopy. Cytolytic activity is assessed by measuring the loss of viable target cells over a period of about 120 hours, as determined by the red fluorescence signal (using the IncuCyte® Live Cell Analysis System, Essen Bioscience). Cells from each condition are inoculated in triplicate. As shown in Figure 14, the results are consistent with the finding that the presence of lenalidomide reduces CAR-mediated cytolytic activity in this assay. In similar assays, results vary with E:T ratio and for different compositions of anti-CD19 CAR+ T lymphocytes (eg generated at different time points and/or from cells from different donors).

[0679] In another study, anti-CD19 CAR+ T lymphocytes were incubated with K562-CD19 effector cells at an E:T ratio of 2.5:1 in the presence of 100 nM or 1600 nM lenalidomide, 2 nM or 166 nM of an alternative compound targeted kinase, vehicle control, or no added compound (CAR-T control). After 120 hours of culture, cells are isolated and evaluated by flow cytometry for surface expression of CD25 or PD-1 in subsets of CD4+ or CD8+ T lymphocytes. As shown in FIG. 15A, incubation of anti-CD19 CAR+ T lymphocytes together with K562-CD19 effector cells in the presence of the highest concentration of lenalidomide (e.g., 1600 nM) results in higher levels of CD25 expression in CD4+ and CD8+ T lymphocytes compared to with other conditions. No differences were observed in surface expression of PD-1 in CD4+ or CD8+ T lymphocytes in the presence of lenalidomide, even at the highest concentration of 1600 nM (FIG. 15B).

[0680] In another study, the amount of IL-10 in culture supernatants was assessed after incubation, for 24 hours, of anti-CD19 CAR+ T lymphocytes together with K562-CD19 effector cells at an effector to target cell ratio (E:T) of 3: 1 or 9:1, in the presence or absence of various concentrations of lenalidomide. As shown in FIG. 16, lenalidomide dose-dependently increases IL-10 secretion and/or accumulation in T-lymphocyte culture supernatants.

Example 8 Effects of Lenalidomide on Anti-CD19 CAR T Lymphocyte Proliferation after Serial Restimulation

[0681] The ability of anti-CD19 CAR+ T lymphocytes to proliferate ex vivo after repeated stimulations is assessed using methods essentially as described in Example 2. Anti-CD19 CAR+ T lymphocytes generated from two donors (pt1 and pt2) essentially the same as described in Example 5, cultured together with irradiated K562 cells transduced to express CD19 (K562-CD19 cells) at a ratio of effector to target cells of 2.5:1 in the presence or absence of 1 μM lenalidomide or 50 nM or 500 nM alternative compound targeting the kinase. For each donor, cells are harvested every 3-5 days for each experimental condition in the wells and counted and restimulated with new target cells using the same culture conditions after re-adjusting the number of cells to the initial inoculation density for each run. Carry out, in General, 4 call stimulation during the 12-day period of cultivation. For each stimulation run, the total number of cells is determined and the results are plotted as the fold change in the number of cells after stimulation (FIG. 17A) or the number of doublings from the initial number (FIG. 17B). As shown in FIGS. 17A and 17B, no change or little effect on anti-CD19 CAR+ T cell proliferation was observed in this restimulation assay when cells were cultured in the presence of lenalidomide compared to no lenalidomide.

[0682] After each return to the beginning after pretreatment, evaluate the cytolytic activity by incubating the re-stimulated cells together with K562-CD19 cells (labeled with NucLight Red (NLR)) at some ratio of effector and target cells (E:T) in the presence of 1 μm lenalidomide or 50 nM or 500 nM alternative compound. Cytolytic activity is assessed by measuring the loss of viable target cells over a period of up to 40-60 hours, as determined by the red fluorescence signal (using the IncuCyte® Live Cell Analysis System, Essen Bioscience). Cells from each condition are inoculated in triplicate. Representative cell death observed at 2nd and 4th restimulation for both donors is shown in Figures 18A (as normalized to K562-CD19-Nuc-labeled cells at t=0) or in Figure 18B (% cell death by compared to the reference carrier medium alone (corresponds to 100%)). As shown in FIGS. 18A and 18B, incubation with lenalidomide was observed to result in a decrease in the cytolytic activity of anti-CD19 CAR+ T-lymphocytes in this assay, under the stimulation conditions tested.

Example 9 Generation of BCMA-Conjugated Beads

[0683] B-lymphocyte maturation antigen (BCMA) is conjugated to the beads by covalently linking a BCMA-Fc fusion polypeptide conjugate containing soluble human BCMA fused at its C-terminus to an IgG Fc region, to the surface of commercially available tosyl-activated magnetic beads (ThermoFisher , Waltham MA). The beads are superparamagnetic, non-porous, monodisperse, tosyl-activated beads that covalently bond primary amino and sulfhydryl groups. Conjugation is performed using beads having a diameter of approximately 2.8 µm (denoted M-280) or 4.5 µm (denoted M-450).

[0684] BCMA-Fc (SEQ ID NO: 22) containing the human BCMA extracellular domain (GenBank No. NP_001183.2) and human IgG1 Fc linked to a linker as follows:

MLQMAGQCSQNEYFDSLLHACIPCQLRCSSNTPPLTCQRYCNASVTNSVKGTNA (BCMA extracellular domain; SEQ ID NO: 18)

GGGGS (linker; SEQ ID NO: 19)

PKSSDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDQFFLYSKLTVDKW

(Human IgG1 Fc; SEQ ID NO: 20.

[0685] A clone encoding a human Fc-BCMA fusion construct with an N-terminal leader sequence of CD33 (SEQ ID NO: 21) is inserted into an expression vector and expressed in HEK 293 cells. The resulting BCMA-Fc fusion protein is determined to be has a purity greater than 95% as assessed by gel permeation chromatography. For binding studies, the BCMA-Fc fusion protein is incubated with T lymphocytes expressing anti-BCMA CARs and T lymphocytes expressing CARs that do not bind to BCMA. Flow cytometry results show that the BCMA-Fc fusion protein specifically binds to CAR-expressing anti-BCMA T lymphocytes.

[0686] Various concentrations of BCMA-Fc fusion protein ranging from 5 μg to 200 μg are added to approximately 1 ml of tosyl-activated beads (for example, containing about 4×10 ⁹ tosyl-activated beads having a diameter of 2.8 μm, or about 4×10 ⁸ tosyl-activated beads having a diameter of 4.5 μm). Covalent binding is carried out by overnight incubation at 37° C. in phosphate buffered saline (PBS) containing 0.1% human serum albumin (HSA). The beads are washed and resuspended in 1 ml PBS with 0.1% HSA. After conjugation, the bead concentration is determined using a Cellometer. In the examples below, BCMA-conjugated beads used in various studies are referred to either by the amount of BCMA-Fc antigen added per ml or by the concentration of antigen (µg/ml) during conjugation, for example, 5 µg or 5 µg/ml ; 50 mcg or 50 mcg/ml; 200mcg or 200mcg/ml and so on.

Example 10 Evaluation of T-lymphocyte markers on anti-BCMA CAR+ T-lymphocytes stimulated with BCMA-conjugated beads in the presence or absence of lenalidomide

[0687] BCMA-conjugated beads (4.5 µm diameter) conjugated to various amounts of BCMA antigen as described in Example 9 are incubated with anti-BCMA CAR+ T lymphocytes in the presence or absence of lenalidomide and expression of T markers is assessed. -lymphocytes.

[0688] Approximately 1.5 x 10 ⁶ CAR+ T lymphocytes are added to the wells of a 12-well plate and incubated with beads from a 200 μg/ml composition of BCMA-conjugated beads at a ratio of CAR+ T-lymphocytes and BCMA-conjugated beads of 1:0, 3, 1:1 or 1:3 (approximately 0.5×10 ⁶ , 1.5×10 and 4.5×10 ⁶ balls per well, respectively). As a control, wells are coated with 5 μg/ml of anti-CD3 antibody (suboptimal concentration for stimulation) or cells are seeded in the absence of any agent (control without stimulation). Each state is incubated in the presence or absence of 5 μM lenalidomide. Cells are incubated for four days and then analyzed by flow cytometry for surface expression of CD4, CD8, Tim3, PD-1, CD25 and CD69.

[0689] As shown in Fig. 19A, the presence of 5 μM lenalidomide increases the proliferative capacity of T lymphocytes (observed by a decrease in the intensity of the CTV dye) after incubation for three days with beads conjugated with 200 μg of BCMA antigen, in the ratio of T lymphocytes and beads 1:1, compared with incubation with beads in the absence of lenalidomide (control carrier medium). As shown in Figures 19B and 19C, the presence of lenalidomide during incubation further increases the extent of CD25 surface expression in CD4+ and CD8+ T lymphocytes induced after incubation of CAR+anti-BCMA T lymphocytes together with BCMA-conjugated beads (FIG. 19B) or stimulation of anti-CD3 (Fig.19C).

[0690] In another experiment, anti-BCMA CAR T-lymphocyte compositions prepared essentially as described in Example 1 were inoculated into 96-well plates at a density of 5 x 10 ⁵ cells per well. The tested CAR T-lymphocyte compositions contain, on average, approximately 45% anti-BCMA CAR+ cells when inoculated. The cells from each composition are incubated for 18 hours in the presence of beads from the composition with 5 μg/ml, 50 μg/ml or 200 μg/ml BCMA-conjugated beads at a ratio of T-lymphocytes to beads 1:1. As a control, cells are incubated with beads conjugated with anti-CD3/anti-CD28 antibody (positive control) or without added agent (negative control). Incubation is carried out without lenalidomide or in the presence of 0.5 μM or 5 μM lenalidomide. After incubation, cells are treated with reagents that allow extracellular and intracellular antibody staining by flow cytometry for the transcription factors Blimp1, EOMES, GATA-3, ikaros, Helios, and Tbet, and the markers CD25, CD31, and PD-1.

[0691] Marker levels after incubation of a CAR+ T cell composition from one exemplary donor are shown for BLIMP-1 (FIG. 20A), CD25 (FIG. 20B), CD31 (FIG. 20C), PD-1 (FIG. 20D), Tbet (Fig.20E) and EOMES (Fig.20F), GATA-3 (Fig.20G) Helios (Fig.20H) and Ikaros (Fig.20I). As shown, the expression of a number of assessed transcription factors and activation markers associated with effector T-lymphocytes increases after stimulation with BCMA-conjugated beads. For many of the markers evaluated, the degree of upregulation is similar to that induced by stimulation with anti-CD3/anti-CD28 beads. In some cases, the degree of stimulation with BCMA-conjugated beads is greatest in the presence of 5 µg beads. As shown in Fig.20I, the level of expression of Ikaros is reduced in the presence of lenalidomide under all conditions. Similar results are observed for the composition of CAR+ T-lymphocytes generated by the second donor, except that there is no change in Helios expression from cells from this donor when stimulated under the conditions studied.

Example 10 Evaluation of Anti-BCMA CAR T Lymphocyte Activity Stimulated with BCMA-Conjugated Beads in the Presence or Absence of Lenalidomide

a. Effector reactions

[0692] Cryo-frozen anti-BCMA CAR+ T lymphocytes prepared essentially as described in Example 2 and prepared at a 1:1 ratio of CD4+ and CD8+ T lymphocytes are thawed. Unless otherwise noted, beads (approximately 4.5 µm diameter from a formulation of 5 µg/mL or 50 µg/mL BCMA-conjugated beads generated as described in Example 9 are added to wells at a ratio of T lymphocytes to beads of 1:1 in the presence or absence of 5 μM lenalidomide Cells are incubated for up to 14 days and analyzed at various time points for cytokine secretion, cell expansion, by flow cytometry against the EGFRt surrogate marker, and for cytolytic activity.

a. Expression of cytokines

(i) Presence of cytokines in the supernatant

[0693] Twenty-four hours after adding BCMA-conjugated beads, assess the presence of TNF-α, IFNγ and IL-2 in the culture supernatants. As shown in Figures 21A-21C, incubation with BCMA-conjugated beads induces the secretion of IFNγ (FIG. 21A), IL-2 (FIG. 21B), and TNF-α (FIG. 21C) in culture supernatants. The rate of cytokine production is higher when cells are incubated with beads from a 50 μg/ml BCMA conjugated bead formulation compared to a 5 μg/ml BCMA conjugated bead formulation, demonstrating that CAR stimulation by BCMA beads is dose-dependent. Lenalidomide has been shown to increase BCMA-induced cytokine production by CAR+ T lymphocytes after stimulation with BCMA-conjugated beads.

[0694] In another exemplary study, two different compositions of anti-BCMA CAR T lymphocytes are generated from different donors, each containing T lymphocytes expressing the same anti-BCMA CAR. Cells are thawed and incubated with beads (approximately 4.5 µm in diameter) from a formulation of 5 µg/ml or 200 µg/ml BCMA-conjugated beads generated as described in Example 1. Incubation is carried out at a ratio of T lymphocytes to beads of 1: 1 in the presence or absence of 1 μM or 5 μM lenalidomide. Twenty-four hours after the addition of the BCMA-conjugated beads, the production of IL-2 by anti-BCMA CAR+ T lymphocytes in the culture supernatants was assessed. As shown in FIG. 21D, higher IL-2 production is observed in the presence of high antigen challenge (200 μg/ml BCMA-conjugated beads) compared to lower antigen challenge (5 μg/ml BCMA-conjugated beads). Lenalidomide, either at 1 μM or 5 μM, increases cytokine production in the presence of both high and low antigen stimulation.

(ii) Intracellular levels of cytokines

[0695] Anti-BCMA CAR+ T-lymphocytes are incubated in the presence of 1 μM lenalidomide or vehicle and 50 μg/ml BCMA-Fc conjugated beads for 2 hours, and cells are evaluated by flow cytometry for phosphorylated STAT 5. To evaluate cytokine levels IFNγ and TNFα, anti-BCMA CAR+ T lymphocytes are incubated in the presence of 0.1 μM or 1 μM lenalidomide or vehicle and 5 μg/ml, 50 μg/ml, or 200 μg/ml BCMA-Fc conjugated beads for 24 hours. The cells are gated on transduced live CD3+ cells and evaluated by flow cytometry for intracellular accumulation of IFNγ and TNFα cytokines in CD4+ and CD8+ cells.

[0696] As shown in FIG. 22A, 2-hour antigen challenge increases the percentage of cells positive for phosphorylated-STAT5 compared to no challenge control (shown by the dotted line). Results for intracellular levels of the cytokines IFNγ and TNFα from anti-BCMA CAR+ T lymphocytes generated from a representative normal CAR T lymphocyte donor are shown in FIG. 22B. In this study, cytokine production by anti-BCMA CAR T lymphocytes is increased by lenalidomide over a wide range of antigen levels and concentrations.

b. cell proliferation

[0697] Total cell counts monitored on day 4 (FIG. 21E) and day 7 (FIG. 21F) increase after stimulation of T lymphocytes with CAR+ anti-BCMA beads from a formulation of 50 μg/ml BCMA-conjugated beads, but not from formulations of 5 μg/ml BCMA-conjugated beads compared to cells present at the start of incubation (dashed line). A small increase in proliferation is observed in cells incubated with 50 µg beads in the presence of lenalidomide on day 7.

[0698] To further evaluate proliferation, cells containing CAR-expressing anti-BCMA T lymphocytes are labeled with CELLTRACE VIOLET (CTV; ThermoFisher Scientific, Waltham MA) as a proliferation marker according to the manufacturer's protocol prior to incubation with BCMA-conjugated beads. Proliferation is assessed by dye dilution using flow cytometry on cells stimulated with beads from a 50 μg/ml BCMA-conjugated bead formulation. Compared to proliferation without lenalidomide, there is a slight delay in proliferation as assessed by diluting CTV in the presence of lenalidomide on day 4 but not day 7 (FIG. 21G).

c. reproduction

[0699] Four days and seven days after the addition of BCMA-conjugated beads, incubated cells are stained with CD4 or CD8 and anti-EGFR antibody to determine the percentage of cells positive for EGFRt as CAR+ surrogate T lymphocytes. At the time of inoculation, 26% of CD4+ cells express anti-BCMA CAR and 39% of CD8+ cells express anti-BCMA CAR as determined by BCMA-Fc staining. The percentage of CD4+ EGFRt+ T lymphocytes increases from about 26% at the start of incubation to over 40% on day 4 (FIG. 21H) and to over 60% on day 7 (FIG. 21I) when cells are incubated in the presence of beads from the 50 μg formulation /ml BCMA-conjugated beads. As shown in FIG. 21I, the percentage of CD8+ EGFRt+ T lymphocytes increased on day 7 from about 38% at the start of incubation to over 60% when cells were incubated in the presence of beads from a 50 μg/mL formulation of BCMA-conjugated beads. Cell proliferation is greatest when cells are incubated in the presence of 50 μg/ml BCMA conjugated beads as compared to 5 μg/ml BCMA conjugated beads. The presence of lenalidomide did not significantly affect the rate of proliferation of CAR+ T-lymphocytes in this study.

d. cytolytic activity

[0700] The cytolytic activity of CAR+ T lymphocytes after incubation with BCMA-conjugated beads was assessed by incubation with target BCMA-expressing cell line RPMI-8226, which is a multiple myeloma cell line BCMA+. After seven days of incubation of CAR+ T lymphocytes with anti-BCMA together with BCMA-conjugated beads (5 µg/mL or 50 µg/mL) in the presence or absence of lenalidomide, the beads are removed from the cultures and the cells are inoculated with target RPMI-8226 cells at the ratio of effector cells to target cells 3:1 or 1:1 in the presence or absence of 5 μm lenalidomide in the future. To perform cytolytic analysis, RPMI-8226 target cells are labeled with NucLight Red (NLR) to allow target cells to be tracked by microscopy. Cytolytic activity is assessed by measuring the loss of viable target cells over a period of four days, as determined by the red fluorescence signal (using the INCUCYTE® Live Cell Analysis System, Essen Bioscience). Viable cell counts are normalized per cell on day 0 prior to incubation with target RPMI-8226 cells.

[0701] Exemplary results for a 1:1 ratio of effector to target cells are shown in FIG. 21J. As shown, anti-BCMA CAR+ T lymphocytes show efficient killing in this assay. CAR+ anti-BCMA T cells stimulated with 5 μg/mL BCMA conjugated beads are slightly less effective at killing cells than CAR+ anti-BCMA T cells stimulated with 50 μg/ml BCMA conjugated beads . For all conditions, pre-incubation with lenalidomide for a seven-day incubation prior to the kill assay increased the cytolytic activity of CAR+ T-lymphocytes. The presence of lenalidomide during the cell killing assay does not significantly affect the killing activity. Cell killing is not observed when RPMI 8226 cells are cultured alone or in the presence of lenalidomide, demonstrating that lenalidomide does not directly affect the viability of target cells in this assay.

B. Serial restimulation

[0702] Anti-BCMA CAR T-lymphocyte compositions were generated from three different donors, each containing T-lymphocytes expressing the same anti-BCMA CAR, thawed and incubated for seven days along with beads (approximately 4.5 µm) at a 1:1 bead to cell ratio from a 50 µg/ml BCMA-conjugated bead formulation generated as described in Example 9. Incubation is carried out in the presence of 5 µM lenalidomide or in the absence of lenalidomide (carrier control medium). Cells are harvested after 7 days and re-inoculated for three additional runs up to 28 days, each run includes a return to the beginning, to the initial seeding density and incubated for an additional 7 days in the presence of the same concentration of lenalidomide.

[0703] At each return to the start after pretreatment, cytolytic activity is assessed by incubating with BCMA-expressing target cell line RPMI-8226 (labeled with NucLight Red (NLR)) at an effector to target cell ratio (E:T) of 1:1 in the presence or absence of lenalidomide in the future. Cytolytic activity is assessed by measuring the loss of viable target cells over a period of up to 80-150 hours, as determined by the red fluorescence signal (using the IncuCyte® Live Cell Analysis System, Essen Bioscience). Cells from each condition are inoculated in triplicate. Determine the % killing of cells compared to the control carrier medium itself (corresponding to 100%).

[0704] FIG. 23A shows the results for cytolytic activity of anti-BCMA CAR+ T lymphocytes from an exemplary donor after pretreatment for 7 days, 14 days, or 21 days. As shown, anti-BCMA CAR+ T lymphocytes that are pre-incubated with lenalidomide for 7 days or 14 days show higher cytolytic activity compared to cells that are not pre-incubated with lenalidomide. In this donor, an overall decrease in killing efficiency is observed for CAR+ anti-BCMA T lymphocytes that are pre-incubated with lenalidomide for 14 or 21 days compared to day 7. The donor exhibits similar effects on cytolytic activity of CAR+ anti-BCMA T lymphocytes. -BCMA after pre-treatment with lenalidomide for 7 or 14 days; cytolytic activity after 21 days after pre-treatment with lenalidomide in this donor is not evaluated. As shown in FIG. 23B, increased killing efficiency in CAR+ T lymphocytes by anti-BCMA is observed for this donor, for those cells pre-incubated with lenalidomide at all time points.

Example 12 Effects of Lenalidomide on PD-1 Expression and PD-L1 Signaling

[0705] Anti-BCMA CAR T lymphocytes were generated from samples of material obtained from representative healthy donors or from a patient with multiple myeloma and cultured with 50 μg/ml BCMA-Fc conjugated beads (generated as described in Example 9) at ratio of balls : T-lymphocytes CAR+ 1:1 for 7 days, in the presence of 1 μm lenalidomide or control carrier medium. Expression of CD25, PD-1, Tim3 and Lag3 on CAR T-lymphocytes (using an antibody instead of a CAR surrogate marker) cultured under various conditions is then assessed by flow cytometry.

[0706] Such anti-BCMA CAR T lymphocytes pre-stimulated with beads in the presence or absence of lenalidomide or freshly thawed anti-BCMA CAR T lymphocytes generated from comparable donor samples are then separated from the beads, washed and cultured together with the target RPMI-8226 cells (labeled with NucLight Red (NLR) to allow them to be tracked by microscopy) in the presence of 1 μM lenalidomide or vehicle control. Specifically, for pretreated cells pretreated in the presence of lenalidomide, the cells are cultured together with the target cells in the presence of lenalidomide; similarly, for pretreated cells that are pretreated in the presence of a carrier medium, the cells are cultured together with target cells in the presence of a carrier medium. After co-culture, cytolytic activity is assessed by measuring the loss of viable target cells over a period of seven days, as determined by the red fluorescence signal. The kill rate is normalized to CAR+ anti-BCMA T-lymphocytes previously stimulated on the beads in the presence of vehicle. Cytokine production is assessed by ELISA from the supernatant after culturing with target cells for 24 hours. The experiments are carried out twice on 3 donors. Linear fixed exposure models or mixed exposure models are used to assess the significance of lenalidomide treatment for cytolytic activity and cytokine production, treating treatment, donor, and time as fixed exposures and the animal as random exposure nested by time when repeated measurements obtained from the same animal. The P values are obtained by probability ratio tests comparing the full model with the impact of interest and the model without the impact of interest.

[0707] Figure 24A shows results for antigen-specific CAR cytolytic activity and Figure 24B shows results for cytokine production for anti-BCMA CAR T lymphocytes that are prestimulated with BCMA beads, (versus freshly thawed (not prestimulated) T-lymphocytes CAR anti-BCMA) in cocultures comparing cells cultured in the presence and absence of lenalidomide. Prestimulated CAR T lymphocytes show reduced cytolytic activity (P=2.1×10 ^-4 ) and cytokine production (P=0.03 for IFN-γ) compared to freshly thawed anti-BCMA CAR+ T lymphocytes. In the absence of lenalidomide in pretreatment and post-coculture, prestimulated CAR T lymphocytes show reduced cell killing and cytokine production compared to fresh CAR T lymphocytes, indicating that chronic prestimulation leads to functional impairment. These results are consistent with a wasted-like phenotype induced by pre-stimulation on BCMA-conjugated beads. The presence of lenalidomide during the pre-stimulation period retains cytolytic function (P=0.04) and there is a trend towards increased cytokine production compared to cells exposed to vehicle during the pre-stimulation period (FIG. 24B). The presence of lenalidomide in this assay is consistent with the observation that lenalidomide can reduce the effects indicative of a wasting-like phenotype in pre-stimulated CAR T lymphocytes.

[0708] As shown in Figure 24C, the phenotype of anti-BCMA CAR+ T lymphocytes stimulated for 7 days on BCMA beads is assessed, and the addition of lenalidomide significantly increases the viability of CAR+ anti-BCMA CAR T lymphocyte material for 3 healthy donors (P =0.04). The addition of lenalidomide did not change the total cell count for all donors during this 7 day period, and there was no significant difference in percentage of CAR+ between CAR T cells treated with vehicle and lenalidomide. 24D shows representative flow cytometric analysis of CD25 and PD-1 surface expression (mean fluorescence intensity (MFI), for anti-BCMA CD4+ or CD8+ CAR T-lymphocytes after stimulation (pre-treatment) with BCMA beads for 7 days, in the presence of or in the absence of 1 μM lenalidomide As shown, the results indicate that lenalidomide reduces PD-1 expression by CAR BCMA T lymphocytes while increasing CD25 expression after prolonged stimulation As shown in Figure 24E, flow cytometric analysis for three donors CAR T shows that the addition of lenalidomide increases the surface expression of Tim3 in the CD8+ population (P=4.0 × 10-4), with mixed effects on the CAR+ CD4+ population.Among all donors and in both CAR+ populations, both CD4+ and CD8+, lenalidomide increases the level of CD25 (CD4+ and CD8+; P=2.2 × 10 ^-16 ) and the percentage positive for Lag3 expression (CD8+, P<0.03; CD4+, P=0.002). Note that there is also a decrease in the percentage of PD-1+ cells in the CD4+ population (P=0.04), with 2 out of 3 donors also showing a decrease in the CD8+ population.

[0709] In another study, recombinant human BCMA-conjugated beads were used to stimulate CAR T lymphocytes at various concentrations to titrate the amount of stimulation, either low (5 µg/mL), medium (50 µg/mL), or high (200 mcg/ml) stimulation. Under conditions of medium stimulation, the production of secreted cytokines is measured 24 hours after stimulation and a 200% increase in IL-2 and TNF-α concentrations compared to vehicle control is observed, with a donor-dependent increase in IFN-γ levels (FIG. 25A). Cells are stimulated with BCMA-conjugated beads for 24 hours in the presence of 0.1 μM or 1.0 μM lenalidomide or vehicle control. A protein transfer inhibitor is added during the last hours of incubation and cells are stained for intracellular IL-2, IFN-γ and TNF-α.

[0710] Anti-BCMA CAR+ T lymphocytes activated on BCMA beads show a stimulation level-dependent effect on cytokine production, with 5 μg BCMA beads eliciting limited production of CAR T effector cytokines compared to 50 μg and 200 μg BCMA beads (Fig. 25B). Lenalidomide increases the percentage of intracellular IFN-γ ⁺ and TNF-α ⁺ staining at all levels of stimulation for CAR T-lymphocytes, both CD4+ and CD8+. The amount of stimulation either raises or lowers IL-2 in response to lenalidomide, with lenalidomide decreasing the percentage of CAR+ IL-2 ⁺ T cells at 50-ug and 200-ug at stimulation, but increasing the percentage of CAR+ IL-2 ⁺ T cells under conditions of 5-mcg stimulation. In the absence of stimulation, lenalidomide has no effect on CAR T cytokine production, indicating that the enhancement of cytokine production provided by lenalidomide requires stimulation.

[0711] In another study, to see if lenalidomide-induced potentiation of CAR T activation and cytokine production overrides PD-L1-mediated inhibition, cells were cultured in the presence of BCMA beads generated as described in Example 9, with additional conjugation of recombinant PD- L1-Fc human or without. CAR T-lymphocytes obtained from healthy donors or patients are stimulated in the presence of BCMA-conjugated beads or BCMA/PD-L1-conjugated beads for 24 hours in the presence of 1 μM lenalidomide. The production of cytokines in the supernatant is measured. The results are shown in Fig.25C. As shown in FIG. 25C, scores of CAR T lymphocytes from both healthy donors and patients demonstrate that the addition of recombinant PD-L1 to recombinant BCMA beads reduces IFN-γ, IL-2, and TNF-α levels. Treatment with lenalidomide has been shown to potentiate levels of secreted cytokines above those from CAR T-lymphocytes treated with vehicle in the presence of PD-L1. The results are consistent with the conclusion that the production of anti-BCMA CAR-T cytokines after incubation with BCMA-conjugated beads increases lenalidomide levels in the presence of PD-L1-mediated inhibition.

Example 13 Analysis of gene expression and chromatin availability in CAR T-lymphocytes in the presence or absence of lenalidomide

[0712] Gene expression and chromatin availability in CAR T-lymphocytes are assessed when stimulated in the presence or absence of lenalidomide. Anti-BCMA CAR-expressing T-lymphocytes generated from four (4) different independent donors stimulated with 50 μg/ml BCMA-conjugated beads for 24 hours (24 hours+stim) or 7 days (d7+stim), or cultured without stimulation for 24 hours (24 hours), with or without lenalidomide (1 μM). Experiments are carried out twice on 3-4 donors. CAR-expressing cells are assessed by RNA sequencing (RNA-seq) for gene expression and analyzed for transposase-accessible chromatin using chromatin accessibility sequencing (ATAC-seq). Analyzes are performed on 50,000 cells at each time point.

[0713] RNA-seq is performed on complementary DNA samples (cDNA) prepared from RNA isolated from cultured CAR-expressing anti-BCMA cells. ATAC-seq is carried out, in general, as described in Buenrostro et al., Nat Methods. (2013) 10(12): 1213-1218. Paired-end ATAC read fragments are cut, aligned with Bowtie2, and filtered for quality, fragment length, duplication, and mitochondrial contribution. ATAC-seq availability peaks are determined using MACS2 (q<0.01) and a consensus panel is generated from overlapping peaks present in 2 or more samples using DiffBind. Principal Component Analysis (PCA) is performed on RNA-seq and ATAC-seq datasets generated from DESeq2 normalized samples. Calculate differential expression (DE, for RNA-seq) or consensus peak availability (DA, for ATAC-seq) simulating the effects of donors (Donors 1-4) and treatment effects (lenalidomide versus vehicle) at 24 hours and per day 7. The differential locus selection threshold corresponds to q≤0.05 and log2-fold change ≥0.5 for RNA-seq or q≤0.1 for ATAC-seq. Gene Ontology (GO) enrichment analysis is performed and activation z-score is determined on a subset of genes differentially expressed at q<0.1 using Ingenuity Pathway Analysis software (Qiagen, Inc.), considering donor exposures within each treatment condition . Motif enrichment analysis is performed for peaks shown to be more accessible in the presence of lenalidomide using the HOMER software, using the peak consensus set as background, for the ATAC-seq data at day 7 stimulation (d7+stim).

[0714] RNA-seq heatmaps are generated from normalized (transcripts per million) expression data averaged across donors per condition and normalized by row using z-scores. Motif enrichment in DA peaks on day 7 was performed with HOMER using consensus peak sets as background.

[0715] PCA results representing the overall diversity of gene expression or chromatin availability on the genome are shown in Fig. 26A (gene expression; based on RNA-seq results) and Fig. 26B (chromatin accessibility; based on ATAC-seq results). Ellipses are drawn to indicate such groups, as it is observed that the main factors that contribute to the scatter of gene expression or chromatin availability are culture time and the presence of stimulation. Cells cultured in the presence of lenalidomide (circles) show different overall gene expression and chromatin availability compared to cells cultured in the absence of lenalidomide (triangles, vehicle), they show the impact of lenalidomide treatment for each donor and culture conditions. For lenalidomide treatment, the overall direction of change (shown by the dotted line between the triangle and circle) is similar for each donor, and the degree of change is generally greater in cells cultured for 7 days on challenge compared to those in cells cultured in within 24 hours, with or without stimulation. Thus, PCA shows clustering based on stimulation (stim or no stim) and time (24 hours or 7 days) for both RNA-seq (Figure 26A) and ATAC-seq (Figure 26B) datasets.

[0716] The role of lenalidomide is then examined at 24 hours or 7 days of stimulation after accounting for variation between donors. Figures 27A-27D show changes in gene expression (Figures 27A and 27B, after 24 hours and 7 days of stimulated culture, respectively) or chromatin availability (Figures 27C and 27D, after 24 hours and 7 days of stimulated culture, respectively) in the presence of lenalidomide. RNA-seq analysis shows up-regulation of a small set of genes (214) after 24 hours, and in the presence of lenalidomide more genes (583) are changed after 7 days of stimulation (Figures 27A and 27B). ATAC-seq analysis shows a limited set of changes in chromatin availability associated with lenalidomide treatment after 24 hours of stimulation, with dramatic profile changes and an increase in the number of sites with changes in chromatin accessibility (changes in chromatin availability in 2804 peaks) after 7 days of stimulation in the presence of lenalidomide (Figures 27C and 27D). These results indicate that treatment with lenalidomide alters both the transcriptional and epigenetic profile of CAR T lymphocytes.

[0717] To further identify specific transcriptional changes associated with lenalidomide treatment, gene ontology analysis is applied to the RNA-seq dataset and identifies biological signaling pathways that are enriched in differentially expressed genes (Figures 28A and 28B). The direction and significance of the effects on biological pathways are shown after 24 hours (FIG. 28A) or after 7 days (FIG. 28B). The results show that the presence of lenalidomide results in an increase in the expression of genes involved in the activation and signaling of T-lymphocytes. The results show that pathways differentially regulated in the presence and absence of lenalidomide show gene enrichment in immune synapse-associated genes involved in cytokine signaling and genes involved in T-lymphocyte activation pathways. Specifically, pathways associated with T-lymphocyte chemotaxis (leukocyte extravasation, integrin, ILK, and CXCR4-associated gene sets), intracellular signaling, and the cytoskeleton (Rac/Rho/Cdc42) are up-regulated in the presence of lenalidomide within 24 hours of stimulation compared to with control carrier medium. In addition, these data support an increase in ICOS-related signaling pathways - data that is consistent with previous publications demonstrating an increase in ICOS and ICOSL levels in a population of CD3+ peripheral blood mononuclear cells treated with ex vivo lenalidomide (Gorgun et al. (2010) Blood, 116:3227-3237). After 7 days of stimulation, lenalidomide up-regulates pathways associated with Th1 T-lymphocyte response and co-stimulation, while at the same time reducing the signatures of Th2-associated genes.

[0718] For a selected subset of genes, including genes involved in T-lymphocyte activation and signaling, changes in gene expression and chromatin availability in the presence of lenalidomide are compared for cells cultured for 7 days on stimulation to determine if chromatin availability correlates with transcription. 29 shows the individual chromatin availability peaks (diamonds) and the average change in chromatin availability for each gene (circles) in a plot against the respective changes in gene expression measured by RNA-seq showing signal agreement in the two methods. For donors, a significant increase in chromatin availability is observed among multiple loci associated with IFN-γ and IL-2RA (CD25), and these changes correlate with a significant increase in transcription. Importantly, the up-regulation of IFN-γ and CD25 supports previous data from chronic stimulation experiments. In addition, a decrease in chromatin availability and gene transcription in CD69 and CCR7 is also observed upon treatment with lenalidomide.

[0719] The analyzed ATAC-seq dataset for motif enrichment and the results of motif enrichment analysis for peaks with increased availability in the presence of lenalidomide in 7 day old cultures are shown in FIG. Motifs predicted to bind various transcription factors understood to be involved in T-lymphocyte activation and signaling, including AP-1/Jun and nuclear factor κB, are enriched in peaks with increased availability in the presence of lenalidomide. The results are consistent with an increase in the functional activity of CAR-expressing T-lymphocytes in the presence of lenalidomide.

[0720] Without wishing to be bound by theory, RNA- and ATAC-seq studies result in a number of insights regarding possible mechanisms for lenalidomide-induced increases in CAR T function. First, the number of transcriptional changes and changes in chromatin availability associated with stimulation and time are compared mainly with the effects of lenalidomide, indicating relatively subtle effects of lenalidomide on transcriptional networks. Second, the changes associated with lenalidomide are very broad, including early transcriptional changes associated with cytoskeletal repair and chemotaxis. After chronic stimulation, a distinct transcriptional signature appears that includes a decrease in transcripts associated with the Th2 response, the G2/M checkpoint, and ATM, along with an increase in Th1, peroxisome proliferator-activated receptor γ, and genes associated with the actin cytoskeleton. These effects may support the role of lenalidomide treatment and cell cycle control and T-lymphocyte activation. Previous studies have also demonstrated the effects of IMiD on Th1- and Th2-associated signatures, as well as changes in elements associated with cytoskeletal repair and T-lymphocyte migration. Demonstrated early changes in cytokine production under the influence of lenalidomide can contribute to the change in the state of T-lymphocytes, that is, they can enhance both memory function and effector function at the same time. Overall, these results suggest that additional factors other than those previously reported are involved in lenalidomide-induced prolongation of CAR T function, including possible alterations in cell cycle control.

[0721] The use of ATAC-seq provides additional insight into the potential mechanisms of action of lenalidomide. Although both stimulation and timing are one of the main drivers of changes in chromatin availability, treatment with lenalidomide is associated with an increase in chromatin availability at loci enriched in motifs associated with T-lymphocyte activation and function after chronic stimulation. These epigenetic changes are consistent with the observed functional changes in CAR T-lymphocytes incubated with lenalidomide. Changes in chromatin accessibility signatures are associated with T-lymphocyte depletion and may represent a more robust indicator of depletion compared to surface expression of T-lymphocyte ligands. These data demonstrate that chronic stimulation with lenalidomide results in an increase in chromatin availability and gene expression of IL-2 and CD25 and a decrease in gene expression and availability of CCR7 and CD69 chromatin. Previous studies suggest that CCR7-expressing cells produce higher levels of IL-2; however, current research indicates that the IL-2 pathway could be independently altered by lenalidomide, resulting in an alternate state of the T-lymphocyte. CD69, a T-lymphocyte activation marker, contains a nuclear factor κB responsive element that is required for the CD69 response to TNF-α. Closure of CD69-associated chromatin and reduction in transcripts may be a response to a long-term increase in TNF-α production by CAR T lymphocytes co-cultured with lenalidomide, or it may be a response of T lymphocytes to increased activation in the presence of lenalidomide. Cells treated with lenalidomide show an increase in transcription factor motif enrichment for factors associated with T-lymphocyte activation, supporting the idea that these cells are exposed to long-term activation signaling. In general, the lenalidomide-induced CAR T-lymphocyte state includes elements such as effector T-lymphocyte functions, including both increased IFN-γ and TNF-α production, and T-lymphocyte function, including increased IL-2 levels and long-term proliferation.

Example 14 Evaluation of the pharmacodynamic response of transcription factor Ikaros in CAR-expressing T-lymphocytes in the presence of compound 1 or lenalidomide

[0722] T-lymphocyte compositions containing CAR-expressing anti-CD19 T-lymphocytes are generated from leukapheresis samples from three healthy adult human donors by a method comprising selecting T-lymphocytes based on immunoaffinity (including CD4+ and CD8+ cells) from the samples resulting in two compositions enriched in CD8+ and CD4+ cells, respectively. Cells are incubated in the presence of 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione (Compound 1) or lenalidomide and expression of the Ikaros transcription factor is evaluated.

[0723] Cells of compositions enriched in CD4+ and CD8+ are activated individually with anti-CD3/anti-CD28 beads and subjected to lentiviral transduction with a vector encoding anti-CD19 CAR. The anti-CD19 CAR contains an anti-CD19 scFv derived from a mouse antibody (variable region derived from FMC63), an immunoglobulin derived spacer, a transmembrane domain derived from CD28, a costimulatory region derived from 4-1BB, and a CD3 intracellular signaling domain. -zeta. The expression construct in the viral vector further contains sequences encoding a truncated receptor that serves as a surrogate marker for CAR expression, which is released from the CAR sequence by the T2A ribosomal skipping sequence. The transduced populations are then individually incubated in the presence of stimulatory reagents for cell expansion. Expanded CD8+ and CD4+ cells are prepared and cryopreserved separately and stored. Cryopreserved CAR-expressing anti-CD19 CD4+ and CD8+ cells from each donor are thawed and pooled at a CAR+ CD4+:CD8+ ratio of approximately 1:1 before use.

[0724] Approximately 2.5 x 10 ⁵ cells (CD4+ and CD8+ T lymphocytes pooled at a ratio of 1:1) from the generated CAR+ T lymphocyte composition are stimulated overnight with a CAR specific reagent and then the cells are incubated with lenalidomide ( 100 nM - 10000 nM), Compound 1 (10 nM - 3000 nM) or vehicle control overnight at 37° C., 5% CO ₂ . The estimated concentrations of Compound 1 and lenalidomide cover the known clinical C _max and C _min . After incubation, CAR-expressing anti-CD19 T lymphocytes are stained with antibodies and analyzed by flow cytometry to evaluate surface expression of CD4, CD8 and a surrogate marker for CAR expression, and intracellular levels of Ikaros in CAR+ CD4+ or CAR+ CD8+ cells. Median fluorescence intensity (MFI) values for Ikaros are normalized and calculated as a percentage relative to the control carrier medium.

[0725] As shown in FIG. 31, a concentration-dependent decrease in expression of intracellular Ikaros is observed in both anti-CD19 CD4+ CAR-expressing T lymphocytes and anti-CD19 CD8+ CAR-expressing T lymphocytes after incubation with Compound 1 or lenalidomide. A greater decrease in Ikaros expression is observed in cells in the presence of Compound 1 compared to lenalidomide. The EC50 value for reducing Ikaros expression is calculated as determined by the concentration of inhibitor that reduces the MFI of Ikaros to 50% of its maximum MFI in the absence of inhibitor. The EC50 values for Compound 1 and lenalidomide are shown in Table E5.

Table E5. EC50 (nM) of Ikaros in CAR+ CD4+ T-lymphocytes and CAR+ CD8+ T-lymphocytes.

CD4+ CD8+ Lenalidomide Compound 1 Lenalidomide Compound 1 Donor 1 61.2 67 80.9 100.9 Donor 2 ND 41.5 ND 60.8 Donor 3 169.8 99.8 235.5 161.1

ND=not defined

Example 15 Evaluation of functional effects on CAR-expressing T-lymphocytes after incubation with target cells in the presence of Compound 1 or lenalidomide

[0726] Anti-CD19 CAR-expressing T-lymphocyte compositions (containing CD4+ and CD8+ cells combined at a 1:1 ratio) were generated essentially as described in Example 14 and incubated with human CD19 (K562.CD19) transduced target K562 cells in the presence of 3-(5-amino -2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione (Compound 1) (at a concentration of 10 nM, 100 nM, 500 nM and 1000 nM), lenalidomide (at a concentration of 100 nM, 1000 nM and 10000 nM) or control vehicle at 37°C, 5% CO₂. Cytokine expression, cytolysis of target cells, and expression of surface markers by CAR-expressing anti-CD19 T lymphocytes are assessed.

A. Cytokine production

[0727] To evaluate cytokine production, 1×10 ⁵ CAR+ anti-CD19 cells (CD4+ and CD8+ T lymphocytes pooled at a ratio of 1:1) are incubated with target K562.CD19 cells at an E:T ratio of 5:1 or 2 ,5:1 in the presence or absence of lenalidomide or Compound 1 as described above. After 24 hours, the supernatants are harvested and analyzed for the production of the cytokines IFN-γ, IL-2 and TNF-α.

[0728] As shown in Figures 32A (Compound 1) and 32B (lenalidomide), cytokine production is increased in the presence of Compound 1 or lenalidomide, respectively, in a concentration dependent manner compared to vehicle control, with an E:T ratio of 5: 1 and 2.5:1. There are differences in cytokine levels among different donors. Treatment with Compound 1 results in greater cytokine production for a variety of conditions compared to treatment with lenalidomide at equivalent concentrations. The increase is statistically significant, as determined by the increase in cytokine production in the presence of 100 nM and 1000 nM Compound 1 compared to the equivalent concentration of lenalidomide at an E:T ratio of 2.5:1 and 5:1, as determined using an unpaired parametric t-test with Welsh correction, see Table E6 (P-values at E:T 2.5:1/P-values at E:T 5:1).

[0729] Table E6. Cytokine production by CAR-expressing anti-CD19 T lymphocytes treated with Compound 1 or lenalidomide.

Donor 1 Donor 2 Donor 3 Concentration (nM): 100 1000 100 1000 100 1000 IFN-γ ***/* */ns ns/ns ns/*** ***/*** ***/*** IL-2 **/* */* **/ ns/ns ns/* ns/ns TNF-α ***/* ***/*** **/ ***/*** **/* **/**

p≤0.05 : *; p ≤0.01: **; p≤0.001: ***; ns: not significant

B. Cytolytic function

[0730] To evaluate cytolytic function, 1×10 ⁵ or 5×10 ⁴ anti-CD19 CAR+ cells (CD4+ and CD8+ T lymphocytes pooled at a ratio of 1:1) are incubated with 2×10 ⁴ target K562.CD19 cells at an E:T ratio of 5:1 or 2.5:1 in the presence or absence of lenalidomide or Compound 1 or vehicle control as described above. Target K562.CD19 cells are transduced with NucLight Red to allow their tracking by microscopy. Cytolytic activity is assessed by measuring the loss of viable target cells over a period of five days, as determined by the red fluorescence signal (using the IncuCyte® Live Cell Analysis System, Essen Bioscience). The kill rate is determined using the formula: 1/AUC and the kill rate is normalized to CAR+ cells co-cultured with target cells that are incubated with vehicle control (established at 100% kill).

[0731] As shown in Figure 33, Compound 1 and lenalidomide generally have a limited effect on the cytolytic function of anti-CD19 CAR-expressing T lymphocytes. When CAR is stimulated in the presence of a higher amount of antigen, as present in cocultures containing an E:T ratio of 2.5:1, Compound 1 and lenalidomide slightly decrease the cytolytic activity of CAR-expressing anti-CD19 cells in the same donors. When CARs are stimulated in the presence of less antigen, as present in cocultures containing an E:T ratio of 5:1, there is a slight but consistent increase in the cytolytic activity of CAR-expressing anti-CD19 T-lymphocytes against target cells compared to cells that are incubated in the presence of high concentrations of Compound 1 or lenalidomide for donor 2, while no effects were observed for donors 1 and 3.

C. Expression of T cell surface markers

[0732] To assess the surface expression of various T lymphocyte markers, 1×10 ⁵ target K562.CD19 cells are incubated with anti-CD19 CAR+ cells (CD4+ and CD8+ T lymphocytes pooled at a ratio of 1:1) at an E:T ratio of 5 :1 or 2.5:1 in the presence or absence of lenalidomide or Compound 1 or vehicle control as described above. After 24 hours, CAR-expressing T lymphocytes stain for CD3, CD4, CD8 and a surrogate marker for CAR expression, as well as the following surface markers: CD69, CD107a, PD-1, CD25, CD62L, CCR7, CD45RO, CD27, and LAG3 .

[0733] Expression levels of selected markers on CAR-expressing CD4+ T-lymphocytes and CAR-expressing CD8+ T-lymphocytes change, as a rule, less than twice, in relation to cocultures with a control carrier medium. Changes in marker expression in the presence of lenalidomide or Compound 1 are donor-dependent, although for the memory markers assessed, CD45RO expression is increased and CD27 is decreased for all donors and E:T ratios. Expression of CD27 is down-regulated in a concentration dependent manner in response to Compound 1 or lenalidomide. Expression of CD69 and LAG3 increased in a concentration dependent manner for cells derived from donor 3 after incubation with Compound 1 but not after incubation of CAR+ cells derived from the same donor with lenalidomide. Expression of the other activation markers assessed remains unchanged in donors treated with lenalidomide or Compound 1. The results are consistent with the observation that Compound 1 and lenalidomide have the potential to intrinsically modulate the early activation phenotypes of CAR-expressing T lymphocytes.

Example 16 Evaluation of cytokine production and surface marker expression by CAR-expressing T-lymphocytes after stimulation with an anti-idiotypic antibody in the presence of Compound 1 or lenalidomide

[0734] Assays similar to those described in Example 15 were performed to assess cytokine production and surface marker expression following CAR-dependent stimulation of CAR-expressing T lymphocytes in the presence of or 3-(5-amino-2-methyl-4-oxo-4H- quinazolin-3-yl)-piperidin-2,6-dione (Compound 1) or lenalidomide, except that CAR-expressing cells are stimulated by an anti-idiotypic antibody. Anti-idiotypic antibody is used to simulate different levels of stimulation in cocultures, which is generally not possible with target K562.CD19 cells because antigen expression is uniformly high on K562.CD19 cells.

[0735] Compositions of anti-CD19 CAR expressing T lymphocytes (comprising CD4+ and CD8+ T lymphocytes combined at a ratio of 1:1) are generated essentially as described in Example 14. Approximately 1×10 ⁵ CAR expressing cells are added in the wells of a 96-well plate, which is pre-coated with an anti-idiotypic antibody specific for scFv CAR-expressing anti-CD19 T-lymphocytes at a concentration of 0, 0.3, 3 and 30 μg/ml. Cells are cultured in the presence of Compound 1 (at 100 nM and 1000 nM), lenalidomide (at 500 nM and 5000 nM) or vehicle control at 37° C., 5% CO ₂ . The expression of cytokines and the expression of surface markers of CAR-expressing anti-CD19 T-lymphocytes are assessed.

A. Cytokine production

[0736] Stimulated culture supernatants are harvested 24 hours later and analyzed for cytokine production. In Figures 34A and 34B, the level of cytokine production in the absence of Compound 1 or lenalidomide (carrier control) is shown with a dashed line. As shown in Figures 34A and 34B, the production of IFN-γ, IL-2 and TNF-α is increased compared to the control vehicle after treatment with Compound 1 or lenalidomide, respectively. The increase is especially evident at an intermediate level of stimulation with 3 kg/ml anti-idiotypic antibody.

B. Expression of surface markers of T-lymphocytes

[0737] Surface marker expression on CAR-expressing anti-CD19 T lymphocytes was assessed after 4 days in culture at various concentrations of anti-idiotypic antibody in the presence of Compound 1 or lenalidomide. CAR-expressing T lymphocytes are stained for CD3, CD4, CD8 and a surrogate marker for CAR expression, as well as for the following markers: CD25, PD-1 and CD69.

[0738] Figures 35A and 35B show the expression of a cell surface marker on CAR-expressing CD4+ and CD8+ cells, respectively, on cells stimulated in the presence of Compound 1, and Figures 36A and 36B show the expression of a cell surface marker on CAR-expressing CD4+ and CD8+ cells , respectively, on cells stimulated in the presence of lenalidomide. In the figures, the expression level of the surface marker in the absence of Compound 1 or lenalidomide (carrier control) is indicated by a dashed line. As shown, an increase in the level of surface markers CD25 and CD69 is observed in CAR-expressing CD4+ and CD8+ T lymphocytes in the presence of Compound 1 or lenalidomide in some donor-derived CAR-expressing cells, and it depends on the amount of stimulation with CAR. Expression of PD-1 is also increased in the presence of Compound 1 or lenalidomide in cells generated from at least one donor, although PD-1 levels are unchanged or decreased in cells generated from other donors. An increase in the expression of a surrogate marker for CAR expression is observed after the addition of Compound 1 or lenalidomide at a suboptimal dose of anti-idiotypic antibody of 0.3 μg/ml, but at higher concentrations of anti-idiotypic antibody, expression of the surrogate marker remains unchanged or decreases.

Example 17 Evaluation of surface marker expression and expansion potential of CAR-expressing T-lymphocytes after serial stimulation in the presence of Compound 1 or lenalidomide

[0739] Serial stimulation of CAR-expressing T-lymphocytes with anti-CD19 is performed to assess the short-term and long-term effects of 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6 -dione (Compound 1) and lenalidomide. Compositions of CAR-expressing anti-CD19 T-lymphocytes (containing CD4+ and CD8+ T-lymphocytes combined at a ratio of 1:1) are generated essentially as described in Example 14. Generated combinations of CAR+ T-lymphocytes are added as 1×10 ⁵ cells per well in a 96-well plate and incubated with irradiated red positive K562.CD19 target cells in the presence of Compound 1, lenalidomide or vehicle control at 37° C., 5% CO ₂ at two different ratios of effector to target cells (E: T) 10:1 and 2.5:1. Incubation is carried out in the presence of Compound 1 (10, 100 or 500 nM), lenalidomide (100 or 1000 nM) or vehicle control. Cells are counted every 3-4 days (starting with each new entry). Cells are then harvested and re-inoculated at 1×10 ⁵ CAR-expressing anti-CD19 cells with fresh media, freshly added Compound 1 or lenalidomide at the same concentration, and with freshly irradiated target K562.CD19 cells. This is repeated for 7 rounds of serial stimulation and cells are evaluated at various time points for surface marker expression, cytolytic activity, and expansion potential.

A. Expression of surface markers

[0740] Expression of selected surface markers is assessed on cells at day 4 (ie, 4 days after the first stimulation) and day 28 (ie, 4 days after the seventh stimulation). Specifically, harvested cells are analyzed by flow cytometry for CD3, CD4, CD8 and a surrogate marker for CAR expression, as well as the following surface markers: CD69, CD107a, PD-1, CD25, CD62L, CCR7, CD45RO, CD27 and LAG3.

[0741] Changes in assessed surface markers on CAR-expressing CD4+ and CD8+ cells after incubation with Compound 1 and lenalidomide are observed differently in all donors and at different E:T ratios, although expression changes are more pronounced at day 28 compared day 4. On day 4, CD25 and LAG3 are up-regulated for all three donors in response to Compound 1 or lenalidomide treatment, with a greater decrease seen on cells from day 28 compared to day 4. CCR7 tends to decrease in day 28 for all treatment groups, consistent with the possibility that incubation with target cells in the presence of Compound 1 or lenalidomide may shift the T lymphocyte product towards a phenotype associated with terminal differentiated effector status. PD-1 is down-regulated to some extent in all donors and at both E:T ratios on cells at day 4 and day 28 after treatment with Compound 1 or lenalidomide, with more down-regulation occurring on cells at day 28. As shown in Figures 37A and 37B, CD28 expression decreases in a dose-dependent manner in the presence of increasing concentrations of Compound 1 and lenalidomide, respectively, on both CD4+ and CD8+ CAR-expressing T lymphocytes from all three donors. Together, the changes in surface markers at day 28 compared to day 4 are consistent with the ability of Compound 1 and lenalidomide to affect CAR+ T lymphocytes after long-term treatment.

B. Cytolytic function

[0742] On day 24 after the serial stimulation, the cytolytic activity of CAR-expressing anti-CD19 T-lymphocytes was assessed, in general, as described in Example 15B. As shown in FIG. 38, long-term treatment with both Compound 1 and lenalidomide can increase the cytolytic activity of CAR-expressing anti-CD19 T lymphocytes.

C. reproduction

[0743] To evaluate the effect of Compound 1 and lenalidomide on the proliferative potential of CAR+ T lymphocytes, cell numbers of CAR-expressing anti-CD19 T lymphocytes were counted after each stimulation run and cell doublings were calculated.

[0744] As shown in FIG. 39A, at an E:T ratio of 2.5:1, CAR-expressing anti-CD19 T lymphocytes treated with Compound 1 at 500 nM had cell counts comparable to the control treated group, up to 3-4 rounds of stimulation for all donors. Similar results are observed at an E:T ratio of 10:1 for two donors. At a higher Compound concentration of 1,500 nM, the number of doublings of CAR+ cells on subsequent visits is lower than in the untreated control groups. In contrast, after 24 days of Compound 1 treatment at lower concentrations of 10 nM and 100 nM, cell counts of CAR-expressing anti-CD19 T-lymphocytes are higher than for untreated controls from two of the three donors (FIG. 39B).

[0745] As shown in FIG. 40A, at an E:T ratio of 2.5:1 in cells treated with 1000 nM lenalidomide, lower cell doublings are observed in only two donors and not until later stimulation runs. This result shows some activity differences between Compound 1 and lenalidomide, as 500 nM Compound 1 reduces cell counts for all donors at this E:T ratio. With an E:T ratio of 10:1, a decrease in cell doublings is observed after 3-4 stimulation rounds in the presence of 1000 nM lenalidomide, for cells generated from all donors (FIG. 40A). As shown in FIG. 40B, treatment with lenalidomide at a lower concentration of 100 nM increased CAR+ T-lymphocyte counts for two of the three donors.

[0746] The results are consistent with the observation that prolonged treatment of CAR-expressing T-lymphocytes with Compound 1 or lenalidomide at physiologically relevant concentrations may increase the long-term proliferation potential of CAR-expressing T-lymphocytes, while higher concentrations may be detrimental to long-term characteristics

Example 18 Evaluation of the antitumor efficacy of CAR-expressing T-lymphocytes in combination with Compound 1 in vivo

[0747] The antitumor efficacy of CAR-expressing T-lymphocytes in combination with Compound 1 was evaluated by monitoring tumors in a tumor xenograft model. Compositions of CAR-expressing anti-CD19 T lymphocytes containing CD4+ and CD8+ T lymphocytes combined at a ratio of 1:1 were generated essentially as described in Example 1. Compositions of T lymphocytes were generated from three different donors.

[0748] NOD.Cg.Prkdc ^scid IL2rg ^tm1Wjl /SzJ (NSG) mice are intravenously injected with (iv) 0.5 x 10 ⁶ Raji's lymphoma tumor cells (an immortal human B lymphocyte tumor cell line that expresses CD19) that are transfected firefly luciferase (Raji-ffluc). The tumor is allowed to engraft for 6 days and verified using bioluminescent imaging. On Day 7, mice either receive no treatment or receive a single intravenous (iv) injection of anti-CD19 CAR-expressing cells at low dose (0.5 x 10 ⁶ cells) or high dose (1.0 x 10 ⁶ cells) . In one study group (labeled “simultaneous”), mice are treated with 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione (Compound 1) at a dose of 0 .3 mg/kg or vehicle control via intraperitoneal injection one day prior to administration of CAR-expressing cells (Day 6), which continues to be given once a day throughout the study. In the second group (labeled “delayed”), mice are given either vehicle control or 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione (Compound 1) at a dose of 0.3 mg/kg by intraperitoneal injection starting on day 14, which occurs after the peak of proliferation of CAR-expressing T-lymphocytes, and the introduction is continued once a day throughout the study. Tumor mass is assessed by bioluminescence every 10 days. For bioluminescent imaging, mice receive intraperitoneal (ip) injections of luciferin substrate (CaliperLife Sciences, Hopkinton, MA) resuspended in PBS (15 μg/g body weight). Determine the average intensity of radiation (photons/sec/cm ² /stereoradian).

[0749] In this study, combination with Compound 1 was observed to reduce tumor mass and improve survival data, both in the "simultaneous" and "delayed" groups, compared with the introduction of CAR-expressing cells by themselves .

[0750] The present invention is not intended to be limited by the specific embodiments described, which are provided, for example, to illustrate various aspects of the present invention. Various modifications of the described compositions and methods will become apparent from the description and concept of this document. Such variations may be carried out without departing from the true scope and spirit of the present disclosure and are intended to fall within the scope of the present disclosure.

Sequence list

SEQID NO. SUBSEQUENCE DESCRIPTION one ESKYGPPCPPCP Spacer (IgG4 hinge) (aa)
Homo sapiens 2 GAATCTAAGTACGGACCGCCCTGCCCCCCTTGCCCT Spacer (IgG4 hinge) (nt)
Homo sapiens 3 ESKYGPPCPPCPGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Hinge spacer CH3
Homo sapiens four ESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Hinge spacer CH2-CH3
Homo sapiens 5 RWPESPKAQASSVPTAQPQAEGSLAKATTAPATTRNTGRGGEEKKKEKEKEEQEERETKTPECPSHTQPLGVYLLTPAVQDLWLRDKATFTCFVVGSDLKDAHLTWEVAGKVPTGGVEEGLLERHSNGSQSQHSRLTLPRSL
WNAGTSVTCTLNHPSLPPQRLMALREPAAQAPVKLSLNLLASSDPPEAASWLLCEVSGFSPPNILLMWLEDQREVNTSGFAPARPPPQPGSTTFWAWSVLRVPAPPSPQPATYTCVVSHEDSRTLLNASRSLEVSYVTDH IgD-hinge-Fc
Homo sapiens 6 LEGGGEGRGSLTCGDVEENPGPR Faux T2A 7 RKVCNGIGIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVKEITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISDGDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWGPEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGPDNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEGCPTNGPKIPSIATGMVGALLLLLVVALGIGLFM Artificial tEGFR eight FWVLVVVGGVLACYSLLVTVAFIFWV CD28 (amino acids 153-179, Accession No. P10747)
Homo sapiens 9 IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP FWVLVVVGGVLACYSLLVTVAFIIFWV CD28 (amino acids 114-179, Accession No. P10747)
Homo sapiens ten RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS CD28 (amino acids 180-220 from P10747)
Homo sapiens eleven RSKRSRGGHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS CD28 (LL - GG)
Homo sapiens 12 KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL 4-1BB (amino acids 214-255 from Q07011.1)
Homo sapiens 13 RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYN ELQKDKMAEA YSEIGMKGER RRGKGHDGLY QGLSTATKDTYDALHMQALP PR CD3 zeta
Homo sapiens fourteen RVKFSRSAEPPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYN ELQKDKMAEA YSEIGMKGER RRGKGHDGLY QGLSTATKDTYDALHMQALP PR CD3 zeta
Homo sapiens fifteen RVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYN ELQKDKMAEA YSEIGMKGER RRGKGHDGLY QGLSTATKDTYDALHMQALP PR CD3 zeta
Homo sapiens 16 PGGG-(SGGGG)5-P- where P is proline, G is glycine and S is serine Linker 17 GSADDAKKDAAKKDGKS Linker eighteen MLQMAGQCSQNEYFDSLLHACIPCQLRCSSNTPPLTCQRYCNASVTNSVKGTNA Human BCMA extracellular domain (GenBank No. NP_001183.2) 19 GGGGS Linker sequence twenty PKSSDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDQFFLYSKLTVDKW Fc modified human IgG1 21 MPLLLLPLLWAGALA Signal peptide CD33 22 MPLLLLLPLLWAGALAMLQMAGQCSQNEYFDSLLHACIPCQLRCSSNTPPLTCQRYCNASVTNSVKGTNAGGGGSPKSSDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK BCMA-Fc construction 23 EGRGSLLTCGDVEENPGP T2A 24 GSGATNFSLLKQAGDVEENPGP P2A 25 ATNFSLLKQAGDVEENPGP P2A 26 QCTNYALLKLAGDVESNPGP E2A 27 VKQTLNFDLLKLAGDVESNPGP F2A 28 RKVCNGIGIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVKEITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISDGDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWGPEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGPDNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEGCPTNGPKIPSIATGMVGALLLLLVVALGIGLFM Artificial
tEGFR 29 ESKYGPPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSLVK Hinge spacer CH2-CH3
Homo sapiens thirty QIQLVQSGPELKKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPAYAYDFRGRFAFSLETSASTAYLQINNLKYEDTATYFCALDYSYAMDYWGQGTSVTVSS Variable heavy (V _H ) anti-BCMA 31 DIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQKPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIK Variable light (VL) anti-BCMA 32 QIQLVQSGPDLKKPGETVKLSCKASGYTFFTNFGMNWVKQAPGKGFKWMAWINTYTGESYFADDFKGRFAFSVETSATTAYLQINNLKTEDTATYFCARGEIYYGYDGGFAYWGQGTLVTVSA Variable heavy (V _H ) anti-BCMA 33 DVVMTQSHRFMSTSVGDRVSITCRASQDVNTAVSWYQQKPGQSPKLLIFSASYRYTGVPDRFTGSGSGADFTLTISSVQAEDLAVYYCQQHYSTPWTFGGGTKLDIK Variable light (VL) anti-BCMA 34 EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGHVTISADKSISTAYLQWSSLKASDTAMYYCARYSGSFDNWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 35 SYELTQPPSASGTPGQRVTMSCSGTSSNIGSHSVNWYQQLPGTAPKLLIYTNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDGSLNGLVFGGGTKLTVLG Variable light (VL) anti-BCMA 36 EVQLVQSGAEMKKPGASLKLSCKASGYTFIDYYVYWMRQAPGQGLESMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAMYYCARSQRDGYMDYWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 37 QSALTQPASVSASPGQSIAISCTGTSSDVGWYQQHPGKAPKLMIYEDSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSNTRSSTLVFGGGTKLTVLG Variable light (VL) anti-BCMA 38 EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGIANYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCARSGYSKSIVSYMDYWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 39 LPVLTQPPSTSGTPGQRVTVSCSGSSSNIGSNVVFWYQQLPGTAPKLVIYRNNQRPSGVPDRFSVSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGYVFGTGTKVTVLG Variable light (VL) anti-BCMA 40 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSGYGSYRWEDSWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 41 QAVLTQPPSASGTPGQRVTISCSGSSSNIGSNYVFWYQQLPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSASYVFGTGTKVTVLG Variable light (VL) anti-BCMA 42 QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYYMHWVRQAPGQRLEWMGWINPNSGGTNYAQKFQDRITVTRDTSSNTGYMELTRLRSDDTAVYYCARSPYSGVLDKWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 43 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGFDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGYVFGTGTKVTVLG Variable Light (VL) Anti-BCMA 44 DYGVS FMC63 CDR H1 45 VIWGSETTYYNSALKS FMC63 CDR H2 46 YAMDYWG FMC63 CDR H3 47 HYYYGGSYAMDY FMC63HC-CDR3 48 RASQDISKYLN FMC63 CDR L1 49 SRLHSGV FMC63 CDR L2 fifty HTSRLHS FMC63 LC-CDR2 51 GNTLPYTFG FMC63 CDR L3 52 QQGNTLPYT FMC63 LC-CDR3 53 EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS FMC63 V _H 54 DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT FMC63 VL 55 DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGSTSGSGKPGSGEGSTKGEVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS FMC63 scFv 56 KASQNVGTNVA SJ25C1 CDR L1 57 SATYRNS SJ25C1 CDR L2 58 QQYNRYPYT SJ25C1 CDR L3 59 SYWMN SJ25C1 CDR H1 60 QIYPGDGDTNYNGKFKG SJ25C1 CDR H2 61 KTISSVVDFYFDY SJ25C1 CDR H3 62 EVKLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIYPGDGDTNYNGKFKGQATLTADKSSSTAYMQLSGLTSEDSAVYFCARKTISSVVDFYFDYWGQGTTVTVSS SJ25C1 V _H 63 DIELTQSPKFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSPKPLIYSATYRNSGVPDRFTGSGSGTDFTLTITNVQSKDLADYFCQQYNRYPYTSGGGTKLEIKR SJ25C1 VL 64 GGGGSGGGGSGGGGS Linker 65 EVKLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIYPGDGDTNYNGKFKGQATLTADKSSSTAYMQLSGLTSEDSAVYFCARKTISSVVDFYFDYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPKFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSPKPLIYSATYRNSGVPDRFTGSGSGTDFTLTITNVQSKDLADYFCQQYNRYPYTSGGGTKLEIKR SJ25C1 scFv 66 HYYYGGSYAMDY FMC63HC-CDR3 67 HTSRLHS FMC63 LC-CDR2 68 QQGNTLPYT FMC63 LC-CDR3 69 gacatccagatgacccagaccacctccagcctgagcgccagcctgggcgaccgggtgaccatcagctgccgggccagccaggacatcagcaagtacctgaactggtatcagcagaagcccgacggcaccgtcaagctgctgatctaccacaccagccggctgcacagcggcgtgcccagccggtttagcggcagcggctccggcaccgactacagcctgaccatctccaacctggaacaggaagatatcgccacctacttttgccagcagggcaacacactgccctacacctttggcggcggaacaaagctggaaatcaccggcagcacctccggcagcggcaagcctggcagcggcgagggcagcaccaagggcgaggtgaagctgcaggaaagcggccctggcctggtggcccccagccagagcctgagcgtgacctgcaccgtgagcggcgtgagcctgcccgactacggcgtgagctggatccggcagccccccaggaagggcctggaatggctgggcgtgatctggggcagcgagaccacctactacaacagcgccctgaagagccggctgaccatcatcaaggacaacagcaagagccaggtgttcctgaagatgaacagcctgcagaccgacgacaccgccatctactactgcgccaagcactactactacggcggcagctacgccatggactactggggccagggcaccagcgtgaccgtgagcagc scFv encoding sequence 70 GSTSGSGKPGSGEGSTKG Linker 71 GGGS Linker 72 GGGGSGGGGSGGGGS Linker 73 GSTSGSGKPGSGEGSTKG Linker 74 SRGGGGSGGGGSGGGGSLEMA Linker 75 MALPVTALLLPLALLLHAARP Signal peptide CD8a 76 METDTLLLWVLLLWVPGSTG Signal peptide 77 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARAEMGAVFDIWGQGTMVTVSS Variable heavy (V _H ) anti-BCMA 78 EIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRISWPFTFGGGTKVEIK Variable light (VL) anti-BCMA 79 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGTYLGGLWYFDLWGRGTLVTVSS Variable heavy (V _H ) anti-BCMA 80 DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGLGLPLTFGGGTKVEIK Variable light (VL) anti-BCMA 81 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPGGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARESWPMDVWGQGTTVTVSS Variable heavy (V _H ) anti-BCMA 82 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYAAYPTFGGGTKVEIK Variable light (VL) anti-BCMA 83 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPPGKGLEWIGSISYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGRGYATSLAFDIWGQGTMVTVSS Variable heavy (V _H ) anti-BCMA 84 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRHVWPPTFGGGTKVEIK Variable light (VL) anti-BCMA 85 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSTISSSSSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARGSQEHLIFDYWGQGTLVTVSS Variable severe (V _H ) Anti-BCMA 86 EIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRFYYPWTFGGGTKVEIK Variable Light (VL) Anti-BCMA 87 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTDFWSGSPPGLDYWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 88 DIQLTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYGASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQIYTFPFTFGGGTKVEIK Variable light (VL) anti-BCMA 89 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARTPEYSSSIWHYYYGMDVWGQGTTVTVSS Variable heavy (V _H ) anti-BCMA 90 DIVMTQSPDSLAVSLGERATINCKSSQSVLYSSNNKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQFAHTPFTFGGGTKVEIK Variable light (VL) anti-BCMA 91 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVKGPLQEPPYDYGMDVWGQGTTVTVSS Variable heavy (V _H ) anti-BCMA 92 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYSASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQHHVWPLTFGGGTKVEIK Variable light (VL) anti-BCMA 93 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGIANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGGYYSHDMWSEDWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 94 LPVLTQPPSASGTPGQRVTISCSGRSSNIGSNSVNWYRQLPGAAPKLLIYSNNQRPPGVPVRFSGSKSGTSASLAISGLQSEDEATYYCATWDDNLNVHYVFGTGTKVTVLG Variable light (VL) anti-BCMA 95 QVQLVQSGSELKKPGASVKVSCKASGYTFTDYSINWVRQAPGQGLEWMGWINTETREPAYAYDFRGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCARDYSYAMDYWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 96 DIVLTQSPASLAVSLGERATINCRASESVSVIGAHLIHWYQQKPGQPPKLLIYLASNLETGVPARFSGSGSGTDFTLTISSLQAEDAAIYYCLQSRIFPRTFGQGTKLEIK Variable light (VL) anti-BCMA 97 EVQLVESGGGLVQPGGSLRLSCAVSGFALSNHGMSWVRRAPGKGLEWVSGIVYSGSTYYAASVKGRFTISRDNSRNTLYLQMNSLRPEDTAIYYCSAHGGESDVWGQGTTVTVSS Variable heavy (V _H ) anti-BCMA 98 DIQLTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKVEIK Variable light (VL) anti-BCMA 99 QVQLVESGGGLVQPGRSLRLSCAASGFTFSNYAMSWVRQAPGKGLGWVSGISRSGENTYYADSVKGRFTISRDNSKNTLYLQMNSLRDEDTAVYYCARSPAHYYGGMDVWGQGTTVTVSS Variable heavy (V _H ) anti-BCMA 100 DIVLTQSPGTLSLSPGERATLSCRASQSISSSFLAWYQQKPGQAPRLLIYGASRRATGIPDRFSGSGSGTDFTLTISRLEPEDSAVYCQQYHSSPSWTFGQGTKLEIK Variable light (VL) anti-BCMA 101 QVQLVESGGGLVQPGGSLRLSCAVSGFALSNHGMSWVRRAPGKGLEWVSGIVYSGSTYYAASVKGRFTISRDNSRNTLYLQMNSLRPEDTAIYYCSAHGGESDVWGQGTTVTVSS Variable heavy (V _H ) anti-BCMA 102 DIRLTQSPSPLSASVGDRVTITCQASEDINKFLNWYHQTPGKAPKLLIYDASTLQTGVPSRFSGSGSGTDFTLTINSLQPEDIGTYYCQQYESLPLTFGGGTKVEIK Variable light (VL) anti-BCMA 103 EVQLVESGGGLVQPGGSLRLSCAVSGFALSNHGMSWVRRAPGKGLEWVSGIVYSGSTYYAASVKGRFTISRDNSRNTLYLQMNSLRPEDTAIYYCSAHGGESDVWGQGTTVTVSS Variable heavy (V _H ) anti-BCMA 104 EIVLTQSPGTLSLSPGERATLSCRASQSIGSSSLAWYQQKPGQAPRLLMYGASSRASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYCQQYAGSPPFTFGQGTKVEIK Variable light (VL) anti-BCMA 105 QIQLVQSGPELKKPGETVKISCKASGYTFRHYSMNWVKQAPGKGLKWMGRINTESGVPIYADDFKGRFAFSVETSASTAYLVINNLKDEDTASYFCSNDYLYSLDFWGQGTALTVSS Variable heavy (V _H ) anti-BCMA 106 DIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIYWYQQKPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIK Variable light (VL) anti-BCMA 107 QIQLVQSGPELKKPGETVKISCKASGYTFTHYSMNWVKQAPGKGLKWMGRINTETGEPLYADDFKGRFAFSLETSASTAYLVINNLKNEDTATFFCSNDYLYSCDYWGQGTTLTVSS Variable heavy (V _H ) anti-BCMA 108 DIVLTQSPASLAMSLGKRATISCRASESVSVIGAHLIHWYQQKPGQPPKLLIYLASNLETGVPARFSGSGSGTDFTLTIDPVEEDDVAIYSCLQSRIFPRTFGGGTKLEIK Variable light (VL) anti-BCMA 109 QVQLVQSGAEVKKPGASVKVSCKASGYSFPDYYINWVRQAPGQGLEWMGWIYFASGNSEYNQKFTGRVTMTRDTSINTAYMELSSLTSEDTAVYFCASLYDYDWYFDVWGQGTMVTVSS Variable heavy (V _H ) anti-BCMA 110 DIVMTQTPLSLSVTPGQPASISCKSSQSLVHSNGNTYLHWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGIYYCSQSSIYPWTFGQGTKLEIK Variable light (VL) anti-BCMA 111 QVQLVQSGAEVKKPGASVKVSCKASGYSFPDYYINWVRQAPGQGLEWMGWIYFASGNSEYNQKFTGRVTMTRDTSSSTAYMELSSLRSEDTAVYFCASLYDYDWYFDVWGQGTMVTVSS Variable heavy (V _H ) anti-BCMA 112 DIVMTQTPLSLSVTPGEPASISCKSSQSLVHSNGNTYLHWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGADFTLKISRVEAEDVGVYYCAETSHVPWTFGQGTKLEIK Variable light (VL) anti-BCMA 113 QVQLVESGGGLVQPGGSLRLSCEASGFTLDYYAIGWFRQAPGKEREGVICISRSDGSTYYADSVKGRFTISRDNAKKTVYLQMISLKPEDTAAYYCAAGADCSGYLRDYEFRGQGTQVTVSS sdAb anti-BCMA 114 IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP Spacer CD28 115 IYIWAPLAGTCGVLLLSLVITLYCN CD8aTM 116 LDNEKSNGTIIHVKGKHLCPSPLFPGPSKP Spacer CD28 (truncated) 117 PTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD Hinge CD8a 118 TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD Hinge CD8a 119 FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD Hinge CD8a 120 DTGLYICKVELMYPPPYYLGIGNGTQIYVIDPEPCPDSD Hinge CTLA4 121 FLLWILAAVSSGLFFYSFLLTAVS CTLA4TM 122 QIKESLRAELRVTERRAEVPTAHPSSPSPRPAGQFQTLV Hinge PD-1 123 VGVVGGLLGSLVLLVWVLAVI PD-1TM 124 GLAVSTISSFFPPGYQ Hinge FcγRIIIa 125 EPKSPDKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMIARTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDQFFLYNKLSVDKW IgG1 hinge 126 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARAEMGAVFDIWGQGTMVTVSSGSTSGSGKPGSGEGSTKGEIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRISWPFTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 127 EIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRISWPFTFGGGTKVEIKRGSTSGSGKPGSGEGSTKGEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARAEMGAVFDIWGQGTMVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 128 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGTYLGGLWYFDLWGRGTLVTVSSGSTSGSGKPGSGEGSTKGDIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGLGLPLTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 129 DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGLGLPLTFGGGTKVEIKRGSTSGSGKPGSGEGSTKGQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGTYLGGLWYFDLWGRGTLVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 130 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPGGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARESWPMDVWGQGTTVTVSSGSTSGSGKPGSGEGSTKGEIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYAAYPTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 131 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYAAYPTFGGGTKVEIKRGSTSGSGKPGSGEGSTKGQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPGGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARESWPMDVWGQGTTVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 132 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPPGKGLEWIGSISYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGRGYATSLAFDIWGQGTMVTVSSGSTSGSGKPGSGEGSTKGEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRHVWPPTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 133 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRHVWPPTFGGGTKVEIKRGSTSGSGKPGSGEGSTKGQLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPPGKGLEWIGSISYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGRGYATSLAFDIWGQGTMVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 134 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSTISSSSSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARGSQEHLIFDYWGQGTLVTVSSGSTSGSGKPGSGEGSTKGEIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRFYYPWTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 135 EIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRFYYPWTFGGGTKVEIKRGSTSGSGKPGSGEGSTKGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSTISSSSSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARGSQEHLIFDYWGQGTLVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 136 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTDFWSGSPPGLDYWGQGTLVTVSSGSTSGSGKPGSGEGSTKGDIQLTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYGASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQIYTFPFTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 137 DIQLTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYGASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQIYTFPFTFGGGTKVEIKRGSTSGSGKPGSGEGSTKGQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTDFWSGSPPGLDYWGQGTLVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 138 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARTPEYSSSIWHYYYGMDVWGQGTTVTVSSGSTSGSGKPGSGEGSTKGDIVMTQSPDSLAVSLGERATINCKSSQSVLYSSNNKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQFAHTPFTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 139 DIVMTQSPDSLAVSLGERATINCKSSQSVLYSSNNKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQFAHTPFTFGGGTKVEIKRGSTSGSGKPGSGEGSTKGQVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARTPEYSSSIWHYYYGMDVWGQGTTVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 140 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVKGPLQEPPYDYGMDVWGQGTTVTVSSGSTSGSGKPGSGEGSTKGEIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYSASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQHHVWPLTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 141 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYSASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQHHVWPLTFGGGTKVEIKRGSTSGSGKPGSGEGSTKGQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVKGPLQEPPYDYGMDVWGQGTTVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 142 QSALTQPASVSASPGQSIAISCTGTSSDVGWYQQHPGKAPKLMIYEDSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSNTRSSTLVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEMKKPGASLKLSCKASGYTFIDYYVYWMRQAPGQGLESMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAMYYCARSQRDGYMDYWGQGTLVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 143 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGFDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGASVKVSCKASGYTFTDYYMHWVRQAPGQRLEWMGWINPNSGGTNYAQKFQDRITVTRDTSSNTGYMELTRLRSDDTAVYYCARSPYSGVLDKWGQGTLVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 144 SYELTQPPSASGTPGQRVTMSCSGTSSNIGSHSVNWYQQLPGTAPKLLIYTNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDGSLNGLVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGHVTISADKSISTAYLQWSSLKASDTAMYYCARYSGSFDNWGQGTLVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 145 LPVLTQPPSASGTPGQRVTISCSGRSSNIGSNSVNWYRQLPGAAPKLLIYSNNQRPPGVPVRFSGSKSGTSASLAISGLQSEDEATYYCATWDDNLNVHYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGIANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGGYYSHDMWSEDWGQGTLVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 146 QAVLTQPPSASGTPGQRVTISCSGSSSNIGSNYVFWYQQLPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSASYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSGYGSYRWEDSWGQGTLVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 147 LPVLTQPPSASGTPGQRVTISCSGRSSNIGSNSVNWYRQLPGAAPKLLIYSNNQRPPGVPVRFSGSKSGTSASLAISGLQSEDEATYYCATWDDNLNVHYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGIANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGGYYSHDMWSEDWGQGTLVTVSSAAAPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSAEPPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 148 SYELTQPPSASGTPGQRVTMSCSGTSSNIGSHSVNWYQQLPGTAPKLLIYTNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDGSLNGLVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGHVTISADKSISTAYLQWSSLKASDTAMYYCARYSGSFDNWGQGTLVTVSSAAAPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSAEPPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 149 QAVLTQPPSASGTPGQRVTISCSGSSSNIGSNYVFWYQQLPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSASYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSGYGSYRWEDSWGQGTLVTVSSAAAPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSAEPPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 150 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGFDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGASVKVSCKASGYTFTDYYMHWVRQAPGQRLEWMGWINPNSGGTNYAQKFQDRITVTRDTSSNTGYMELTRLRSDDTAVYYCARSPYSGVLDKWGQGTLVTVSSAAAPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSAEPPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 151 QSALTQPASVSASPGQSIAISCTGTSSDVGWYQQHPGKAPKLMIYEDSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSNTRSSTLVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEMKKPGASLKLSCKASGYTFIDYYVYWMRQAPGQGLESMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAMYYCARSQRDGYMDYWGQGTLVTVSSAAAPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSAEPPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 152 DIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQKPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIKGSTSGSGKPGSGEGSTKGQIQLVQSGPELKKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPAYAYDFRGRFAFSLETSASTAYLQINNLKYEDTATYFCALDYSYAMDYWGQGTSVTVSSAAATTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 153 DIVLTQSPASLAVSLGERATINCRASESVSVIGAHLIHWYQQKPGQPPKLLIYLASNLETGVPARFSGSGSGTDFTLTISSLQAEDAAIYYCLQSRIFPRTFGQGTKLEIKGSTSGSGKPGSGEGSTKGQVQLVQSGSELKKPGASVKVSCKASGYTFTDYSINWVRQAPGQGLEWMGWINTETREPAYAYDFRGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCARDYSYAMDYWGQGTLVTVSSAAATTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 154 DIVLTQSPASLAVSLGERATINCRASESVSVIGAHLIHWYQQKPGQPPKLLIYLASNLETGVPARFSGSGSGTDFTLTISSLQAEDAAIYYCLQSRIFPRTFGQGTKLEIKGSTSGSGKPGSGEGSTKGQVQLVQSGSELKKPGASVKVSCKASGYTFTDYSINWVRQAPGQGLEWMGWINTETREPAYAYDFRGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCARDYSYAMDYWGQGTLVTVSSAAADTGLYICKVELMYPPPYYLGIGNGTQIYVIDPEPCPDSDFLLWILAAVSSGLFFYSFLLTAVSKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 155 DIVLTQSPASLAVSLGERATINCRASESVSVIGAHLIHWYQQKPGQPPKLLIYLASNLETGVPARFSGSGSGTDFTLTISSLQAEDAAIYYCLQSRIFPRTFGQGTKLEIKGSTSGSGKPGSGEGSTKGQVQLVQSGSELKKPGASVKVSCKASGYTFTDYSINWVRQAPGQGLEWMGWINTETREPAYAYDFRGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCARDYSYAMDYWGQGTLVTVSSAAAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVLAVICSKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 156 EVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWIRQAPGKGLEWVSYISSSGSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVDGDYTEDYWGQGTLVTVSSGGGGSGGGGSGGGGSQSALTQPASVSGSPGQSITISCTGSSSDVGKYNLVSWYQQPPGKAPKLIIYDVNKRPSGVSNRFSGSKSGNTATLTISGLQGDDEADYYCSSYGGSRSYVFGTGTKVTVLESKYGPPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKMFWVLVVVGGVLACYSLLVTVAFIIFWVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 157 EVQLVQSGGGLVQPGRSLRLSCTASGFTFGDYAMSWFKQAPGKGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLKTEDTAVYYCAAWSAPTDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPAFLSASVGDRVTVTCRASQGISNYLAWYQQKPGNAPRLLIYSASTLQSGVPSRFRGTGYGTEFSLTIDSLQPEDFATYYCQQSYTSRQTFGPGTRLDIKESKYGPPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKMFWVLVVVGGVLACYSLLVTVAFIIFWVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 158 EVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWIRQAPGKGLEWVSYISSSGSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVDGPPSFDIWGQGTMVTVSSGGGGSGGGGSGGGGSSYVLTQPPSVSVAPGQTARITCGANNIGSKSVHWYQQKPGQAPMLVVYDDDDRPSGIPERFSGSNSGNTATLTISGVEAGDEADYFCHLWDRSRDHYVFGTGTKLTVLESKYGPPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKMFWVLVVVGGVLACYSLLVTVAFIIFWVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 159 SYELTQPPSASGTPGQRVTMSCSGTSSNIGSHSVNWYQQLPGTAPKLLIYTNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDGSLNGLVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGHVTISADKSISTAYLQWSSLKASDTAMYYCARYSGSFDNWGQGTLVTVSSESKYGPPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKMFWVLVVVGGVLACYSLLVTVAFIIFWVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 160 QSALTQPASVSASPGQSIAISCTGTSSDVGWYQQHPGKAPKLMIYEDSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSNTRSSTLVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEMKKPGASLKLSCKASGYTFIDYYVYWMRQAPGQGLESMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAMYYCARSQRDGYMDYWGQGTLVTVSSESKYGPPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKMFWVLVVVGGVLACYSLLVTVAFIIFWVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 161 QSALTQPASVSASPGQSIAISCTGTSSDVGWYQQHPGKAPKLMIYEDSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSNTRSSTLVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEMKKPGASLKLSCKASGYTFIDYYVYWMRQAPGQGLESMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAMYYCARSQRDGYMDYWGQGTLVTVSSESKYGPPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKMFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 162 EVQLVESGGGLVQPGGSLRLSCAVSGFALSNHGMSWVRRAPGKGLEWVSGIVYSGSTYYAASVKGRFTISRDNSRNTLYLQMNSLRPEDTAIYYCSAHGGESDVWGQGTTVTVSSASGGGGSGGRASGGGGSDIQLTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKVEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 163 QVQLVESGGGLVQPGRSLRLSCAASGFTFSNYAMSWVRQAPGKGLGWVSGISRSGENTYYADSVKGRFTISRDNSKNTLYLQMNSLRDEDTAVYYCARSPAHYYGGMDVWGQGTTVTVSSASGGGGSGGRASGGGGSDIVLTQSPGTLSLSPGERATLSCRASQSISSSFLAWYQQKPGQAPRLLIYGASRRATGIPDRFSGSGSGTDFTLTISRLEPEDSAVYYCQQYHSSPSWTFGQGTKLEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 164 QVQLVESGGGLVQPGGSLRLSCAVSGFALSNHGMSWVRRAPGKGLEWVSGIVYSGSTYYAASVKGRFTISRDNSRNTLYLQMNSLRPEDTAIYYCSAHGGESDVWGQGTTVTVSSASGGGGSGGRASGGGGSDIRLTQSPSPLSASVGDRVTITCQASEDINKFLNWYHQTPGKAPKLLIYDASTLQTGVPSRFSGSGSGTDFTLTINSLQPEDIGTYYCQQYESLPLTFGGGTKVEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 165 EVQLVESGGGLVQPGGSLRLSCAVSGFALSNHGMSWVRRAPGKGLEWVSGIVYSGSTYYAASVKGRFTISRDNSRNTLYLQMNSLRPEDTAIYYCSAHGGESDVWGQGTTVTVSSASGGGGSGGRASGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSIGSSSLAWYQQKPGQAPRLLMYGASSRASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYAGSPPFTFGQGTKVEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 166 QIQLVQSGPDLKKPGETVKLSCKASGYTFTNFGMNWVKQAPGKGFKWMAWINTYTGESYFADDFKGRFAFSVETSATTAYLQINNLKTEDTATYFCARGEIYYGYDGGFAYWGQGTLVTVSAGGGGSGGGGSGGGGSDVVMTQSHRFMSTSVGDRVSITCRASQDVNTAVSWYQQKPGQSPKLLIFSASYRYTGVPDRFTGSGSGADFTLTISSVQAEDLAVYYCQQHYSTPWTFGGGTKLDIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 167 QIQLVQSGPELKKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPAYAYDFRGRFAFSLETSASTAYLQINNLKYEDTATYFCALDYSYAMDYWGQGTSVTVSSGGGGSGGGGSGGGGSQIQLVQSGPELKKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPAYAYDFRGRFAFSLETSASTAYLQINNLKYEDTATYFCALDYSYAMDYWGQGTSVTVSSTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 168 QIQLVQSGPELKKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPAYAYDFRGRFAFSLETSASTAYLQINNLKYEDTATYFCALDYSYAMDYWGQGTSVTVSSGGGGSGGGGSGGGGSDIVLTQSPASLAMSLGKRATISCRASESVSVIGAHLIHWYQQKPGQPPKLLIYLASNLETGVPARFSGSGSGTDFTLTIDPVEEDDVAIYSCLQSRIFPRTFGGGTKLEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 169 QIQLVQSGPELKKPGETVKISCKASGYTFRHYSMNWVKQAPGKGLKWMGRINTESGVPIYADDFKGRFAFSVETSASTAYLVINNLKDEDTASYFCSNDYLYSLDFWGQGTALTVSSGGGGSGGGGSGGGGSDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIYWYQQKPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 170 QIQLVQSGPELKKPGETVKISCKASGYTFTHYSMNWVKQAPGKGLKWMGRINTETGEPLYADDFKGRFAFSLETSASTAYLVINNLKNEDTATFFCSNDYLYSCDYWGQGTTLTVSSGGGGSGGGGSGGGGSDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIYWYQQKPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 171 DIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQKPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIKGSTSGSGKPGSGEGSTKGQIQLVQSGPELKKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPAYAYDFRGRFAFSLETSASTAYLQINNLKYEDTATYFCALDYSYAMDYWGQGTSVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 172 QVQLVQSGAEVKKPGASVKVSCKASGYSFPDYYINWVRQAPGQGLEWMGWIYFASGNSEYNQKFTGRVTMTRDTSINTAYMELSSLTSEDTAVYFCASLYDYDWYFDVWGQGTMVTVSSGGGGSGGGGSGGGGSDIVMTQTPLSLSVTPGQPASISCKSSQSLVHSNGNTYLHWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGIYYCSQSSIYPWTFGQGTKLEIKGLAVSTISSFFPPGYQIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 173 QVQLVQSGAEVKKPGASVKVSCKASGYSFPDYYINWVRQAPGQGLEWMGWIYFASGNSEYNQKFTGRVTMTRDTSINTAYMELSSLTSEDTAVYFCASLYDYDWYFDVWGQGTMVTVSSGGGGSGGGGSGGGGSDIVMTQTPLSLSVTPGQPASISCKSSQSLVHSNGNTYLHWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGIYYCSQSSIYPWTFGQGTKLEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 174 QVQLVQSGAEVKKPGASVKVSCKASGYSFPDYYINWVRQAPGQGLEWMGWIYFASGNSEYNQKFTGRVTMTRDTSINTAYMELSSLTSEDTAVYFCASLYDYDWYFDVWGQGTMVTVSSGGGGSGGGGSGGGGSDIVMTQTPLSLSVTPGQPASISCKSSQSLVHSNGNTYLHWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGIYYCSQSSIYPWTFGQGTKLEIKEPKSPDKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMIARTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 175 QVQLVQSGAEVKKPGASVKVSCKASGYSFPDYYINWVRQAPGQGLEWMGWIYFASGNSEYNQKFTGRVTMTRDTSSSTAYMELSSLRSEDTAVYFCASLYDYDWYFDVWGQGTMVTVSSGGGGSGGGGSGGGGSDIVMTQTPLSLSVTPGEPASISCKSSQSLVHSNGNTYLHWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGADFTLKISRVEAEDVGVYYCAETSHVPWTFGQGTKLEIKGLAVSTISSFFPPGYQIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 176 QVQLVQSGAEVKKPGASVKVSCKASGYSFPDYYINWVRQAPGQGLEWMGWIYFASGNSEYNQKFTGRVTMTRDTSSSTAYMELSSLRSEDTAVYFCASLYDYDWYFDVWGQGTMVTVSSGGGGSGGGGSGGGGSDIVMTQTPLSLSVTPGEPASISCKSSQSLVHSNGNTYLHWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGADFTLKISRVEAEDVGVYYCAETSHVPWTFGQGTKLEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 177 QVQLVQSGAEVKKPGASVKVSCKASGYSFPDYYINWVRQAPGQGLEWMGWIYFASGNSEYNQKFTGRVTMTRDTSSSTAYMELSSLRSEDTAVYFCASLYDYDWYFDVWGQGTMVTVSSGGGGSGGGGSGGGGSDIVMTQTPLSLSVTPGEPASISCKSSQSLVHSNGNTYLHWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGADFTLKISRVEAEDVGVYYCAETSHVPWTFGQGTKLEIKEPKSPDKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMIARTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CAR anti-BCMA 178 IYIWAPLAGTCGVLLLSLVITLYCNHRN CD8aTM 179 IYIWAPLAGTCGVLLLSLVIT CD8aTM 180 RAAA Peptide binding 181 EVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWIRQAPGKGLEWVSYISSSGSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVDGDYTEDYWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 182 QSALTQPASVSGSPGQSITISCTGSSSDVGKYNLVSWYQQPPGKAPKLIIYDVNKRPSGVSNRFSGSKSGNTATLTISGLQGDDEADYYCSSYGGSRSYVFGTGTKVTVL Variable light (VL) anti-BCMA 183 EVQLVQSGGGLVQPGRSLRLSCTASGFTFGDYAMSWFRQAPGKGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLKTEDTAVYYCAAWSAPTDYWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 184 DIQMTQSPAFLSASVGDRVTVTCRASQGISNYLAWYQQKPGNAPRLLIYSASTLQSGVPSRFRGTGYGTEFSLTIDSLQPEDFATYYCQQSYTSRQTFGPGTRLDIK Variable light (VL) anti-BCMA 185 EVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWIRQAPGKGLEWVSYISSSGSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVDGPPSFDIWGQGTMVTVSS Variable heavy (V _H ) anti-BCMA 186 SYVLTQPPSVSVAPGQTARITCGANNIGSKSVHWYQQKPGQAPMLVVYDDDDRPSGIPERFSGSNSGNTATLTISGVEAGDEADYFCHLWDRSRDHYVFGTGTKLTVL Variable light (VL) anti-BCMA 187 EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGIANYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCARSGYSKSIVSYMDYWGQGTLVTVSS Variable heavy (V _H ) anti-BCMA 188 LPVLTQPPSTSGTPGQRVTVSCSGSSSNIGSNVVFWYQQLPGTAPKLVIYRNNQRPSGVPDRFSVSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGYVFGTGTKVTVLG Variable light (VL) anti-BCMA 189 ASGGGGSGGRASGGGGS Linker

--->

SEQUENCE LIST

<110> JUNO THERAPEUTICS, INC.

PORTS, Michael

WORKS, Melissa

<120> COMBINATION OF CELL THERAPY AND IMMUNOMODULATOR COMPOUND

<130> 735042009640

<140> not assigned yet

<141> at the same time as this

<150> 62/492.947

<151> 2017-05-01

<150> 62/538.670

<151> 2017-07-29

<150> 62/549.390

<151> 2017-08-23

<150> 62/580.433

<151> 2017-11-01

<150> 62/596.753

<151> 2017-12-08

<160> 189

<170> FastSEQ for Windows Version 4.0

<210> 1

<211> 12

<212> PRT

<213> Homo sapiens

<220>

<223> Spacer (IgG4 Hinge)

<400> 1

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro

1 5 10

<210> 2

<211> 36

<212> DNA

<213> Homo sapiens

<220>

<223> Spacer (IgG4 Hinge)

<400> 2

gaatctaagt acggaccgcc ctgcccccct tgccct 36

<210> 3

<211> 119

<212> PRT

<213> Homo sapiens

<220>

<223> Spacer Hinge-CH3

<400> 3

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Gly Gln Pro Arg

1 5 10 15

Glu Pro Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr Lys

20 25 30

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

35 40 45

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

50 55 60

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

65 70 75 80

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

85 90 95

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

100 105 110

Leu Ser Leu Ser Leu Gly Lys

115

<210> 4

<211> 229

<212> PRT

<213> Homo sapiens

<220>

<223> Spacer Hinge-CH2-CH3

<400> 4

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe

1 5 10 15

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

20 25 30

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

35 40 45

Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val

50 55 60

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser

65 70 75 80

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

85 90 95

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser

100 105 110

Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

115 120 125

Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln

130 135 140

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

145 150 155 160

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

165 170 175

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu

180 185 190

Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser

195 200 205

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

210 215 220

Leu Ser Leu Gly Lys

225

<210> 5

<211> 282

<212> PRT

<213> Homo sapiens

<220>

<223> IgD-Hinge-Fc

<400> 5

Arg Trp Pro Glu Ser Pro Lys Ala Gln Ala Ser Ser Val Pro Thr Ala

1 5 10 15

Gln Pro Gln Ala Glu Gly Ser Leu Ala Lys Ala Thr Thr Ala Pro Ala

20 25 30

Thr Thr Arg Asn Thr Gly Arg Gly Gly Glu Glu Lys Lys Lys Glu Lys

35 40 45

Glu Lys Glu Glu Gln Glu Glu Arg Glu Thr Lys Thr Pro Glu Cys Pro

50 55 60

Ser His Thr Gln Pro Leu Gly Val Tyr Leu Leu Thr Pro Ala Val Gln

65 70 75 80

Asp Leu Trp Leu Arg Asp Lys Ala Thr Phe Thr Cys Phe Val Val Gly

85 90 95

Ser Asp Leu Lys Asp Ala His Leu Thr Trp Glu Val Ala Gly Lys Val

100 105 110

Pro Thr Gly Gly Val Glu Glu Gly Leu Leu Glu Arg His Ser Asn Gly

115 120 125

Ser Gln Ser Gln His Ser Arg Leu Thr Leu Pro Arg Ser Leu Trp Asn

130 135 140

Ala Gly Thr Ser Val Thr Cys Thr Leu Asn His Pro Ser Leu Pro Pro

145 150 155 160

Gln Arg Leu Met Ala Leu Arg Glu Pro Ala Ala Gln Ala Pro Val Lys

165 170 175

Leu Ser Leu Asn Leu Leu Ala Ser Ser Asp Pro Pro Glu Ala Ala Ser

180 185 190

Trp Leu Leu Cys Glu Val Ser Gly Phe Ser Pro Pro Asn Ile Leu Leu

195 200 205

Met Trp Leu Glu Asp Gln Arg Glu Val Asn Thr Ser Gly Phe Ala Pro

210 215 220

Ala Arg Pro Pro Pro Gln Pro Gly Ser Thr Thr Phe Trp Ala Trp Ser

225 230 235 240

Val Leu Arg Val Pro Ala Pro Pro Ser Pro Gln Pro Ala Thr Tyr Thr

245 250 255

Cys Val Val Ser His Glu Asp Ser Arg Thr Leu Leu Asn Ala Ser Arg

260 265 270

Ser Leu Glu Val Ser Tyr Val Thr Asp His

275 280

<210> 6

<211> 24

<212> PRT

<213> Artificial sequence

<220>

<223> T2A

<400> 6

Leu Glu Gly Gly Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp

1 5 10 15

Val Glu Glu Asn Pro Gly Pro Arg

twenty

<210> 7

<211> 335

<212> PRT

<213> Artificial sequence

<220>

<223>tEGFR

<400> 7

Arg Lys Val Cys Asn Gly Ile Gly Ile Gly Glu Phe Lys Asp Ser Leu

1 5 10 15

Ser Ile Asn Ala Thr Asn Ile Lys His Phe Lys Asn Cys Thr Ser Ile

20 25 30

Ser Gly Asp Leu His Ile Leu Pro Val Ala Phe Arg Gly Asp Ser Phe

35 40 45

Thr His Thr Pro Pro Leu Asp Pro Gln Glu Leu Asp Ile Leu Lys Thr

50 55 60

Val Lys Glu Ile Thr Gly Phe Leu Leu Ile Gln Ala Trp Pro Glu Asn

65 70 75 80

Arg Thr Asp Leu His Ala Phe Glu Asn Leu Glu Ile Ile Arg Gly Arg

85 90 95

Thr Lys Gln His Gly Gln Phe Ser Leu Ala Val Val Ser Leu Asn Ile

100 105 110

Thr Ser Leu Gly Leu Arg Ser Leu Lys Glu Ile Ser Asp Gly Asp Val

115 120 125

Ile Ile Ser Gly Asn Lys Asn Leu Cys Tyr Ala Asn Thr Ile Asn Trp

130 135 140

Lys Lys Leu Phe Gly Thr Ser Gly Gln Lys Thr Lys Ile Ile Ser Asn

145 150 155 160

Arg Gly Glu Asn Ser Cys Lys Ala Thr Gly Gln Val Cys His Ala Leu

165 170 175

Cys Ser Pro Glu Gly Cys Trp Gly Pro Glu Pro Arg Asp Cys Val Ser

180 185 190

Cys Arg Asn Val Ser Arg Gly Arg Glu Cys Val Asp Lys Cys Asn Leu

195 200 205

Leu Glu Gly Glu Pro Arg Glu Phe Val Glu Asn Ser Glu Cys Ile Gln

210 215 220

Cys His Pro Glu Cys Leu Pro Gln Ala Met Asn Ile Thr Cys Thr Gly

225 230 235 240

Arg Gly Pro Asp Asn Cys Ile Gln Cys Ala His Tyr Ile Asp Gly Pro

245 250 255

His Cys Val Lys Thr Cys Pro Ala Gly Val Met Gly Glu Asn Asn Thr

260 265 270

Leu Val Trp Lys Tyr Ala Asp Ala Gly His Val Cys His Leu Cys His

275 280 285

Pro Asn Cys Thr Tyr Gly Cys Thr Gly Pro Gly Leu Glu Gly Cys Pro

290 295 300

Thr Asn Gly Pro Lys Ile Pro Ser Ile Ala Thr Gly Met Val Gly Ala

305 310 315 320

Leu Leu Leu Leu Leu Val Val Ala Leu Gly Ile Gly Leu Phe Met

325 330 335

<210> 8

<211> 27

<212> PRT

<213> Homo sapiens

<220>

<223>CD28

<300>

<308> UniProt P10747

<309> 1989-07-01

<400> 8

Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu

1 5 10 15

Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val

20 25

<210> 9

<211> 66

<212> PRT

<213> Homo sapiens

<220>

<223>CD28

<300>

<308> UniProt P10747

<309> 1989-07-01

<400> 9

Ile Glu Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser Asn

1 5 10 15

Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu

20 25 30

Phe Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly

35 40 45

Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe

50 55 60

Trp Val

65

<210> 10

<211> 41

<212> PRT

<213> Homo sapiens

<220>

<223>CD28

<300>

<308> UniProt P10747

<309> 1989-07-01

<400> 10

Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr

1 5 10 15

Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro

20 25 30

Pro Arg Asp Phe Ala Ala Tyr Arg Ser

35 40

<210> 11

<211> 41

<212> PRT

<213> Homo sapiens

<220>

<223>CD28

<400> 11

Arg Ser Lys Arg Ser Arg Gly Gly His Ser Asp Tyr Met Asn Met Thr

1 5 10 15

Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro

20 25 30

Pro Arg Asp Phe Ala Ala Tyr Arg Ser

35 40

<210> 12

<211> 42

<212> PRT

<213> Homo sapiens

<220>

<223> 4-1BB

<300>

<308> UniProt Q07011.1

<309> 1995-02-01

<400> 12

Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met

1 5 10 15

Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe

20 25 30

Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu

35 40

<210> 13

<211> 112

<212> PRT

<213> Homo sapiens

<220>

<223> CD3 zeta

<400> 13

Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly

1 5 10 15

Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr

20 25 30

Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys

35 40 45

Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys

50 55 60

Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg

65 70 75 80

Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala

85 90 95

Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

100 105 110

<210> 14

<211> 112

<212> PRT

<213> Homo sapiens

<220>

<223> CD3 zeta

<400> 14

Arg Val Lys Phe Ser Arg Ser Ala Glu Pro Pro Ala Tyr Gln Gln Gly

1 5 10 15

Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr

20 25 30

Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys

35 40 45

Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys

50 55 60

Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg

65 70 75 80

Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala

85 90 95

Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

100 105 110

<210> 15

<211> 112

<212> PRT

<213> Homo sapiens

<220>

<223> CD3 zeta

<400> 15

Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly

1 5 10 15

Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr

20 25 30

Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys

35 40 45

Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys

50 55 60

Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg

65 70 75 80

Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala

85 90 95

Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

100 105 110

<210> 16

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Linker

<220>

<221> REPEAT

<222> (5)...(9)

<223> SGGGG repeated 5 times

<400> 16

Pro Gly Gly Gly Ser Gly Gly Gly Gly

fifteen

<210> 17

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> Linker

<400> 17

Gly Ser Ala Asp Asp Ala Lys Lys Asp Ala Ala Lys Lys Asp Gly Lys

1 5 10 15

Ser

<210> 18

<211> 54

<212> PRT

<213> Homo sapiens

<220>

<223> Human BCMA intracellular domain

<400> 18

Met Leu Gln Met Ala Gly Gln Cys Ser Gln Asn Glu Tyr Phe Asp Ser

1 5 10 15

Leu Leu His Ala Cys Ile Pro Cys Gln Leu Arg Cys Ser Ser Asn Thr

20 25 30

Pro Pro Leu Thr Cys Gln Arg Tyr Cys Asn Ala Ser Val Thr Asn Ser

35 40 45

Val Lys Gly Thr Asn Ala

fifty

<210> 19

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> Linker

<400> 19

Gly Gly Gly Gly Ser

fifteen

<210> 20

<211> 231

<212> PRT

<213> Artificial sequence

<220>

<223> Modified human IgG1 Fc

<400> 20

Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro

1 5 10 15

Glu Ala Glu Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

20 25 30

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

35 40 45

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

50 55 60

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

65 70 75 80

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

85 90 95

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

100 105 110

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

115 120 125

Glu Pro Gln Val Tyr Thr Leu Pro Ser Arg Asp Glu Leu Thr Lys

130 135 140

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

145 150 155 160

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

165 170 175

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

180 185 190

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

195 200 205

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

210 215 220

Leu Ser Leu Ser Pro Gly Lys

225 230

<210> 21

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> CD33 signal peptide

<400> 21

Met Pro Leu Leu Leu Leu Leu Pro Leu Leu Trp Ala Gly Ala Leu Ala

1 5 10 15

<210> 22

<211> 306

<212> PRT

<213> Artificial sequence

<220>

<223> Fc BCMA

<400> 22

Met Pro Leu Leu Leu Leu Leu Pro Leu Leu Trp Ala Gly Ala Leu Ala

1 5 10 15

Met Leu Gln Met Ala Gly Gln Cys Ser Gln Asn Glu Tyr Phe Asp Ser

20 25 30

Leu Leu His Ala Cys Ile Pro Cys Gln Leu Arg Cys Ser Ser Asn Thr

35 40 45

Pro Pro Leu Thr Cys Gln Arg Tyr Cys Asn Ala Ser Val Thr Asn Ser

50 55 60

Val Lys Gly Thr Asn Ala Gly Gly Gly Gly Ser Pro Lys Ser Ser Asp

65 70 75 80

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Glu Gly Ala

85 90 95

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

100 105 110

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

115 120 125

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

130 135 140

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

145 150 155 160

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

165 170 175

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu

180 185 190

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

195 200 205

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

210 215 220

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

225 230 235 240

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

245 250 255

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

260 265 270

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

275 280 285

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

290 295 300

Gly Lys

305

<210> 23

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> T2A

<400> 23

Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro

1 5 10 15

GlyPro

<210> 24

<211> 22

<212> PRT

<213> Artificial sequence

<220>

<223> P2A

<400> 24

Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val

1 5 10 15

Glu Glu Asn Pro Gly Pro

twenty

<210> 25

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> P2A

<400> 25

Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn

1 5 10 15

Pro Gly Pro

<210> 26

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223>E2A

<400> 26

Gln Cys Thr Asn Tyr Ala Leu Leu Lys Leu Ala Gly Asp Val Glu Ser

1 5 10 15

Asn Pro Gly Pro

twenty

<210> 27

<211> 22

<212> PRT

<213> Artificial sequence

<220>

<223> F2A

<400> 27

Val Lys Gln Thr Leu Asn Phe Asp Leu Leu Lys Leu Ala Gly Asp Val

1 5 10 15

Glu Ser Asn Pro Gly Pro

twenty

<210> 28

<211> 335

<212> PRT

<213> Artificial sequence

<220>

<223>tEGFR

<400> 28

Arg Lys Val Cys Asn Gly Ile Gly Ile Gly Glu Phe Lys Asp Ser Leu

1 5 10 15

Ser Ile Asn Ala Thr Asn Ile Lys His Phe Lys Asn Cys Thr Ser Ile

20 25 30

Ser Gly Asp Leu His Ile Leu Pro Val Ala Phe Arg Gly Asp Ser Phe

35 40 45

Thr His Thr Pro Pro Leu Asp Pro Gln Glu Leu Asp Ile Leu Lys Thr

50 55 60

Val Lys Glu Ile Thr Gly Phe Leu Leu Ile Gln Ala Trp Pro Glu Asn

65 70 75 80

Arg Thr Asp Leu His Ala Phe Glu Asn Leu Glu Ile Ile Arg Gly Arg

85 90 95

Thr Lys Gln His Gly Gln Phe Ser Leu Ala Val Val Ser Leu Asn Ile

100 105 110

Thr Ser Leu Gly Leu Arg Ser Leu Lys Glu Ile Ser Asp Gly Asp Val

115 120 125

Ile Ile Ser Gly Asn Lys Asn Leu Cys Tyr Ala Asn Thr Ile Asn Trp

130 135 140

Lys Lys Leu Phe Gly Thr Ser Gly Gln Lys Thr Lys Ile Ile Ser Asn

145 150 155 160

Arg Gly Glu Asn Ser Cys Lys Ala Thr Gly Gln Val Cys His Ala Leu

165 170 175

Cys Ser Pro Glu Gly Cys Trp Gly Pro Glu Pro Arg Asp Cys Val Ser

180 185 190

Cys Arg Asn Val Ser Arg Gly Arg Glu Cys Val Asp Lys Cys Asn Leu

195 200 205

Leu Glu Gly Glu Pro Arg Glu Phe Val Glu Asn Ser Glu Cys Ile Gln

210 215 220

Cys His Pro Glu Cys Leu Pro Gln Ala Met Asn Ile Thr Cys Thr Gly

225 230 235 240

Arg Gly Pro Asp Asn Cys Ile Gln Cys Ala His Tyr Ile Asp Gly Pro

245 250 255

His Cys Val Lys Thr Cys Pro Ala Gly Val Met Gly Glu Asn Asn Thr

260 265 270

Leu Val Trp Lys Tyr Ala Asp Ala Gly His Val Cys His Leu Cys His

275 280 285

Pro Asn Cys Thr Tyr Gly Cys Thr Gly Pro Gly Leu Glu Gly Cys Pro

290 295 300

Thr Asn Gly Pro Lys Ile Pro Ser Ile Ala Thr Gly Met Val Gly Ala

305 310 315 320

Leu Leu Leu Leu Leu Val Val Ala Leu Gly Ile Gly Leu Phe Met

325 330 335

<210> 29

<211> 228

<212> PRT

<213> Homo sapiens

<220>

<223> Spacer Hinge-CH2-CH3

<400> 29

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Pro Val

1 5 10 15

Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

20 25 30

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

35 40 45

Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu

50 55 60

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Gln Ser Thr

65 70 75 80

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

85 90 95

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser

100 105 110

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

115 120 125

Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr

180 185 190

Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Leu Gly Lys

225

<210> 30

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 30

Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Ser Ile Asn Trp Val Lys Arg Ala Pro Gly Lys Gly Leu Lys Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp Phe

50 55 60

Arg Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Asn Asn Leu Lys Tyr Glu Asp Thr Ala Thr Tyr Phe Cys

85 90 95

Ala Leu Asp Tyr Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser

100 105 110

Val Thr Val Ser Ser

115

<210> 31

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 31

Asp Ile Val Leu Thr Gln Ser Pro Pro Ser Leu Ala Met Ser Leu Gly

1 5 10 15

Lys Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Thr Ile Leu

20 25 30

Gly Ser His Leu Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Thr Leu Leu Ile Gln Leu Ala Ser Asn Val Gln Thr Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asp

65 70 75 80

Pro Val Glu Glu Asp Asp Val Ala Val Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Thr Ile Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 32

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 32

Gln Ile Gln Leu Val Gln Ser Gly Pro Asp Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Phe

20 25 30

Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Phe Lys Trp Met

35 40 45

Ala Trp Ile Asn Thr Tyr Thr Gly Glu Ser Tyr Phe Ala Asp Asp Phe

50 55 60

Lys Gly Arg Phe Ala Phe Ser Val Glu Thr Ser Ala Thr Thr Ala Tyr

65 70 75 80

Leu Gln Ile Asn Asn Leu Lys Thr Glu Asp Thr Ala Thr Tyr Phe Cys

85 90 95

Ala Arg Gly Glu Ile Tyr Tyr Gly Tyr Asp Gly Gly Phe Ala Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ala

115 120

<210> 33

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 33

Asp Val Val Met Thr Gln Ser His Arg Phe Met Ser Thr Ser Val Gly

1 5 10 15

Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala

20 25 30

Val Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile

35 40 45

Phe Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly

50 55 60

Ser Gly Ser Gly Ala Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala

65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Asp Ile Lys

100 105

<210> 34

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 34

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Ile Ile Tyr Pro Gly Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe

50 55 60

Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Tyr Ser Gly Ser Phe Asp Asn Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 35

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 35

Ser Tyr Glu Leu Thr Gln Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Met Ser Cys Ser Gly Thr Ser Ser Asn Ile Gly Ser His

20 25 30

Ser Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Thr Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Gly Ser Leu

85 90 95

Asn Gly Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly

100 105 110

<210> 36

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 36

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Met Lys Lys Pro Gly Ala

1 5 10 15

Ser Leu Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ile Asp Tyr

20 25 30

Tyr Val Tyr Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Ser Met

35 40 45

Gly Trp Ile Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Ser Gln Arg Asp Gly Tyr Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 37

<211> 105

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 37

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Ala Ser Pro Gly Gln

1 5 10 15

Ser Ile Ala Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Trp Tyr

20 25 30

Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile Tyr Glu Asp Ser

35 40 45

Lys Arg Pro Ser Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly

50 55 60

Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala

65 70 75 80

Asp Tyr Tyr Cys Ser Ser Asn Thr Arg Ser Ser Thr Leu Val Phe Gly

85 90 95

Gly Gly Thr Lys Leu Thr Val Leu Gly

100 105

<210> 38

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 38

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Glu Asp Thr Ser Thr Asp Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ser Gly Tyr Ser Lys Ser Ile Val Ser Tyr Met Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 39

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 39

Leu Pro Val Leu Thr Gln Pro Ser Thr Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Val Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn

20 25 30

Val Val Phe Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Val

35 40 45

Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Val Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu

85 90 95

Ser Gly Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

<210> 40

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 40

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Ile Ile Pro Ile Leu Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ser Gly Tyr Gly Ser Tyr Arg Trp Glu Asp Ser Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 41

<211> 112

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 41

Gln Ala Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn

20 25 30

Tyr Val Phe Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu

85 90 95

Ser Ala Ser Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

<210> 42

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 42

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Glu Trp Met

35 40 45

Gly Trp Ile Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Asp Arg Ile Thr Val Thr Arg Asp Thr Ser Ser Asn Thr Gly Tyr

65 70 75 80

Met Glu Leu Thr Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ser Pro Tyr Ser Gly Val Leu Asp Lys Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 43

<211> 112

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 43

Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly

20 25 30

Phe Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Gly Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser

85 90 95

Leu Ser Gly Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

<210> 44

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> CDR H1 FMC63

<400> 44

Asp Tyr Gly Val Ser

fifteen

<210> 45

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> CDR H2 FMC63

<400> 45

Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser

1 5 10 15

<210> 46

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> CDR H3 FMC63

<400> 46

Tyr Ala Met Asp Tyr Trp Gly

fifteen

<210> 47

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> HC-CDR3 FMC63

<400> 47

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr

1 5 10

<210> 48

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> CDR L1 FMC63

<400> 48

Arg Ala Ser Gln Asp Ile Ser Lys Tyr Leu Asn

1 5 10

<210> 49

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> CDR L2 FMC63

<400> 49

Ser Arg Leu His Ser Gly Val

fifteen

<210> 50

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> LC-CDR2 FMC63

<400> 50

His Thr Ser Arg Leu His Ser

fifteen

<210> 51

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> CDR L3 FMC63

<400> 51

Gly Asn Thr Leu Pro Tyr Thr Phe Gly

fifteen

<210> 52

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> LC-CDR3 FMC63

<400> 52

Gln Gln Gly Asn Thr Leu Pro Tyr Thr

fifteen

<210> 53

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> VH FMC63

<400> 53

Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln

1 5 10 15

Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr

20 25 30

Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu

65 70 75 80

Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala

85 90 95

Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 54

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> VL FMC63

<400> 54

Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln

65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr

100 105

<210> 55

<211> 245

<212> PRT

<213> Artificial sequence

<220>

<223> scFv FMC63

<400> 55

Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile

35 40 45

Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln

65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr Gly Ser Thr Ser Gly

100 105 110

Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Glu Val Lys

115 120 125

Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln Ser Leu Ser

130 135 140

Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val Ser

145 150 155 160

Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu Gly Val Ile

165 170 175

Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser Arg Leu

180 185 190

Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu Lys Met Asn

195 200 205

Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala Lys His Tyr

210 215 220

Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser

225 230 235 240

Val Thr Val Ser Ser

245

<210> 56

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> CDR L1 SJ25C1

<400> 56

Lys Ala Ser Gln Asn Val Gly Thr Asn Val Ala

1 5 10

<210> 57

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> CDR L2 SJ25C1

<400> 57

Ser Ala Thr Tyr Arg Asn Ser

fifteen

<210> 58

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> CDR L3 SJ25C1

<400> 58

Gln Gln Tyr Asn Arg Tyr Pro Tyr Thr

fifteen

<210> 59

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> CDR H1 SJ25C1

<400> 59

Ser Tyr Trp Met Asn

fifteen

<210> 60

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> CDR H2 SJ25C1

<400> 60

Gln Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys

1 5 10 15

gly

<210> 61

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> CDR H3 SJ25C1

<400> 61

Lys Thr Ile Ser Ser Val Val Asp Phe Tyr Phe Asp Tyr

1 5 10

<210> 62

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> VH SJ25C1

<400> 62

Glu Val Lys Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser

1 5 10 15

Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr

20 25 30

Trp Met Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe

50 55 60

Lys Gly Gln Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys

85 90 95

Ala Arg Lys Thr Ile Ser Ser Val Val Asp Phe Tyr Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 63

<211> 108

<212> PRT

<213> Artificial sequence

<220>

<223> VL SJ25C1

<400> 63

Asp Ile Glu Leu Thr Gln Ser Pro Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Pro Leu Ile

35 40 45

Tyr Ser Ala Thr Tyr Arg Asn Ser Gly Val Pro Asp Arg Phe Thr Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Asn Val Gln Ser

65 70 75 80

Lys Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr Asn Arg Tyr Pro Tyr

85 90 95

Thr Ser Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg

100 105

<210> 64

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> Linker

<400> 64

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10 15

<210> 65

<211> 245

<212> PRT

<213> Artificial sequence

<220>

<223> scFv SJ25C1

<400> 65

Glu Val Lys Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser

1 5 10 15

Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr

20 25 30

Trp Met Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe

50 55 60

Lys Gly Gln Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys

85 90 95

Ala Arg Lys Thr Ile Ser Ser Val Val Asp Phe Tyr Phe Asp Tyr Trp

100 105 110

Gly Glyn Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser

130 135 140

Pro Lys Phe Met Ser Thr Ser Val Gly Asp Arg Val Ser Val Thr Cys

145 150 155 160

Lys Ala Ser Gln Asn Val Gly Thr Asn Val Ala Trp Tyr Gln Gln Lys

165 170 175

Pro Gly Gln Ser Pro Lys Pro Leu Ile Tyr Ser Ala Thr Tyr Arg Asn

180 185 190

Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe

195 200 205

Thr Leu Thr Ile Thr Asn Val Gln Ser Lys Asp Leu Ala Asp Tyr Phe

210 215 220

Cys Gln Gln Tyr Asn Arg Tyr Pro Tyr Thr Ser Gly Gly Gly Thr Lys

225 230 235 240

Leu Glu Ile Lys Arg

245

<210> 66

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> HC-CDR3 FMC63

<400> 66

His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr

1 5 10

<210> 67

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> LC-CDR2 FMC63

<400> 67

His Thr Ser Arg Leu His Ser

fifteen

<210> 68

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> LC-CDR3 FMC63

<400> 68

Gln Gln Gly Asn Thr Leu Pro Tyr Thr

fifteen

<210> 69

<211> 735

<212> DNA

<213> Artificial sequence

<220>

<223>scFv

<400> 69

gacatccaga tgacccagac cacctccagc ctgagcgcca gcctgggcga ccgggtgacc 60

atcagctgcc gggccagcca ggacatcagc aagtacctga actggtatca gcagaagccc 120

gacggcaccg tcaagctgct gatctaccac accagccggc tgcacagcgg cgtgcccagc 180

cggtttagcg gcagcggctc cggcaccgac tacagcctga ccatctccaa cctggaacag 240

gaagatatcg ccacctactt ttgccagcag ggcaacacac tgccctacac ctttggcggc 300

ggaacaaagc tggaaatcac cggcagcacc tccggcagcg gcaagcctgg cagcggcgag 360

ggcagcacca agggcgaggt gaagctgcag gaaagcggcc ctggcctggt ggcccccagc 420

cagagcctga gcgtgacctg caccgtgagc ggcgtgagcc tgcccgacta cggcgtgagc 480

tggatccggc agccccccag gaagggcctg gaatggctgg gcgtgatctg gggcagcgag 540

accacctact acaacagcgc cctgaagagc cggctgacca tcatcaagga caacagcaag 600

agccaggtgt tcctgaagat gaacagcctg cagaccgacg acaccgccat ctactactgc 660

gccaagcact actactacgg cggcagctac gccatggact actggggcca gggcaccagc 720

gtgaccgtga gcagc 735

<210> 70

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> Linker

<400> 70

Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr

1 5 10 15

Lys Gly

<210> 71

<211> 4

<212> PRT

<213> Artificial sequence

<220>

<223> Linker

<400> 71

Gly Gly Gly Ser

one

<210> 72

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> Linker

<400> 72

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10 15

<210> 73

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> Linker

<400> 73

Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr

1 5 10 15

Lys Gly

<210> 74

<211> 21

<212> PRT

<213> Artificial sequence

<220>

<223> Linker

<400> 74

Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

1 5 10 15

Ser Leu Glu Met Ala

twenty

<210> 75

<211> 21

<212> PRT

<213> Artificial sequence

<220>

<223> CD8a signal peptide

<400> 75

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro

twenty

<210> 76

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> Signal peptide

<400> 76

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly

twenty

<210> 77

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 77

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ala Glu Met Gly Ala Val Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Met Val Thr Val Ser Ser

115

<210> 78

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 78

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ile Ser Trp Pro Phe

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 79

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 79

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Gly Thr Tyr Leu Gly Gly Leu Trp Tyr Phe Asp Leu Trp

100 105 110

Gly Arg Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 80

<211> 112

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 80

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gly

85 90 95

Leu Gly Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 81

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 81

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Glu Trp Met

35 40 45

Gly Ile Ile Asn Pro Gly Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Ser Trp Pro Met Asp Val Trp Gly Gln Gly Thr Thr Val

100 105 110

Thr Val Ser Ser

115

<210> 82

<211> 106

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 82

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ala Ala Tyr Pro Thr

85 90 95

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 83

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 83

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Gly Arg Gly Tyr Ala Thr Ser Leu Ala Phe Asp Ile Trp

100 105 110

Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 84

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 84

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg His Val Trp Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 85

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 85

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ser Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Thr Ile Ser Ser Ser Ser Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Ser Gln Glu His Leu Ile Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 86

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 86

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Phe Tyr Tyr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 87

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 87

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Asp Phe Trp Ser Gly Ser Pro Pro Gly Leu Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 88

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 88

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ile Tyr Thr Phe Pro Phe

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 89

<211> 126

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 89

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Pro Glu Tyr Ser Ser Ser Ile Trp His Tyr Tyr Tyr Gly

100 105 110

Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120 125

<210> 90

<211> 113

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 90

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

20 25 30

Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Phe Ala His Thr Pro Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 91

<211> 123

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 91

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Lys Gly Pro Leu Gln Glu Pro Pro Tyr Asp Tyr Gly Met Asp Val

100 105 110

Trp Gly Glyn Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 92

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 92

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Ser Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln His His Val Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 93

<211> 121

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 93

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Tyr Tyr Ser His Asp Met Trp Ser Glu Asp Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 94

<211> 112

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 94

Leu Pro Val Leu Thr Gln Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ser Gly Arg Ser Ser Asn Ile Gly Ser Asn

20 25 30

Ser Val Asn Trp Tyr Arg Gln Leu Pro Gly Ala Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Ser Asn Asn Gln Arg Pro Pro Gly Val Pro Val Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Thr Tyr Tyr Cys Ala Thr Trp Asp Asp Asn Leu

85 90 95

Asn Val His Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

<210> 95

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 95

Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Ser Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp Phe

50 55 60

Arg Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Tyr Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 96

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 96

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Ser Val Ile

20 25 30

Gly Ala His Leu Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Thr Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Ala Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Ile Phe Pro Arg Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 97

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 97

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ala Leu Ser Asn His

20 25 30

Gly Met Ser Trp Val Arg Arg Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Val Tyr Ser Gly Ser Thr Tyr Tyr Ala Ala Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ser

85 90 95

Ala His Gly Gly Glu Ser Asp Val Trp Gly Gln Gly Thr Thr Val Thr

100 105 110

Val Ser Ser

115

<210> 98

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 98

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 99

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 99

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Gly Trp Val

35 40 45

Ser Gly Ile Ser Arg Ser Gly Glu Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ser Pro Ala His Tyr Tyr Gly Gly Met Asp Val Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 100

<211> 109

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 100

Asp Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Ser

20 25 30

Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Arg Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Ser Ala Val Tyr Tyr Cys Gln Gln Tyr His Ser Ser Pro

85 90 95

Ser Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 101

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 101

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ala Leu Ser Asn His

20 25 30

Gly Met Ser Trp Val Arg Arg Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Val Tyr Ser Gly Ser Thr Tyr Tyr Ala Ala Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ser

85 90 95

Ala His Gly Gly Glu Ser Asp Val Trp Gly Gln Gly Thr Thr Val Thr

100 105 110

Val Ser Ser

115

<210> 102

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 102

Asp Ile Arg Leu Thr Gln Ser Pro Ser Pro Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Glu Asp Ile Asn Lys Phe

20 25 30

Leu Asn Trp Tyr His Gln Thr Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Thr Leu Gln Thr Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Gly Thr Tyr Tyr Cys Gln Gln Tyr Glu Ser Leu Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 103

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 103

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ala Leu Ser Asn His

20 25 30

Gly Met Ser Trp Val Arg Arg Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Val Tyr Ser Gly Ser Thr Tyr Tyr Ala Ala Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ser

85 90 95

Ala His Gly Gly Glu Ser Asp Val Trp Gly Gln Gly Thr Thr Val Thr

100 105 110

Val Ser Ser

115

<210> 104

<211> 109

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 104

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Gly Ser Ser

20 25 30

Ser Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Met Tyr Gly Ala Ser Ser Arg Ala Ser Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ala Gly Ser Pro

85 90 95

Pro Phe Thr Phe Gly Glyn Gly Thr Lys Val Glu Ile Lys

100 105

<210> 105

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 105

Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Arg His Tyr

20 25 30

Ser Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met

35 40 45

Gly Arg Ile Asn Thr Glu Ser Gly Val Pro Ile Tyr Ala Asp Asp Phe

50 55 60

Lys Gly Arg Phe Ala Phe Ser Val Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Val Ile Asn Asn Leu Lys Asp Glu Asp Thr Ala Ser Tyr Phe Cys

85 90 95

Ser Asn Asp Tyr Leu Tyr Ser Leu Asp Phe Trp Gly Gln Gly Thr Ala

100 105 110

Leu Thr Val Ser Ser

115

<210> 106

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 106

Asp Ile Val Leu Thr Gln Ser Pro Pro Ser Leu Ala Met Ser Leu Gly

1 5 10 15

Lys Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Thr Ile Leu

20 25 30

Gly Ser His Leu Ile Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Thr Leu Leu Ile Gln Leu Ala Ser Asn Val Gln Thr Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asp

65 70 75 80

Pro Val Glu Glu Asp Asp Val Ala Val Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Thr Ile Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 107

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 107

Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr His Tyr

20 25 30

Ser Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met

35 40 45

Gly Arg Ile Asn Thr Glu Thr Gly Glu Pro Leu Tyr Ala Asp Asp Phe

50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Val Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Phe Phe Cys

85 90 95

Ser Asn Asp Tyr Leu Tyr Ser Cys Asp Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Leu Thr Val Ser Ser

115

<210> 108

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 108

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Met Ser Leu Gly

1 5 10 15

Lys Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Ser Val Ile

20 25 30

Gly Ala His Leu Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Thr Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asp

65 70 75 80

Pro Val Glu Glu Asp Asp Val Ala Ile Tyr Ser Cys Leu Gln Ser Arg

85 90 95

Ile Phe Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 109

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 109

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Pro Asp Tyr

20 25 30

Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Tyr Phe Ala Ser Gly Asn Ser Glu Tyr Asn Gln Lys Phe

50 55 60

Thr Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Asn Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Ser Leu Tyr Asp Tyr Asp Trp Tyr Phe Asp Val Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 110

<211> 112

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 110

Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Ile Tyr Tyr Cys Ser Gln Ser

85 90 95

Ser Ile Tyr Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 111

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 111

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Pro Asp Tyr

20 25 30

Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Tyr Phe Ala Ser Gly Asn Ser Glu Tyr Asn Gln Lys Phe

50 55 60

Thr Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Ser Leu Tyr Asp Tyr Asp Trp Tyr Phe Asp Val Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 112

<211> 112

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 112

Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Ala Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ala Glu Thr

85 90 95

Ser His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 113

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> sdAb anti-BCMA

<400> 113

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Leu Asp Tyr Tyr

20 25 30

Ala Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val

35 40 45

Ile Cys Ile Ser Arg Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Thr Val Tyr

65 70 75 80

Leu Gln Met Ile Ser Leu Lys Pro Glu Asp Thr Ala Ala Tyr Tyr Cys

85 90 95

Ala Ala Gly Ala Asp Cys Ser Gly Tyr Leu Arg Asp Tyr Glu Phe Arg

100 105 110

Gly Gln Gly Thr Gln Val Thr Val Ser Ser

115 120

<210> 114

<211> 39

<212> PRT

<213> Artificial sequence

<220>

<223> Spacer CD28

<400> 114

Ile Glu Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser Asn

1 5 10 15

Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu

20 25 30

Phe Pro Gly Pro Ser Lys Pro

35

<210> 115

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223>TM CD8a

<400> 115

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu

1 5 10 15

Ser Leu Val Ile Thr Leu Tyr Cys Asn

20 25

<210> 116

<211> 30

<212> PRT

<213> Artificial sequence

<220>

<223> Spacer CD28 (truncated)

<400> 116

Leu Asp Asn Glu Lys Ser Asn Gly Thr Ile Ile His Val Lys Gly Lys

1 5 10 15

His Leu Cys Pro Ser Pro Leu Phe Pro Gly Pro Ser Lys Pro

20 25 30

<210> 117

<211> 46

<212> PRT

<213> Artificial sequence

<220>

<223> Hinge CD8a

<400> 117

Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile

1 5 10 15

Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala

20 25 30

Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp

35 40 45

<210> 118

<211> 45

<212> PRT

<213> Artificial sequence

<220>

<223> Hinge CD8a

<400> 118

Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala

1 5 10 15

Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly

20 25 30

Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp

35 40 45

<210> 119

<211> 55

<212> PRT

<213> Artificial sequence

<220>

<223> Hinge CD8a

<400> 119

Phe Val Pro Val Phe Leu Pro Ala Lys Pro Thr Thr Thr Pro Ala Pro

1 5 10 15

Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu

20 25 30

Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg

35 40 45

Gly Leu Asp Phe Ala Cys Asp

50 55

<210> 120

<211> 39

<212> PRT

<213> Artificial sequence

<220>

<223> Hinge CTLA4

<400> 120

Asp Thr Gly Leu Tyr Ile Cys Lys Val Glu Leu Met Tyr Pro Pro Pro

1 5 10 15

Tyr Tyr Leu Gly Ile Gly Asn Gly Thr Gln Ile Tyr Val Ile Asp Pro

20 25 30

Glu Pro Cys Pro Asp Ser Asp

35

<210> 121

<211> 24

<212> PRT

<213> Artificial sequence

<220>

<223> CTLA4TM

<400> 121

Phe Leu Leu Trp Ile Leu Ala Ala Val Ser Ser Gly Leu Phe Phe Tyr

1 5 10 15

Ser Phe Leu Leu Thr Ala Val Ser

twenty

<210> 122

<211> 38

<212> PRT

<213> Artificial sequence

<220>

<223> Hinge PD-1

<400> 122

Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg

1 5 10 15

Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro Arg Pro Ala Gly

20 25 30

Gln Phe Gln Thr Leu Val

35

<210> 123

<211> 21

<212> PRT

<213> Artificial sequence

<220>

<223>PD-1TM

<400> 123

Val Gly Val Val Gly Gly Leu Leu Gly Ser Leu Val Leu Leu Val Trp

1 5 10 15

Val Leu Ala Val Ile

twenty

<210> 124

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> Hinge Fc(gamma) RIIIa

<400> 124

Gly Leu Ala Val Ser Thr Ile Ser Ser Phe Phe Pro Pro Gly Tyr Gln

1 5 10 15

<210> 125

<211> 231

<212> PRT

<213> Artificial sequence

<220>

<223> IgG1 hinge

<400> 125

Glu Pro Lys Ser Pro Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala

1 5 10 15

Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

20 25 30

Asp Thr Leu Met Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val

35 40 45

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

50 55 60

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

65 70 75 80

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

85 90 95

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

100 105 110

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

115 120 125

Glu Pro Gln Val Tyr Thr Leu Pro Ser Arg Asp Glu Leu Thr Lys

130 135 140

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

145 150 155 160

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

165 170 175

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

180 185 190

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

195 200 205

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

210 215 220

Leu Ser Leu Ser Pro Gly Lys

225 230

<210> 126

<211> 457

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 126

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ala Glu Met Gly Ala Val Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Met Val Thr Val Ser Ser Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly

115 120 125

Ser Gly Glu Gly Ser Thr Lys Gly Glu Ile Val Leu Thr Gln Ser Pro

130 135 140

Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg

145 150 155 160

Ala Ser Gln Ser Val Ser Arg Tyr Leu Ala Trp Tyr Gln Gln Lys Pro

165 170 175

Gly Gln Ala Pro Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr

180 185 190

Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

195 200 205

Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys

210 215 220

Gln Gln Arg Ile Ser Trp Pro Phe Thr Phe Gly Gly Gly Thr Lys Val

225 230 235 240

Glu Ile Lys Arg Ala Ala Ala Leu Asp Asn Glu Lys Ser Asn Gly Thr

245 250 255

Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe Pro

260 265 270

Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val Leu

275 280 285

Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val

290 295 300

Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr

305 310 315 320

Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro

325 330 335

Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser

340 345 350

Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu

355 360 365

Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg

370 375 380

Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln

385 390 395 400

Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr

405 410 415

Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp

420 425 430

Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala

435 440 445

Leu His Met Gln Ala Leu Pro Pro Arg

450 455

<210> 127

<211> 457

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 127

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ile Ser Trp Pro Phe

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Gly Ser Thr Ser

100 105 110

Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Glu Val

115 120 125

Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu

130 135 140

Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ala Met

145 150 155 160

Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ala

165 170 175

Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly

180 185 190

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

195 200 205

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg

210 215 220

Ala Glu Met Gly Ala Val Phe Asp Ile Trp Gly Gln Gly Thr Met Val

225 230 235 240

Thr Val Ser Ser Ala Ala Ala Leu Asp Asn Glu Lys Ser Asn Gly Thr

245 250 255

Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe Pro

260 265 270

Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val Leu

275 280 285

Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val

290 295 300

Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr

305 310 315 320

Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro

325 330 335

Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser

340 345 350

Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu

355 360 365

Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg

370 375 380

Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln

385 390 395 400

Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr

405 410 415

Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp

420 425 430

Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala

435 440 445

Leu His Met Gln Ala Leu Pro Pro Arg

450 455

<210> 128

<211> 466

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 128

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Gly Thr Tyr Leu Gly Gly Leu Trp Tyr Phe Asp Leu Trp

100 105 110

Gly Arg Gly Thr Leu Val Thr Val Ser Ser Gly Ser Thr Ser Gly Ser

115 120 125

Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Asp Ile Val Met

130 135 140

Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly Glu Pro Ala Ser

145 150 155 160

Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser Asn Gly Tyr Asn

165 170 175

Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu

180 185 190

Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro Asp Arg Phe Ser

195 200 205

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu

210 215 220

Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gly Leu Gly Leu Pro

225 230 235 240

Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Ala Ala Ala

245 250 255

Leu Asp Asn Glu Lys Ser Asn Gly Thr Ile Ile His Val Lys Gly Lys

260 265 270

His Leu Cys Pro Ser Pro Leu Phe Pro Gly Pro Ser Lys Pro Phe Trp

275 280 285

Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val

290 295 300

Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Leu

305 310 315 320

Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr

325 330 335

Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr

340 345 350

Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln

355 360 365

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

370 375 380

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

385 390 395 400

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

405 410 415

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

420 425 430

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

435 440 445

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

450 455 460

Pro Arg

465

<210> 129

<211> 466

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 129

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gly

85 90 95

Leu Gly Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

Arg Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser

115 120 125

Thr Lys Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln

130 135 140

Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe

145 150 155 160

Ser Ser Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu

165 170 175

Glu Trp Val Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala

180 185 190

Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn

195 200 205

Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val

210 215 220

Tyr Tyr Cys Ala Arg Asp Gly Thr Tyr Leu Gly Gly Leu Trp Tyr Phe

225 230 235 240

Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser Ala Ala Ala

245 250 255

Leu Asp Asn Glu Lys Ser Asn Gly Thr Ile Ile His Val Lys Gly Lys

260 265 270

His Leu Cys Pro Ser Pro Leu Phe Pro Gly Pro Ser Lys Pro Phe Trp

275 280 285

Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val

290 295 300

Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Leu

305 310 315 320

Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr

325 330 335

Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr

340 345 350

Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln

355 360 365

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

370 375 380

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

385 390 395 400

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

405 410 415

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

420 425 430

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

435 440 445

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

450 455 460

Pro Arg

465

<210> 130

<211> 454

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 130

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Glu Trp Met

35 40 45

Gly Ile Ile Asn Pro Gly Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Ser Trp Pro Met Asp Val Trp Gly Gln Gly Thr Thr Val

100 105 110

Thr Val Ser Ser Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly

115 120 125

Glu Gly Ser Thr Lys Gly Glu Ile Val Met Thr Gln Ser Pro Ala Thr

130 135 140

Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser

145 150 155 160

Gln Ser Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

165 170 175

Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile

180 185 190

Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr

195 200 205

Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln

210 215 220

Tyr Ala Ala Tyr Pro Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

225 230 235 240

Arg Ala Ala Ala Leu Asp Asn Glu Lys Ser Asn Gly Thr Ile Ile His

245 250 255

Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe Pro Gly Pro Ser

260 265 270

Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr

275 280 285

Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys

290 295 300

Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg

305 310 315 320

Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp

325 330 335

Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala

340 345 350

Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu

355 360 365

Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp

370 375 380

Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu

385 390 395 400

Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile

405 410 415

Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr

420 425 430

Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met

435 440 445

Gln Ala Leu Pro Pro Arg

450

<210> 131

<211> 454

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 131

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ala Ala Tyr Pro Thr

85 90 95

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Gly Ser Thr Ser Gly

100 105 110

Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val Gln

115 120 125

Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys

130 135 140

Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Tyr Met His

145 150 155 160

Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly Ile Ile

165 170 175

Asn Pro Gly Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe Gln Gly Arg

180 185 190

Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr Met Glu Leu

195 200 205

Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu

210 215 220

Ser Trp Pro Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser

225 230 235 240

Ser Ala Ala Ala Leu Asp Asn Glu Lys Ser Asn Gly Thr Ile Ile His

245 250 255

Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe Pro Gly Pro Ser

260 265 270

Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr

275 280 285

Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys

290 295 300

Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg

305 310 315 320

Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp

325 330 335

Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala

340 345 350

Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu

355 360 365

Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp

370 375 380

Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu

385 390 395 400

Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile

405 410 415

Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr

420 425 430

Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met

435 440 445

Gln Ala Leu Pro Pro Arg

450

<210> 132

<211> 461

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 132

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Gly Arg Gly Tyr Ala Thr Ser Leu Ala Phe Asp Ile Trp

100 105 110

Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Ser Thr Ser Gly Ser

115 120 125

Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Glu Ile Val Leu

130 135 140

Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr

145 150 155 160

Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala Trp Tyr

165 170 175

Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Asp Ala Ser

180 185 190

Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly

195 200 205

Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala

210 215 220

Val Tyr Tyr Cys Gln Gln Arg His Val Trp Pro Pro Thr Phe Gly Gly

225 230 235 240

Gly Thr Lys Val Glu Ile Lys Arg Ala Ala Ala Leu Asp Asn Glu Lys

245 250 255

Ser Asn Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser

260 265 270

Pro Leu Phe Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val

275 280 285

Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile

290 295 300

Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr

305 310 315 320

Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln

325 330 335

Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys

340 345 350

Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln

355 360 365

Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu

370 375 380

Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg

385 390 395 400

Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met

405 410 415

Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly

420 425 430

Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp

435 440 445

Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455 460

<210> 133

<211> 461

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 133

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg His Val Trp Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Gly Ser Thr Ser

100 105 110

Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Leu

115 120 125

Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu

130 135 140

Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser Ser Tyr

145 150 155 160

Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

165 170 175

Gly Ser Ile Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys

180 185 190

Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

195 200 205

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

210 215 220

Arg Gly Arg Gly Tyr Ala Thr Ser Leu Ala Phe Asp Ile Trp Gly Gln

225 230 235 240

Gly Thr Met Val Thr Val Ser Ser Ala Ala Ala Leu Asp Asn Glu Lys

245 250 255

Ser Asn Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser

260 265 270

Pro Leu Phe Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val

275 280 285

Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile

290 295 300

Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr

305 310 315 320

Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln

325 330 335

Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys

340 345 350

Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln

355 360 365

Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu

370 375 380

Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg

385 390 395 400

Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met

405 410 415

Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly

420 425 430

Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp

435 440 445

Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455 460

<210> 134

<211> 458

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 134

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ser Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Thr Ile Ser Ser Ser Ser Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Ser Gln Glu His Leu Ile Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Gly Ser Thr Ser Gly Ser Gly Lys Pro

115 120 125

Gly Ser Gly Glu Gly Ser Thr Lys Gly Glu Ile Val Leu Thr Gln Ser

130 135 140

Pro Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys

145 150 155 160

Arg Ala Ser Gln Ser Val Ser Arg Tyr Leu Ala Trp Tyr Gln Gln Lys

165 170 175

Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala

180 185 190

Thr Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe

195 200 205

Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr

210 215 220

Cys Gln Gln Arg Phe Tyr Tyr Pro Trp Thr Phe Gly Gly Gly Thr Lys

225 230 235 240

Val Glu Ile Lys Arg Ala Ala Ala Leu Asp Asn Glu Lys Ser Asn Gly

245 250 255

Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe

260 265 270

Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val

275 280 285

Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp

290 295 300

Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met

305 310 315 320

Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala

325 330 335

Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg

340 345 350

Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn

355 360 365

Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg

370 375 380

Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro

385 390 395 400

Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala

405 410 415

Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His

420 425 430

Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp

435 440 445

Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455

<210> 135

<211> 458

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 135

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Phe Tyr Tyr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Gly Ser Thr Ser

100 105 110

Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Glu Val

115 120 125

Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu

130 135 140

Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ser Met

145 150 155 160

Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Thr

165 170 175

Ile Ser Ser Ser Ser Ser Thr Ile Tyr Tyr Ala Asp Ser Val Lys Gly

180 185 190

Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln

195 200 205

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg

210 215 220

Gly Ser Gln Glu His Leu Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu

225 230 235 240

Val Thr Val Ser Ser Ala Ala Ala Leu Asp Asn Glu Lys Ser Asn Gly

245 250 255

Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe

260 265 270

Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val

275 280 285

Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp

290 295 300

Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met

305 310 315 320

Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala

325 330 335

Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg

340 345 350

Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn

355 360 365

Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg

370 375 380

Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro

385 390 395 400

Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala

405 410 415

Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His

420 425 430

Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp

435 440 445

Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455

<210> 136

<211> 461

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 136

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Asp Phe Trp Ser Gly Ser Pro Pro Gly Leu Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Thr Ser Gly Ser

115 120 125

Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Asp Ile Gln Leu

130 135 140

Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly Asp Arg Val Thr

145 150 155 160

Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Ala Trp Tyr

165 170 175

Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Gly Ala Ser

180 185 190

Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly

195 200 205

Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala

210 215 220

Thr Tyr Tyr Cys Gln Gln Ile Tyr Thr Phe Pro Phe Thr Phe Gly Gly

225 230 235 240

Gly Thr Lys Val Glu Ile Lys Arg Ala Ala Ala Leu Asp Asn Glu Lys

245 250 255

Ser Asn Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser

260 265 270

Pro Leu Phe Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val

275 280 285

Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile

290 295 300

Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr

305 310 315 320

Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln

325 330 335

Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys

340 345 350

Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln

355 360 365

Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu

370 375 380

Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg

385 390 395 400

Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met

405 410 415

Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly

420 425 430

Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp

435 440 445

Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455 460

<210> 137

<211> 461

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 137

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ile Tyr Thr Phe Pro Phe

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Gly Ser Thr Ser

100 105 110

Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val

115 120 125

Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu

130 135 140

Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Gly Met

145 150 155 160

His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Val

165 170 175

Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys Gly

180 185 190

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

195 200 205

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg

210 215 220

Thr Asp Phe Trp Ser Gly Ser Pro Pro Gly Leu Asp Tyr Trp Gly Gln

225 230 235 240

Gly Thr Leu Val Thr Val Ser Ser Ala Ala Ala Leu Asp Asn Glu Lys

245 250 255

Ser Asn Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser

260 265 270

Pro Leu Phe Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val

275 280 285

Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile

290 295 300

Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr

305 310 315 320

Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln

325 330 335

Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys

340 345 350

Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln

355 360 365

Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu

370 375 380

Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg

385 390 395 400

Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met

405 410 415

Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly

420 425 430

Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp

435 440 445

Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455 460

<210> 138

<211> 471

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 138

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Pro Glu Tyr Ser Ser Ser Ile Trp His Tyr Tyr Tyr Gly

100 105 110

Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Ser

115 120 125

Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly

130 135 140

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

145 150 155 160

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

165 170 175

Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

180 185 190

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

195 200 205

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

210 215 220

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

225 230 235 240

Phe Ala His Thr Pro Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile

245 250 255

Lys Arg Ala Ala Ala Leu Asp Asn Glu Lys Ser Asn Gly Thr Ile Ile

260 265 270

His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe Pro Gly Pro

275 280 285

Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys

290 295 300

Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser

305 310 315 320

Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg

325 330 335

Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg

340 345 350

Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp

355 360 365

Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn

370 375 380

Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg

385 390 395 400

Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly

405 410 415

Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu

420 425 430

Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu

435 440 445

Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His

450 455 460

Met Gln Ala Leu Pro Pro Arg

465 470

<210> 139

<211> 471

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 139

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

20 25 30

Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Phe Ala His Thr Pro Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly

115 120 125

Ser Thr Lys Gly Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys

130 135 140

Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr

145 150 155 160

Phe Ser Ser Tyr Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly

165 170 175

Leu Glu Trp Met Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr

180 185 190

Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr

195 200 205

Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala

210 215 220

Val Tyr Tyr Cys Ala Arg Thr Pro Glu Tyr Ser Ser Ser Ile Trp His

225 230 235 240

Tyr Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val

245 250 255

Ser Ser Ala Ala Ala Leu Asp Glu Lys Ser Asn Gly Thr Ile Ile

260 265 270

His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe Pro Gly Pro

275 280 285

Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys

290 295 300

Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser

305 310 315 320

Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg

325 330 335

Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg

340 345 350

Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp

355 360 365

Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn

370 375 380

Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg

385 390 395 400

Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly

405 410 415

Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu

420 425 430

Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu

435 440 445

Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His

450 455 460

Met Gln Ala Leu Pro Pro Arg

465 470

<210> 140

<211> 462

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 140

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Lys Gly Pro Leu Gln Glu Pro Pro Tyr Asp Tyr Gly Met Asp Val

100 105 110

Trp Gly Glyn Gly Thr Thr Val Thr Val Ser Ser Gly Ser Thr Ser Gly

115 120 125

Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Glu Ile Val

130 135 140

Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala

145 150 155 160

Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala Trp

165 170 175

Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Ser Ala

180 185 190

Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser

195 200 205

Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe

210 215 220

Ala Val Tyr Tyr Cys Gln Gln His His Val Trp Pro Leu Thr Phe Gly

225 230 235 240

Gly Gly Thr Lys Val Glu Ile Lys Arg Ala Ala Ala Leu Asp Asn Glu

245 250 255

Lys Ser Asn Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro

260 265 270

Ser Pro Leu Phe Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val

275 280 285

Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe

290 295 300

Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp

305 310 315 320

Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr

325 330 335

Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val

340 345 350

Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn

355 360 365

Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val

370 375 380

Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg

385 390 395 400

Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys

405 410 415

Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg

420 425 430

Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys

435 440 445

Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455 460

<210> 141

<211> 462

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 141

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Ser Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln His His Val Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Gly Ser Thr Ser

100 105 110

Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val

115 120 125

Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu

130 135 140

Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Gly Met

145 150 155 160

His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Val

165 170 175

Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys Gly

180 185 190

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

195 200 205

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Val Lys

210 215 220

Gly Pro Leu Gln Glu Pro Pro Tyr Asp Tyr Gly Met Asp Val Trp Gly

225 230 235 240

Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ala Ala Leu Asp Asn Glu

245 250 255

Lys Ser Asn Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro

260 265 270

Ser Pro Leu Phe Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val

275 280 285

Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe

290 295 300

Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp

305 310 315 320

Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr

325 330 335

Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val

340 345 350

Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn

355 360 365

Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val

370 375 380

Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg

385 390 395 400

Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys

405 410 415

Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg

420 425 430

Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys

435 440 445

Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455 460

<210> 142

<211> 466

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 142

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Ala Ser Pro Gly Gln

1 5 10 15

Ser Ile Ala Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Trp Tyr

20 25 30

Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile Tyr Glu Asp Ser

35 40 45

Lys Arg Pro Ser Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly

50 55 60

Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala

65 70 75 80

Asp Tyr Tyr Cys Ser Ser Asn Thr Arg Ser Ser Thr Leu Val Phe Gly

85 90 95

Gly Gly Thr Lys Leu Thr Val Leu Gly Ser Arg Gly Gly Gly Gly Ser

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Glu Met Ala Glu Val

115 120 125

Gln Leu Val Gln Ser Gly Ala Glu Met Lys Lys Pro Gly Ala Ser Leu

130 135 140

Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ile Asp Tyr Tyr Val

145 150 155 160

Tyr Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Ser Met Gly Trp

165 170 175

Ile Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe Gln Gly

180 185 190

Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr Met Glu

195 200 205

Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr Cys Ala Arg

210 215 220

Ser Gln Arg Asp Gly Tyr Met Asp Tyr Trp Gly Gln Gly Thr Leu Val

225 230 235 240

Thr Val Ser Ser Ala Ala Ala Ile Glu Val Met Tyr Pro Pro Pro Tyr

245 250 255

Leu Asp Asn Glu Lys Ser Asn Gly Thr Ile Ile His Val Lys Gly Lys

260 265 270

His Leu Cys Pro Ser Pro Leu Phe Pro Gly Pro Ser Lys Pro Phe Trp

275 280 285

Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val

290 295 300

Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Leu

305 310 315 320

Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr

325 330 335

Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr

340 345 350

Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln

355 360 365

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

370 375 380

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

385 390 395 400

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

405 410 415

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

420 425 430

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

435 440 445

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

450 455 460

Pro Arg

465

<210> 143

<211> 473

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 143

Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly

20 25 30

Phe Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Gly Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser

85 90 95

Leu Ser Gly Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Leu Glu Met Ala Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val

130 135 140

Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr

145 150 155 160

Thr Phe Thr Asp Tyr Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln

165 170 175

Arg Leu Glu Trp Met Gly Trp Ile Asn Pro Asn Ser Gly Gly Thr Asn

180 185 190

Tyr Ala Gln Lys Phe Gln Asp Arg Ile Thr Val Thr Arg Asp Thr Ser

195 200 205

Ser Asn Thr Gly Tyr Met Glu Leu Thr Arg Leu Arg Ser Asp Asp Thr

210 215 220

Ala Val Tyr Tyr Cys Ala Arg Ser Pro Tyr Ser Gly Val Leu Asp Lys

225 230 235 240

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ala Ala Ile Glu

245 250 255

Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser Asn Gly Thr

260 265 270

Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe Pro

275 280 285

Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val Leu

290 295 300

Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val

305 310 315 320

Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr

325 330 335

Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro

340 345 350

Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser

355 360 365

Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu

370 375 380

Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg

385 390 395 400

Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln

405 410 415

Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr

420 425 430

Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp

435 440 445

Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala

450 455 460

Leu His Met Gln Ala Leu Pro Pro Arg

465 470

<210> 144

<211> 470

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 144

Ser Tyr Glu Leu Thr Gln Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Met Ser Cys Ser Gly Thr Ser Ser Asn Ile Gly Ser His

20 25 30

Ser Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Thr Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Gly Ser Leu

85 90 95

Asn Gly Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Ser

100 105 110

Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

115 120 125

Leu Glu Met Ala Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys

130 135 140

Lys Pro Gly Glu Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser

145 150 155 160

Phe Thr Ser Tyr Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly

165 170 175

Leu Glu Trp Met Gly Ile Ile Tyr Pro Gly Asp Ser Asp Thr Arg Tyr

180 185 190

Ser Pro Ser Phe Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile

195 200 205

Ser Thr Ala Tyr Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala

210 215 220

Met Tyr Tyr Cys Ala Arg Tyr Ser Gly Ser Phe Asp Asn Trp Gly Gln

225 230 235 240

Gly Thr Leu Val Thr Val Ser Ser Ala Ala Ala Ile Glu Val Met Tyr

245 250 255

Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser Asn Gly Thr Ile Ile His

260 265 270

Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe Pro Gly Pro Ser

275 280 285

Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr

290 295 300

Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys

305 310 315 320

Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg

325 330 335

Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp

340 345 350

Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala

355 360 365

Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu

370 375 380

Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp

385 390 395 400

Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu

405 410 415

Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile

420 425 430

Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr

435 440 445

Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met

450 455 460

Gln Ala Leu Pro Pro Arg

465 470

<210> 145

<211> 476

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 145

Leu Pro Val Leu Thr Gln Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ser Gly Arg Ser Ser Asn Ile Gly Ser Asn

20 25 30

Ser Val Asn Trp Tyr Arg Gln Leu Pro Gly Ala Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Ser Asn Asn Gln Arg Pro Pro Gly Val Pro Val Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Thr Tyr Tyr Cys Ala Thr Trp Asp Asp Asn Leu

85 90 95

Asn Val His Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Leu Glu Met Ala Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val

130 135 140

Lys Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly

145 150 155 160

Thr Phe Ser Ser Tyr Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln

165 170 175

Gly Leu Glu Trp Met Gly Arg Ile Ile Pro Ile Leu Gly Ile Ala Asn

180 185 190

Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser

195 200 205

Thr Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr

210 215 220

Ala Val Tyr Tyr Cys Ala Arg Gly Gly Tyr Tyr Ser His Asp Met Trp

225 230 235 240

Ser Glu Asp Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ala

245 250 255

Ala Ile Glu Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser

260 265 270

Asn Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro

275 280 285

Leu Phe Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val Gly

290 295 300

Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile

305 310 315 320

Phe Trp Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met

325 330 335

Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro

340 345 350

Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe

355 360 365

Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu

370 375 380

Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp

385 390 395 400

Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys

405 410 415

Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala

420 425 430

Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys

435 440 445

Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr

450 455 460

Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

465 470 475

<210> 146

<211> 475

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 146

Gln Ala Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn

20 25 30

Tyr Val Phe Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu

85 90 95

Ser Ala Ser Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Leu Glu Met Ala Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val

130 135 140

Lys Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly

145 150 155 160

Thr Phe Ser Ser Tyr Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln

165 170 175

Gly Leu Glu Trp Met Gly Arg Ile Ile Pro Ile Leu Gly Thr Ala Asn

180 185 190

Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser

195 200 205

Thr Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr

210 215 220

Ala Val Tyr Tyr Cys Ala Arg Ser Gly Tyr Gly Ser Tyr Arg Trp Glu

225 230 235 240

Asp Ser Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ala Ala

245 250 255

Ile Glu Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser Asn

260 265 270

Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu

275 280 285

Phe Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly

290 295 300

Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe

305 310 315 320

Trp Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn

325 330 335

Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr

340 345 350

Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser

355 360 365

Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr

370 375 380

Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys

385 390 395 400

Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn

405 410 415

Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu

420 425 430

Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly

435 440 445

His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr

450 455 460

Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

465 470 475

<210> 147

<211> 482

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 147

Leu Pro Val Leu Thr Gln Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ser Gly Arg Ser Ser Asn Ile Gly Ser Asn

20 25 30

Ser Val Asn Trp Tyr Arg Gln Leu Pro Gly Ala Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Ser Asn Asn Gln Arg Pro Pro Gly Val Pro Val Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Thr Tyr Tyr Cys Ala Thr Trp Asp Asp Asn Leu

85 90 95

Asn Val His Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Leu Glu Met Ala Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val

130 135 140

Lys Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly

145 150 155 160

Thr Phe Ser Ser Tyr Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln

165 170 175

Gly Leu Glu Trp Met Gly Arg Ile Ile Pro Ile Leu Gly Ile Ala Asn

180 185 190

Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser

195 200 205

Thr Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr

210 215 220

Ala Val Tyr Tyr Cys Ala Arg Gly Gly Tyr Tyr Ser His Asp Met Trp

225 230 235 240

Ser Glu Asp Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ala

245 250 255

Ala Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr

260 265 270

Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala

275 280 285

Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile

290 295 300

Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser

305 310 315 320

Leu Val Ile Thr Leu Tyr Cys Asn Lys Arg Gly Arg Lys Lys Leu Leu

325 330 335

Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu

340 345 350

Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys

355 360 365

Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Glu Pro Pro Ala Tyr Gln

370 375 380

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

385 390 395 400

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

405 410 415

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

420 425 430

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

435 440 445

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

450 455 460

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

465 470 475 480

Pro Arg

<210> 148

<211> 476

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 148

Ser Tyr Glu Leu Thr Gln Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Met Ser Cys Ser Gly Thr Ser Ser Asn Ile Gly Ser His

20 25 30

Ser Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Thr Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Gly Ser Leu

85 90 95

Asn Gly Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Ser

100 105 110

Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

115 120 125

Leu Glu Met Ala Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys

130 135 140

Lys Pro Gly Glu Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser

145 150 155 160

Phe Thr Ser Tyr Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly

165 170 175

Leu Glu Trp Met Gly Ile Ile Tyr Pro Gly Asp Ser Asp Thr Arg Tyr

180 185 190

Ser Pro Ser Phe Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile

195 200 205

Ser Thr Ala Tyr Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala

210 215 220

Met Tyr Tyr Cys Ala Arg Tyr Ser Gly Ser Phe Asp Asn Trp Gly Gln

225 230 235 240

Gly Thr Leu Val Thr Val Ser Ser Ala Ala Ala Pro Thr Thr Thr Pro

245 250 255

Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu

260 265 270

Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His

275 280 285

Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu

290 295 300

Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr

305 310 315 320

Cys Asn Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro

325 330 335

Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys

340 345 350

Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe

355 360 365

Ser Arg Ser Ala Glu Pro Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu

370 375 380

Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp

385 390 395 400

Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys

405 410 415

Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala

420 425 430

Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys

435 440 445

Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr

450 455 460

Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

465 470 475

<210> 149

<211> 481

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 149

Gln Ala Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn

20 25 30

Tyr Val Phe Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu

85 90 95

Ser Ala Ser Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Leu Glu Met Ala Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val

130 135 140

Lys Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly

145 150 155 160

Thr Phe Ser Ser Tyr Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln

165 170 175

Gly Leu Glu Trp Met Gly Arg Ile Ile Pro Ile Leu Gly Thr Ala Asn

180 185 190

Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser

195 200 205

Thr Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr

210 215 220

Ala Val Tyr Tyr Cys Ala Arg Ser Gly Tyr Gly Ser Tyr Arg Trp Glu

225 230 235 240

Asp Ser Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ala Ala

245 250 255

Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile

260 265 270

Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala

275 280 285

Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr

290 295 300

Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu

305 310 315 320

Val Ile Thr Leu Tyr Cys Asn Lys Arg Gly Arg Lys Lys Leu Leu Tyr

325 330 335

Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu

340 345 350

Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu

355 360 365

Leu Arg Val Lys Phe Ser Arg Ser Ala Glu Pro Pro Ala Tyr Gln Gln

370 375 380

Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu

385 390 395 400

Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly

405 410 415

Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln

420 425 430

Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu

435 440 445

Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr

450 455 460

Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro

465 470 475 480

Arg

<210> 150

<211> 479

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 150

Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly

20 25 30

Phe Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Gly Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser

85 90 95

Leu Ser Gly Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Leu Glu Met Ala Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val

130 135 140

Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr

145 150 155 160

Thr Phe Thr Asp Tyr Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln

165 170 175

Arg Leu Glu Trp Met Gly Trp Ile Asn Pro Asn Ser Gly Gly Thr Asn

180 185 190

Tyr Ala Gln Lys Phe Gln Asp Arg Ile Thr Val Thr Arg Asp Thr Ser

195 200 205

Ser Asn Thr Gly Tyr Met Glu Leu Thr Arg Leu Arg Ser Asp Asp Thr

210 215 220

Ala Val Tyr Tyr Cys Ala Arg Ser Pro Tyr Ser Gly Val Leu Asp Lys

225 230 235 240

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ala Ala Pro Thr

245 250 255

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

260 265 270

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

275 280 285

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

290 295 300

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

305 310 315 320

Thr Leu Tyr Cys Asn Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe

325 330 335

Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly

340 345 350

Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg

355 360 365

Val Lys Phe Ser Arg Ser Ala Glu Pro Pro Ala Tyr Gln Gln Gly Gln

370 375 380

Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp

385 390 395 400

Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro

405 410 415

Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp

420 425 430

Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg

435 440 445

Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr

450 455 460

Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

465 470 475

<210> 151

<211> 472

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 151

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Ala Ser Pro Gly Gln

1 5 10 15

Ser Ile Ala Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Trp Tyr

20 25 30

Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile Tyr Glu Asp Ser

35 40 45

Lys Arg Pro Ser Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly

50 55 60

Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala

65 70 75 80

Asp Tyr Tyr Cys Ser Ser Asn Thr Arg Ser Ser Thr Leu Val Phe Gly

85 90 95

Gly Gly Thr Lys Leu Thr Val Leu Gly Ser Arg Gly Gly Gly Gly Ser

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Glu Met Ala Glu Val

115 120 125

Gln Leu Val Gln Ser Gly Ala Glu Met Lys Lys Pro Gly Ala Ser Leu

130 135 140

Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ile Asp Tyr Tyr Val

145 150 155 160

Tyr Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Ser Met Gly Trp

165 170 175

Ile Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe Gln Gly

180 185 190

Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr Met Glu

195 200 205

Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr Cys Ala Arg

210 215 220

Ser Gln Arg Asp Gly Tyr Met Asp Tyr Trp Gly Gln Gly Thr Leu Val

225 230 235 240

Thr Val Ser Ser Ala Ala Ala Pro Thr Thr Thr Pro Ala Pro Arg Pro

245 250 255

Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro

260 265 270

Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu

275 280 285

Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys

290 295 300

Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Asn Lys Arg

305 310 315 320

Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro

325 330 335

Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu

340 345 350

Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala

355 360 365

Glu Pro Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu

370 375 380

Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly

385 390 395 400

Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu

405 410 415

Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser

420 425 430

Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly

435 440 445

Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu

450 455 460

His Met Gln Ala Leu Pro Pro Arg

465 470

<210> 152

<211> 472

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 152

Asp Ile Val Leu Thr Gln Ser Pro Pro Ser Leu Ala Met Ser Leu Gly

1 5 10 15

Lys Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Thr Ile Leu

20 25 30

Gly Ser His Leu Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Thr Leu Leu Ile Gln Leu Ala Ser Asn Val Gln Thr Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asp

65 70 75 80

Pro Val Glu Glu Asp Asp Val Ala Val Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Thr Ile Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys

115 120 125

Gly Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly

130 135 140

Glu Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp

145 150 155 160

Tyr Ser Ile Asn Trp Val Lys Arg Ala Pro Gly Lys Gly Leu Lys Trp

165 170 175

Met Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp

180 185 190

Phe Arg Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala

195 200 205

Tyr Leu Gln Ile Asn Asn Leu Lys Tyr Glu Asp Thr Ala Thr Tyr Phe

210 215 220

Cys Ala Leu Asp Tyr Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

225 230 235 240

Ser Val Thr Val Ser Ser Ala Ala Ala Thr Thr Thr Pro Ala Pro Arg

245 250 255

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

260 265 270

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

275 280 285

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

290 295 300

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg

305 310 315 320

Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro

325 330 335

Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu

340 345 350

Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala

355 360 365

Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu

370 375 380

Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly

385 390 395 400

Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu

405 410 415

Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser

420 425 430

Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly

435 440 445

Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu

450 455 460

His Met Gln Ala Leu Pro Pro Arg

465 470

<210> 153

<211> 472

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 153

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Ser Val Ile

20 25 30

Gly Ala His Leu Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Thr Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Ala Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Ile Phe Pro Arg Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys

115 120 125

Gly Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly

130 135 140

Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp

145 150 155 160

Tyr Ser Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp

165 170 175

Met Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp

180 185 190

Phe Arg Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala

195 200 205

Tyr Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr

210 215 220

Cys Ala Arg Asp Tyr Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

225 230 235 240

Leu Val Thr Val Ser Ser Ala Ala Ala Thr Thr Thr Pro Ala Pro Arg

245 250 255

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

260 265 270

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

275 280 285

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

290 295 300

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg

305 310 315 320

Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro

325 330 335

Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu

340 345 350

Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala

355 360 365

Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu

370 375 380

Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly

385 390 395 400

Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu

405 410 415

Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser

420 425 430

Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly

435 440 445

Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu

450 455 460

His Met Gln Ala Leu Pro Pro Arg

465 470

<210> 154

<211> 466

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 154

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Ser Val Ile

20 25 30

Gly Ala His Leu Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Thr Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Ala Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Ile Phe Pro Arg Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys

115 120 125

Gly Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly

130 135 140

Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp

145 150 155 160

Tyr Ser Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp

165 170 175

Met Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp

180 185 190

Phe Arg Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala

195 200 205

Tyr Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr

210 215 220

Cys Ala Arg Asp Tyr Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

225 230 235 240

Leu Val Thr Val Ser Ser Ala Ala Ala Asp Thr Gly Leu Tyr Ile Cys

245 250 255

Lys Val Glu Leu Met Tyr Pro Pro Pro Tyr Tyr Leu Gly Ile Gly Asn

260 265 270

Gly Thr Gln Ile Tyr Val Ile Asp Pro Glu Pro Cys Pro Asp Ser Asp

275 280 285

Phe Leu Leu Trp Ile Leu Ala Ala Val Ser Ser Gly Leu Phe Phe Tyr

290 295 300

Ser Phe Leu Leu Thr Ala Val Ser Lys Arg Gly Arg Lys Lys Leu Leu

305 310 315 320

Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu

325 330 335

Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys

340 345 350

Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln

355 360 365

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

370 375 380

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

385 390 395 400

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

405 410 415

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

420 425 430

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

435 440 445

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

450 455 460

Pro Arg

465

<210> 155

<211> 464

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 155

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Ser Val Ile

20 25 30

Gly Ala His Leu Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Thr Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Ala Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Ile Phe Pro Arg Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys

115 120 125

Gly Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly

130 135 140

Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp

145 150 155 160

Tyr Ser Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp

165 170 175

Met Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp

180 185 190

Phe Arg Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala

195 200 205

Tyr Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr

210 215 220

Cys Ala Arg Asp Tyr Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

225 230 235 240

Leu Val Thr Val Ser Ser Ala Ala Ala Gln Ile Lys Glu Ser Leu Arg

245 250 255

Ala Glu Leu Arg Val Thr Glu Arg Arg Ala Glu Val Pro Thr Ala His

260 265 270

Pro Ser Pro Ser Pro Arg Pro Ala Gly Gln Phe Gln Thr Leu Val Val

275 280 285

Gly Val Val Gly Gly Leu Leu Gly Ser Leu Val Leu Leu Val Trp Val

290 295 300

Leu Ala Val Ile Cys Ser Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile

305 310 315 320

Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp

325 330 335

Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu

340 345 350

Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly

355 360 365

Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr

370 375 380

Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys

385 390 395 400

Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys

405 410 415

Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg

420 425 430

Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala

435 440 445

Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455 460

<210> 156

<211> 653

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 156

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Val Asp Gly Asp Tyr Thr Glu Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

115 120 125

Gly Gly Gly Gly Ser Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser

130 135 140

Gly Ser Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Ser Ser Ser

145 150 155 160

Asp Val Gly Lys Tyr Asn Leu Val Ser Trp Tyr Gln Gln Pro Pro Gly

165 170 175

Lys Ala Pro Lys Leu Ile Ile Tyr Asp Val Asn Lys Arg Pro Ser Gly

180 185 190

Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Thr Leu

195 200 205

Thr Ile Ser Gly Leu Gln Gly Asp Asp Glu Ala Asp Tyr Tyr Cys Ser

210 215 220

Ser Tyr Gly Gly Ser Arg Ser Tyr Val Phe Gly Thr Gly Thr Lys Val

225 230 235 240

Thr Val Leu Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala

245 250 255

Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

260 265 270

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

275 280 285

Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

290 295 300

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

305 310 315 320

Gln Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

325 330 335

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

340 345 350

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

355 360 365

Glu Pro Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr Lys

370 375 380

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

385 390 395 400

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

405 410 415

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

420 425 430

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

435 440 445

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

450 455 460

Leu Ser Leu Ser Leu Gly Lys Met Phe Trp Val Leu Val Val Val Gly

465 470 475 480

Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile

485 490 495

Phe Trp Val Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln

500 505 510

Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser

515 520 525

Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys

530 535 540

Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln

545 550 555 560

Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu

565 570 575

Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg

580 585 590

Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met

595 600 605

Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly

610 615 620

Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp

625 630 635 640

Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

645 650

<210> 157

<211> 650

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 157

Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Gly Asp Tyr

20 25 30

Ala Met Ser Trp Phe Lys Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Phe Ile Arg Ser Lys Ala Tyr Gly Gly Thr Thr Glu Tyr Ala Ala

50 55 60

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile

65 70 75 80

Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Ala Ala Trp Ser Ala Pro Thr Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

115 120 125

Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ala Phe Leu

130 135 140

Ser Ala Ser Val Gly Asp Arg Val Thr Val Thr Cys Arg Ala Ser Gln

145 150 155 160

Gly Ile Ser Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Asn Ala

165 170 175

Pro Arg Leu Leu Ile Tyr Ser Ala Ser Thr Leu Gln Ser Gly Val Pro

180 185 190

Ser Arg Phe Arg Gly Thr Gly Tyr Gly Thr Glu Phe Ser Leu Thr Ile

195 200 205

Asp Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser

210 215 220

Tyr Thr Ser Arg Gln Thr Phe Gly Pro Gly Thr Arg Leu Asp Ile Lys

225 230 235 240

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Pro Val

245 250 255

Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

260 265 270

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

275 280 285

Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu

290 295 300

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Gln Ser Thr

305 310 315 320

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

325 330 335

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser

340 345 350

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

355 360 365

Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val

370 375 380

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

385 390 395 400

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

405 410 415

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr

420 425 430

Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val

435 440 445

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

450 455 460

Ser Leu Gly Lys Met Phe Trp Val Leu Val Val Val Gly Gly Val Leu

465 470 475 480

Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val

485 490 495

Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met

500 505 510

Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe

515 520 525

Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg

530 535 540

Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn

545 550 555 560

Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg

565 570 575

Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro

580 585 590

Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala

595 600 605

Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His

610 615 620

Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp

625 630 635 640

Ala Leu His Met Gln Ala Leu Pro Pro Arg

645 650

<210> 158

<211> 651

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 158

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Val Asp Gly Pro Ser Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Met Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

115 120 125

Gly Gly Gly Gly Ser Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser

130 135 140

Val Ala Pro Gly Gln Thr Ala Arg Ile Thr Cys Gly Ala Asn Asn Ile

145 150 155 160

Gly Ser Lys Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

165 170 175

Met Leu Val Val Tyr Asp Asp Asp Asp Arg Pro Ser Gly Ile Pro Glu

180 185 190

Arg Phe Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser

195 200 205

Gly Val Glu Ala Gly Asp Glu Ala Asp Tyr Phe Cys His Leu Trp Asp

210 215 220

Arg Ser Arg Asp His Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val

225 230 235 240

Leu Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Pro

245 250 255

Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

260 265 270

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

275 280 285

Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val

290 295 300

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Gln Ser

305 310 315 320

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

325 330 335

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser

340 345 350

Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

355 360 365

Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln

370 375 380

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

385 390 395 400

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

405 410 415

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu

420 425 430

Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser

435 440 445

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

450 455 460

Leu Ser Leu Gly Lys Met Phe Trp Val Leu Val Val Val Gly Gly Val

465 470 475 480

Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp

485 490 495

Val Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe

500 505 510

Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg

515 520 525

Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser

530 535 540

Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr

545 550 555 560

Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys

565 570 575

Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn

580 585 590

Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu

595 600 605

Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly

610 615 620

His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr

625 630 635 640

Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

645 650

<210> 159

<211> 658

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 159

Ser Tyr Glu Leu Thr Gln Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Met Ser Cys Ser Gly Thr Ser Ser Asn Ile Gly Ser His

20 25 30

Ser Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Thr Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Gly Ser Leu

85 90 95

Asn Gly Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Ser

100 105 110

Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

115 120 125

Leu Glu Met Ala Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys

130 135 140

Lys Pro Gly Glu Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser

145 150 155 160

Phe Thr Ser Tyr Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly

165 170 175

Leu Glu Trp Met Gly Ile Ile Tyr Pro Gly Asp Ser Asp Thr Arg Tyr

180 185 190

Ser Pro Ser Phe Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile

195 200 205

Ser Thr Ala Tyr Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala

210 215 220

Met Tyr Tyr Cys Ala Arg Tyr Ser Gly Ser Phe Asp Asn Trp Gly Gln

225 230 235 240

Gly Thr Leu Val Thr Val Ser Glu Ser Lys Tyr Gly Pro Pro Cys

245 250 255

Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe

260 265 270

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

275 280 285

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

290 295 300

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

305 310 315 320

Arg Glu Glu Gln Phe Gln Ser Thr Tyr Arg Val Val Ser Val Leu Thr

325 330 335

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

340 345 350

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

355 360 365

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

370 375 380

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

385 390 395 400

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

405 410 415

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

420 425 430

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

435 440 445

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

450 455 460

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Met Phe Trp Val

465 470 475 480

Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr

485 490 495

Val Ala Phe Ile Ile Phe Trp Val Lys Arg Gly Arg Lys Lys Leu Leu

500 505 510

Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu

515 520 525

Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys

530 535 540

Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln

545 550 555 560

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

565 570 575

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

580 585 590

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

595 600 605

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

610 615 620

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

625 630 635 640

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

645 650 655

Pro Arg

<210> 160

<211> 654

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 160

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Ala Ser Pro Gly Gln

1 5 10 15

Ser Ile Ala Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Trp Tyr

20 25 30

Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile Tyr Glu Asp Ser

35 40 45

Lys Arg Pro Ser Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly

50 55 60

Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala

65 70 75 80

Asp Tyr Tyr Cys Ser Ser Asn Thr Arg Ser Ser Thr Leu Val Phe Gly

85 90 95

Gly Gly Thr Lys Leu Thr Val Leu Gly Ser Arg Gly Gly Gly Gly Ser

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Glu Met Ala Glu Val

115 120 125

Gln Leu Val Gln Ser Gly Ala Glu Met Lys Lys Pro Gly Ala Ser Leu

130 135 140

Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ile Asp Tyr Tyr Val

145 150 155 160

Tyr Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Ser Met Gly Trp

165 170 175

Ile Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe Gln Gly

180 185 190

Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr Met Glu

195 200 205

Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr Cys Ala Arg

210 215 220

Ser Gln Arg Asp Gly Tyr Met Asp Tyr Trp Gly Gln Gly Thr Leu Val

225 230 235 240

Thr Val Ser Ser Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro

245 250 255

Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

260 265 270

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

275 280 285

Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val

290 295 300

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

305 310 315 320

Phe Gln Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

325 330 335

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly

340 345 350

Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

355 360 365

Arg Glu Pro Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr

370 375 380

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

385 390 395 400

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

405 410 415

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

420 425 430

Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe

435 440 445

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

450 455 460

Ser Leu Ser Leu Ser Leu Gly Lys Met Phe Trp Val Leu Val Val Val

465 470 475 480

Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile

485 490 495

Ile Phe Trp Val Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys

500 505 510

Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys

515 520 525

Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val

530 535 540

Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn

545 550 555 560

Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val

565 570 575

Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg

580 585 590

Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys

595 600 605

Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg

610 615 620

Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys

625 630 635 640

Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

645 650

<210> 161

<211> 653

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 161

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Ala Ser Pro Gly Gln

1 5 10 15

Ser Ile Ala Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Trp Tyr

20 25 30

Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile Tyr Glu Asp Ser

35 40 45

Lys Arg Pro Ser Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly

50 55 60

Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala

65 70 75 80

Asp Tyr Tyr Cys Ser Ser Asn Thr Arg Ser Ser Thr Leu Val Phe Gly

85 90 95

Gly Gly Thr Lys Leu Thr Val Leu Gly Ser Arg Gly Gly Gly Gly Ser

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Glu Met Ala Glu Val

115 120 125

Gln Leu Val Gln Ser Gly Ala Glu Met Lys Lys Pro Gly Ala Ser Leu

130 135 140

Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ile Asp Tyr Tyr Val

145 150 155 160

Tyr Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Ser Met Gly Trp

165 170 175

Ile Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe Gln Gly

180 185 190

Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr Met Glu

195 200 205

Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr Cys Ala Arg

210 215 220

Ser Gln Arg Asp Gly Tyr Met Asp Tyr Trp Gly Gln Gly Thr Leu Val

225 230 235 240

Thr Val Ser Ser Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro

245 250 255

Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

260 265 270

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

275 280 285

Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val

290 295 300

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

305 310 315 320

Phe Gln Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

325 330 335

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly

340 345 350

Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

355 360 365

Arg Glu Pro Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr

370 375 380

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

385 390 395 400

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

405 410 415

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

420 425 430

Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe

435 440 445

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

450 455 460

Ser Leu Ser Leu Ser Leu Gly Lys Met Phe Trp Val Leu Val Val Val

465 470 475 480

Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile

485 490 495

Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr

500 505 510

Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln

515 520 525

Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys

530 535 540

Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln

545 550 555 560

Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu

565 570 575

Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg

580 585 590

Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met

595 600 605

Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly

610 615 620

Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp

625 630 635 640

Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

645 650

<210> 162

<211> 462

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 162

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ala Leu Ser Asn His

20 25 30

Gly Met Ser Trp Val Arg Arg Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Val Tyr Ser Gly Ser Thr Tyr Tyr Ala Ala Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ser

85 90 95

Ala His Gly Gly Glu Ser Asp Val Trp Gly Gln Gly Thr Thr Val Thr

100 105 110

Val Ser Ser Ala Ser Gly Gly Gly Gly Ser Gly Gly Arg Ala Ser Gly

115 120 125

Gly Gly Gly Ser Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser

130 135 140

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser

145 150 155 160

Ile Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

165 170 175

Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser

180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

195 200 205

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr

210 215 220

Ser Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Thr

225 230 235 240

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

245 250 255

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

260 265 270

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

275 280 285

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

290 295 300

Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys

305 310 315 320

Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys

325 330 335

Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val

340 345 350

Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly Gln Asn

355 360 365

Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val

370 375 380

Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg

385 390 395 400

Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys

405 410 415

Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg

420 425 430

Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys

435 440 445

Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455 460

<210> 163

<211> 469

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 163

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Gly Trp Val

35 40 45

Ser Gly Ile Ser Arg Ser Gly Glu Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ser Pro Ala His Tyr Tyr Gly Gly Met Asp Val Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Arg Ala Ser Gly Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Ser

130 135 140

Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys

145 150 155 160

Arg Ala Ser Gln Ser Ile Ser Ser Ser Phe Leu Ala Trp Tyr Gln Gln

165 170 175

Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Arg Arg

180 185 190

Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp

195 200 205

Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu Asp Ser Ala Val Tyr

210 215 220

Tyr Cys Gln Gln Tyr His Ser Ser Pro Ser Trp Thr Phe Gly Gln Gly

225 230 235 240

Thr Lys Leu Glu Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr

245 250 255

Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala

260 265 270

Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe

275 280 285

Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val

290 295 300

Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys

305 310 315 320

Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr

325 330 335

Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu

340 345 350

Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro

355 360 365

Ala Tyr Lys Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly

370 375 380

Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro

385 390 395 400

Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr

405 410 415

Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly

420 425 430

Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln

435 440 445

Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln

450 455 460

Ala Leu Pro Pro Arg

465

<210> 164

<211> 462

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 164

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ala Leu Ser Asn His

20 25 30

Gly Met Ser Trp Val Arg Arg Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Val Tyr Ser Gly Ser Thr Tyr Tyr Ala Ala Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ser

85 90 95

Ala His Gly Gly Glu Ser Asp Val Trp Gly Gln Gly Thr Thr Val Thr

100 105 110

Val Ser Ser Ala Ser Gly Gly Gly Gly Ser Gly Gly Arg Ala Ser Gly

115 120 125

Gly Gly Gly Ser Asp Ile Arg Leu Thr Gln Ser Pro Ser Pro Leu Ser

130 135 140

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Glu Asp

145 150 155 160

Ile Asn Lys Phe Leu Asn Trp Tyr His Gln Thr Pro Gly Lys Ala Pro

165 170 175

Lys Leu Leu Ile Tyr Asp Ala Ser Thr Leu Gln Thr Gly Val Pro Ser

180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn

195 200 205

Ser Leu Gln Pro Glu Asp Ile Gly Thr Tyr Tyr Cys Gln Gln Tyr Glu

210 215 220

Ser Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Thr

225 230 235 240

Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser

245 250 255

Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly

260 265 270

Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp

275 280 285

Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile

290 295 300

Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys

305 310 315 320

Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys

325 330 335

Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val

340 345 350

Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly Gln Asn

355 360 365

Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val

370 375 380

Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg

385 390 395 400

Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys

405 410 415

Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg

420 425 430

Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys

435 440 445

Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455 460

<210> 165

<211> 464

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 165

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ala Leu Ser Asn His

20 25 30

Gly Met Ser Trp Val Arg Arg Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Val Tyr Ser Gly Ser Thr Tyr Tyr Ala Ala Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ser

85 90 95

Ala His Gly Gly Glu Ser Asp Val Trp Gly Gln Gly Thr Thr Val Thr

100 105 110

Val Ser Ser Ala Ser Gly Gly Gly Gly Ser Gly Gly Arg Ala Ser Gly

115 120 125

Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser

130 135 140

Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser

145 150 155 160

Ile Gly Ser Ser Ser Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala

165 170 175

Pro Arg Leu Leu Met Tyr Gly Ala Ser Ser Arg Ala Ser Gly Ile Pro

180 185 190

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

195 200 205

Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr

210 215 220

Ala Gly Ser Pro Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

225 230 235 240

Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile

245 250 255

Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala

260 265 270

Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr

275 280 285

Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu

290 295 300

Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile

305 310 315 320

Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp

325 330 335

Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu

340 345 350

Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly

355 360 365

Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr

370 375 380

Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys

385 390 395 400

Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys

405 410 415

Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg

420 425 430

Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala

435 440 445

Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

450 455 460

<210> 166

<211> 467

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 166

Gln Ile Gln Leu Val Gln Ser Gly Pro Asp Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Phe

20 25 30

Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Phe Lys Trp Met

35 40 45

Ala Trp Ile Asn Thr Tyr Thr Gly Glu Ser Tyr Phe Ala Asp Asp Phe

50 55 60

Lys Gly Arg Phe Ala Phe Ser Val Glu Thr Ser Ala Thr Thr Ala Tyr

65 70 75 80

Leu Gln Ile Asn Asn Leu Lys Thr Glu Asp Thr Ala Thr Tyr Phe Cys

85 90 95

Ala Arg Gly Glu Ile Tyr Tyr Gly Tyr Asp Gly Gly Phe Ala Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Val Val Met Thr Gln Ser

130 135 140

His Arg Phe Met Ser Thr Ser Val Gly Asp Arg Val Ser Ile Thr Cys

145 150 155 160

Arg Ala Ser Gln Asp Val Asn Thr Ala Val Ser Trp Tyr Gln Gln Lys

165 170 175

Pro Gly Gln Ser Pro Lys Leu Leu Ile Phe Ser Ala Ser Tyr Arg Tyr

180 185 190

Thr Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Ala Asp Phe

195 200 205

Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr

210 215 220

Cys Gln Gln His Tyr Ser Thr Pro Trp Thr Phe Gly Gly Gly Thr Lys

225 230 235 240

Leu Asp Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala

245 250 255

Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg

260 265 270

Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys

275 280 285

Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu

290 295 300

Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu

305 310 315 320

Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln

325 330 335

Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly

340 345 350

Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr

355 360 365

Lys Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg

370 375 380

Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met

385 390 395 400

Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu

405 410 415

Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys

420 425 430

Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu

435 440 445

Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu

450 455 460

Pro Pro Arg

465

<210> 167

<211> 472

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 167

Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Ser Ile Asn Trp Val Lys Arg Ala Pro Gly Lys Gly Leu Lys Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp Phe

50 55 60

Arg Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Asn Asn Leu Lys Tyr Glu Asp Thr Ala Thr Tyr Phe Cys

85 90 95

Ala Leu Asp Tyr Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser

100 105 110

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys

130 135 140

Lys Pro Gly Glu Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr

145 150 155 160

Phe Thr Asp Tyr Ser Ile Asn Trp Val Lys Arg Ala Pro Gly Lys Gly

165 170 175

Leu Lys Trp Met Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr

180 185 190

Ala Tyr Asp Phe Arg Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala

195 200 205

Ser Thr Ala Tyr Leu Gln Ile Asn Asn Leu Lys Tyr Glu Asp Thr Ala

210 215 220

Thr Tyr Phe Cys Ala Leu Asp Tyr Ser Tyr Ala Met Asp Tyr Trp Gly

225 230 235 240

Gln Gly Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg

245 250 255

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

260 265 270

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

275 280 285

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

290 295 300

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg

305 310 315 320

Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro

325 330 335

Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu

340 345 350

Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala

355 360 365

Asp Ala Pro Ala Tyr Lys Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu

370 375 380

Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly

385 390 395 400

Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu

405 410 415

Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser

420 425 430

Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly

435 440 445

Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu

450 455 460

His Met Gln Ala Leu Pro Pro Arg

465 470

<210> 168

<211> 466

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 168

Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Ser Ile Asn Trp Val Lys Arg Ala Pro Gly Lys Gly Leu Lys Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp Phe

50 55 60

Arg Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Asn Asn Leu Lys Tyr Glu Asp Thr Ala Thr Tyr Phe Cys

85 90 95

Ala Leu Asp Tyr Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser

100 105 110

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala

130 135 140

Met Ser Leu Gly Lys Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser

145 150 155 160

Val Ser Val Ile Gly Ala His Leu Ile His Trp Tyr Gln Gln Lys Pro

165 170 175

Gly Gln Pro Pro Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Thr

180 185 190

Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

195 200 205

Leu Thr Ile Asp Pro Val Glu Glu Asp Asp Val Ala Ile Tyr Ser Cys

210 215 220

Leu Gln Ser Arg Ile Phe Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu

225 230 235 240

Glu Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro

245 250 255

Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro

260 265 270

Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp

275 280 285

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu

290 295 300

Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu

305 310 315 320

Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu

325 330 335

Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys

340 345 350

Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys

355 360 365

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

370 375 380

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

385 390 395 400

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

405 410 415

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

420 425 430

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

435 440 445

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

450 455 460

Pro Arg

465

<210> 169

<211> 466

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 169

Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Arg His Tyr

20 25 30

Ser Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met

35 40 45

Gly Arg Ile Asn Thr Glu Ser Gly Val Pro Ile Tyr Ala Asp Asp Phe

50 55 60

Lys Gly Arg Phe Ala Phe Ser Val Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Val Ile Asn Asn Leu Lys Asp Glu Asp Thr Ala Ser Tyr Phe Cys

85 90 95

Ser Asn Asp Tyr Leu Tyr Ser Leu Asp Phe Trp Gly Gln Gly Thr Ala

100 105 110

Leu Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Ser Pro Pro Ser Leu Ala

130 135 140

Met Ser Leu Gly Lys Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser

145 150 155 160

Val Thr Ile Leu Gly Ser His Leu Ile Tyr Trp Tyr Gln Gln Lys Pro

165 170 175

Gly Gln Pro Pro Thr Leu Leu Ile Gln Leu Ala Ser Asn Val Gln Thr

180 185 190

Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr

195 200 205

Leu Thr Ile Asp Pro Val Glu Glu Asp Asp Val Ala Val Tyr Tyr Cys

210 215 220

Leu Gln Ser Arg Thr Ile Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu

225 230 235 240

Glu Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro

245 250 255

Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro

260 265 270

Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp

275 280 285

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu

290 295 300

Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu

305 310 315 320

Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu

325 330 335

Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys

340 345 350

Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys

355 360 365

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

370 375 380

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

385 390 395 400

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

405 410 415

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

420 425 430

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

435 440 445

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

450 455 460

Pro Arg

465

<210> 170

<211> 466

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 170

Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr His Tyr

20 25 30

Ser Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met

35 40 45

Gly Arg Ile Asn Thr Glu Thr Gly Glu Pro Leu Tyr Ala Asp Asp Phe

50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Val Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Phe Phe Cys

85 90 95

Ser Asn Asp Tyr Leu Tyr Ser Cys Asp Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Leu Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Ser Pro Pro Ser Leu Ala

130 135 140

Met Ser Leu Gly Lys Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser

145 150 155 160

Val Thr Ile Leu Gly Ser His Leu Ile Tyr Trp Tyr Gln Gln Lys Pro

165 170 175

Gly Gln Pro Pro Thr Leu Leu Ile Gln Leu Ala Ser Asn Val Gln Thr

180 185 190

Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr

195 200 205

Leu Thr Ile Asp Pro Val Glu Glu Asp Asp Val Ala Val Tyr Tyr Cys

210 215 220

Leu Gln Ser Arg Thr Ile Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu

225 230 235 240

Glu Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro

245 250 255

Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro

260 265 270

Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp

275 280 285

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu

290 295 300

Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu

305 310 315 320

Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu

325 330 335

Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys

340 345 350

Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys

355 360 365

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

370 375 380

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

385 390 395 400

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

405 410 415

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

420 425 430

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

435 440 445

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

450 455 460

Pro Arg

465

<210> 171

<211> 482

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 171

Asp Ile Val Leu Thr Gln Ser Pro Pro Ser Leu Ala Met Ser Leu Gly

1 5 10 15

Lys Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Thr Ile Leu

20 25 30

Gly Ser His Leu Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Thr Leu Leu Ile Gln Leu Ala Ser Asn Val Gln Thr Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asp

65 70 75 80

Pro Val Glu Glu Asp Asp Val Ala Val Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Thr Ile Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys

115 120 125

Gly Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly

130 135 140

Glu Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp

145 150 155 160

Tyr Ser Ile Asn Trp Val Lys Arg Ala Pro Gly Lys Gly Leu Lys Trp

165 170 175

Met Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp

180 185 190

Phe Arg Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala

195 200 205

Tyr Leu Gln Ile Asn Asn Leu Lys Tyr Glu Asp Thr Ala Thr Tyr Phe

210 215 220

Cys Ala Leu Asp Tyr Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

225 230 235 240

Ser Val Thr Val Ser Ser Phe Val Pro Val Phe Leu Pro Ala Lys Pro

245 250 255

Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala

260 265 270

Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly

275 280 285

Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile

290 295 300

Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val

305 310 315 320

Ile Thr Leu Tyr Cys Asn His Arg Asn Arg Ser Lys Arg Ser Arg Leu

325 330 335

Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr

340 345 350

Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr

355 360 365

Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln

370 375 380

Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu

385 390 395 400

Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly

405 410 415

Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu

420 425 430

Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly

435 440 445

Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser

450 455 460

Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro

465 470 475 480

Pro Arg

<210> 172

<211> 440

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 172

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Pro Asp Tyr

20 25 30

Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Tyr Phe Ala Ser Gly Asn Ser Glu Tyr Asn Gln Lys Phe

50 55 60

Thr Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Asn Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Ser Leu Tyr Asp Tyr Asp Trp Tyr Phe Asp Val Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr Gln Thr Pro Leu Ser

130 135 140

Leu Ser Val Thr Pro Gly Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser

145 150 155 160

Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu

165 170 175

Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn

180 185 190

Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr

195 200 205

Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Ile

210 215 220

Tyr Tyr Cys Ser Gln Ser Ser Ile Tyr Pro Trp Thr Phe Gly Gln Gly

225 230 235 240

Thr Lys Leu Glu Ile Lys Gly Leu Ala Val Ser Thr Ile Ser Ser Phe

245 250 255

Phe Pro Pro Gly Tyr Gln Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

260 265 270

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg

275 280 285

Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro

290 295 300

Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu

305 310 315 320

Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala

325 330 335

Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu

340 345 350

Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly

355 360 365

Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu

370 375 380

Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser

385 390 395 400

Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly

405 410 415

Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu

420 425 430

His Met Gln Ala Leu Pro Pro Arg

435 440

<210> 173

<211> 469

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 173

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Pro Asp Tyr

20 25 30

Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Tyr Phe Ala Ser Gly Asn Ser Glu Tyr Asn Gln Lys Phe

50 55 60

Thr Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Asn Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Ser Leu Tyr Asp Tyr Asp Trp Tyr Phe Asp Val Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr Gln Thr Pro Leu Ser

130 135 140

Leu Ser Val Thr Pro Gly Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser

145 150 155 160

Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu

165 170 175

Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn

180 185 190

Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr

195 200 205

Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Ile

210 215 220

Tyr Tyr Cys Ser Gln Ser Ser Ile Tyr Pro Trp Thr Phe Gly Gln Gly

225 230 235 240

Thr Lys Leu Glu Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr

245 250 255

Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala

260 265 270

Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe

275 280 285

Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val

290 295 300

Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys

305 310 315 320

Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr

325 330 335

Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu

340 345 350

Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro

355 360 365

Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly

370 375 380

Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro

385 390 395 400

Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr

405 410 415

Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly

420 425 430

Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln

435 440 445

Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln

450 455 460

Ala Leu Pro Pro Arg

465

<210> 174

<211> 655

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 174

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Pro Asp Tyr

20 25 30

Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Tyr Phe Ala Ser Gly Asn Ser Glu Tyr Asn Gln Lys Phe

50 55 60

Thr Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Asn Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Ser Leu Tyr Asp Tyr Asp Trp Tyr Phe Asp Val Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr Gln Thr Pro Leu Ser

130 135 140

Leu Ser Val Thr Pro Gly Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser

145 150 155 160

Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu

165 170 175

Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn

180 185 190

Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr

195 200 205

Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Ile

210 215 220

Tyr Tyr Cys Ser Gln Ser Ser Ile Tyr Pro Trp Thr Phe Gly Gln Gly

225 230 235 240

Thr Lys Leu Glu Ile Lys Glu Pro Lys Ser Pro Asp Lys Thr His Thr

245 250 255

Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu

260 265 270

Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ala Arg Thr Pro Glu

275 280 285

Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys

290 295 300

Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys

305 310 315 320

Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu

325 330 335

Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys

340 345 350

Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys

355 360 365

Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser

370 375 380

Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys

385 390 395 400

Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln

405 410 415

Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly

420 425 430

Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln

435 440 445

Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn

450 455 460

His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Tyr Ile

465 470 475 480

Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val

485 490 495

Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe

500 505 510

Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly

515 520 525

Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg

530 535 540

Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln

545 550 555 560

Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp

565 570 575

Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro

580 585 590

Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp

595 600 605

Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg

610 615 620

Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr

625 630 635 640

Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

645 650 655

<210> 175

<211> 440

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 175

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Pro Asp Tyr

20 25 30

Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Tyr Phe Ala Ser Gly Asn Ser Glu Tyr Asn Gln Lys Phe

50 55 60

Thr Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Ser Leu Tyr Asp Tyr Asp Trp Tyr Phe Asp Val Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr Gln Thr Pro Leu Ser

130 135 140

Leu Ser Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Lys Ser Ser

145 150 155 160

Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu

165 170 175

Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn

180 185 190

Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Ala

195 200 205

Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val

210 215 220

Tyr Tyr Cys Ala Glu Thr Ser His Val Pro Trp Thr Phe Gly Gln Gly

225 230 235 240

Thr Lys Leu Glu Ile Lys Gly Leu Ala Val Ser Thr Ile Ser Ser Phe

245 250 255

Phe Pro Pro Gly Tyr Gln Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

260 265 270

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg

275 280 285

Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro

290 295 300

Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu

305 310 315 320

Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala

325 330 335

Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu

340 345 350

Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly

355 360 365

Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu

370 375 380

Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser

385 390 395 400

Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly

405 410 415

Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu

420 425 430

His Met Gln Ala Leu Pro Pro Arg

435 440

<210> 176

<211> 469

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 176

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Pro Asp Tyr

20 25 30

Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Tyr Phe Ala Ser Gly Asn Ser Glu Tyr Asn Gln Lys Phe

50 55 60

Thr Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Ser Leu Tyr Asp Tyr Asp Trp Tyr Phe Asp Val Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr Gln Thr Pro Leu Ser

130 135 140

Leu Ser Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Lys Ser Ser

145 150 155 160

Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu

165 170 175

Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn

180 185 190

Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Ala

195 200 205

Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val

210 215 220

Tyr Tyr Cys Ala Glu Thr Ser His Val Pro Trp Thr Phe Gly Gln Gly

225 230 235 240

Thr Lys Leu Glu Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr

245 250 255

Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala

260 265 270

Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe

275 280 285

Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val

290 295 300

Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys

305 310 315 320

Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr

325 330 335

Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu

340 345 350

Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro

355 360 365

Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly

370 375 380

Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro

385 390 395 400

Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr

405 410 415

Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly

420 425 430

Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln

435 440 445

Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln

450 455 460

Ala Leu Pro Pro Arg

465

<210> 177

<211> 655

<212> PRT

<213> Artificial sequence

<220>

<223> CAR anti-BCMA

<400> 177

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Pro Asp Tyr

20 25 30

Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Tyr Phe Ala Ser Gly Asn Ser Glu Tyr Asn Gln Lys Phe

50 55 60

Thr Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Ser Leu Tyr Asp Tyr Asp Trp Tyr Phe Asp Val Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr Gln Thr Pro Leu Ser

130 135 140

Leu Ser Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Lys Ser Ser

145 150 155 160

Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu

165 170 175

Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn

180 185 190

Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Ala

195 200 205

Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val

210 215 220

Tyr Tyr Cys Ala Glu Thr Ser His Val Pro Trp Thr Phe Gly Gln Gly

225 230 235 240

Thr Lys Leu Glu Ile Lys Glu Pro Lys Ser Pro Asp Lys Thr His Thr

245 250 255

Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu

260 265 270

Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ala Arg Thr Pro Glu

275 280 285

Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys

290 295 300

Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys

305 310 315 320

Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu

325 330 335

Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys

340 345 350

Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys

355 360 365

Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser

370 375 380

Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys

385 390 395 400

Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln

405 410 415

Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly

420 425 430

Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln

435 440 445

Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn

450 455 460

His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Tyr Ile

465 470 475 480

Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val

485 490 495

Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe

500 505 510

Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly

515 520 525

Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg

530 535 540

Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln

545 550 555 560

Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp

565 570 575

Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro

580 585 590

Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp

595 600 605

Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg

610 615 620

Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr

625 630 635 640

Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg

645 650 655

<210> 178

<211> 28

<212> PRT

<213> Artificial sequence

<220>

<223>TM CD8a

<400> 178

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu

1 5 10 15

Ser Leu Val Ile Thr Leu Tyr Cys Asn His Arg Asn

20 25

<210> 179

<211> 21

<212> PRT

<213> Artificial sequence

<220>

<223>TM CD8a

<400> 179

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu

1 5 10 15

Ser Leu Val Ile Thr

twenty

<210> 180

<211> 4

<212> PRT

<213> Artificial sequence

<220>

<223> binding peptide

<400> 180

Arg Ala Ala Ala

one

<210> 181

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 181

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Val Asp Gly Asp Tyr Thr Glu Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 182

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 182

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Ser Ser Ser Asp Val Gly Lys Tyr

20 25 30

Asn Leu Val Ser Trp Tyr Gln Gln Pro Pro Gly Lys Ala Pro Lys Leu

35 40 45

Ile Ile Tyr Asp Val Asn Lys Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Gly Asp Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Gly Gly Ser

85 90 95

Arg Ser Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 183

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 183

Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Gly Asp Tyr

20 25 30

Ala Met Ser Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Phe Ile Arg Ser Lys Ala Tyr Gly Gly Thr Thr Glu Tyr Ala Ala

50 55 60

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile

65 70 75 80

Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Ala Ala Trp Ser Ala Pro Thr Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 184

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 184

Asp Ile Gln Met Thr Gln Ser Pro Ala Phe Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Val Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Asn Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Ser Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Arg Gly

50 55 60

Thr Gly Tyr Gly Thr Glu Phe Ser Leu Thr Ile Asp Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Thr Ser Arg Gln

85 90 95

Thr Phe Gly Pro Gly Thr Arg Leu Asp Ile Lys

100 105

<210> 185

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 185

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Val Asp Gly Pro Ser Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Met Val Thr Val Ser Ser

115

<210> 186

<211> 108

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 186

Ser Tyr Val Leu Thr Gln Pro Ser Val Ser Val Ala Pro Gly Gln

1 5 10 15

Thr Ala Arg Ile Thr Cys Gly Ala Asn Asn Ile Gly Ser Lys Ser Val

20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Met Leu Val Val Tyr

35 40 45

Asp Asp Asp Asp Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser

50 55 60

Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Val Glu Ala Gly

65 70 75 80

Asp Glu Ala Asp Tyr Phe Cys His Leu Trp Asp Arg Ser Arg Asp His

85 90 95

Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu

100 105

<210> 187

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable heavy chain

<400> 187

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Glu Asp Thr Ser Thr Asp Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ser Gly Tyr Ser Lys Ser Ile Val Ser Tyr Met Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 188

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> Anti-BCMA variable light chain

<400> 188

Leu Pro Val Leu Thr Gln Pro Ser Thr Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Val Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn

20 25 30

Val Val Phe Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Val

35 40 45

Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Val Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu

85 90 95

Ser Gly Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

<210> 189

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> Linker

<400> 189

Ala Ser Gly Gly Gly Gly Ser Gly Gly Arg Ala Ser Gly Gly Gly Gly

1 5 10 15

Ser

<---

Claims (102)

1. A method of treatment, where the method includes: (a) administering a T cell therapy to a subject having cancer, wherein the T cell therapy comprises engineered T lymphocytes expressing a chimeric antigen receptor (CAR) that specifically binds to an antigen associated with cancer; and (b) administering to the subject an immunomodulatory compound that facilitates the degradation of Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein the start of administration of said immunomodulatory compound is at least two days after the start of administration of T cell therapy. 2. A method of treatment, wherein the method comprises administering an immunomodulatory compound that facilitates the degradation of Ikaros (IKZF1) and/or Aiolos (IKZF3) to a subject having cancer, wherein: at the time of initiation of administration of the immunomodulatory compound, the subject has previously received T-cell therapy for the treatment of cancer, wherein the T-cell therapy comprises engineered T-lymphocytes expressing a chimeric antigen receptor (CAR) that specifically binds to an antigen associated with cancer; and the beginning of the introduction of the specified immunomodulatory compounds is carried out at least two days after the start of the introduction of T-cell therapy. 3. The method according to claim 1 or 2, in which the start of administration of the immunomodulatory compound is carried out after a period of time that is more than 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 24 days or 28 days after the start of T-cell therapy. 4. The method according to any one of paragraphs. 1-3, in which the cancer is untreatable or resistant to an immunomodulatory compound. 5. The method according to any one of paragraphs. 1-4, where the cancer becomes untreatable or resistant to the immunomodulatory compound after treatment with the immunomodulatory compound. 6. The method according to any one of paragraphs. 1-5, wherein the subject or cancer has a mutation or factor that renders the cancer resistant to treatment with an immunomodulatory compound. 7. The method according to any one of paragraphs. 1-6, wherein the start of administration of the immunomodulatory compound is at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the start of T cell therapy. 8. The method according to any one of paragraphs. 1-7, in which the start of administration of the immunomodulatory compound is within 96 hours, 4 days, 5 days, 6 days, 7 days, 14 days, 15 days, 21 days, 24 days, 28 days, 30 days, 36 days, 42 days, 60 days, 72 days, or 90 days after the start of T-cell therapy. 9. The method according to any one of paragraphs. 1-8, wherein the start of the administration of the immunomodulatory compound is from 2 to 28 days after the start of the administration of the T-cell therapy. 10. The method according to any one of paragraphs. 1-9, in which the start of the administration of the immunomodulatory compound is from 7 to 21 days after the start of the administration of T-cell therapy. 11. The method according to any one of paragraphs. 1-10, wherein the immunomodulatory compound is administered daily for more than 7 consecutive days, more than 14 consecutive days, more than 21 consecutive days, or more than 28 consecutive days. 12. The method according to any one of paragraphs. 1-11, wherein the immunomodulatory compound is administered daily until 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 , 27, 28, 29, 30 or more than 30 consecutive days. 13. The method according to any one of paragraphs. 1-12, in which the administration of the immunomodulatory compound continues at least from the time after the start of the administration of T-cell therapy until: the number of cells from or derived from the administered T-cell therapy detectable in the blood of the subject will not increase compared to the number of cells of the subject at the time immediately before the administration of the immunomodulatory compound or compared with the previous time after the administration of the T-cell therapy; the number of cells from or derived from T-cell therapy detectable in the blood will not be within 2.0 times (more or less) the peak or maximum number observed in the blood of the subject after the start of the introduction of T-cell therapy; the number of cells from T-cell therapy detected in the blood of the subject will not exceed 10%, 15%, 20%, 30%, 40%, 50%, or 60% of the total number of peripheral blood mononuclear cells (PBMC) in the subject's blood; the subject will not show a reduction in tumor weight compared to tumor weight at the time immediately prior to administration of the T-cell therapy or at the time immediately prior to administration of the immunomodulatory compound; and/or the subject will not demonstrate complete or clinical remission. 14. The method according to any one of paragraphs. 1-13, wherein the immunomodulatory compound is a structural analog or derivative of thalidomide. 15. The method according to any one of paragraphs. 1-14, in which the immunomodulatory compound is a compound of the following formula:

, where one of X and Y is -C(O)- and the other of X and Y is -C(O)- or -CH ₂ -; (1) R ¹ , R ² , R ³ , and R ⁴ are each independently halogen, C 1 -C 4 alkyl, or C 1 -C 4 alkoxy, or (2) one of R ¹ , R ² , R ³ and R ⁴ is -NHR ^a and the remaining R ¹ , R ² , R ³ and R ⁴ are hydrogen, where R ^a is hydrogen or alkyl from 1-8 carbon atoms; R ⁵ is hydrogen or alkyl with 1-8 carbon atoms, benzyl or halogen; provided that R ⁵ is not hydrogen, if X and Y are –C(O)–, and (i) each of R ¹ , R ² , R ³ and R ⁴ is fluoro or (ii) one of R ¹ , R ² , R ³ and R ⁴ is amino; or a pharmaceutically acceptable salt thereof. 16. The method according to any one of paragraphs. 1-15, in which the immunomodulatory compound is a compound of the following formula:

, where one of X and Y is -C(O)- and the other of X and Y is -C(O)- or -CH ₂ -, and R ⁵ is hydrogen or lower alkyl, or a pharmaceutically acceptable salt thereof. 17. The method according to any one of paragraphs. 1-14, in which the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione, pomalidomide, 3-(5- amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione, stereoisomer 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindole-2 -yl)piperidin-2,6-dione, a stereoisomer of pomalidomide, a stereoisomer of 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidin-2,6-dione or a pharmaceutically acceptable thereof salt, solvate, hydrate, co-crystal, clathrate or polymorph. 18. The method according to any one of paragraphs. 1-17, in which the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione, or its stereoisomer, or its pharmaceutically acceptable salt. 19. The method according to any one of paragraphs. 1-18, where the immunomodulatory compound is 3-(4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-2,6-dione. 20. The method according to any one of paragraphs. 1-17, where the immunomodulatory compound is pomalidomide, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. 21. The method according to any one of paragraphs. 1-17 and 20, where the immunomodulatory compound is pomalidomide. 22. The method according to any one of paragraphs. 1-14 and 17, in which the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione, or its stereoisomer, or its a pharmaceutically acceptable salt. 23. The method according to any one of paragraphs. 1-14, 17 and 22, in which the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione. 24. The method according to any one of paragraphs. 1-14, 17 and 22, wherein the immunomodulatory compound is 3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-dione hydrochloride salt. 25. The method according to any one of paragraphs. 1-14, wherein the immunomodulatory compound is 3-[4-(4-morpholin-4-ylmethylbenzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidin-2,6-dione. 26. The method according to any one of paragraphs. 1-25, wherein the immunomodulatory compound is administered orally, subcutaneously, or intravenously. 27. The method according to any one of paragraphs. 1-26 wherein the immunomodulatory compound is administered orally. 28. The method according to any one of paragraphs. 1-27, wherein the immunomodulatory compound is administered in a capsule or tablet. 29. The method according to any one of paragraphs. 1-28, wherein the immunomodulatory compound is administered at a dose of 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 0.1 mg to 10 mg, 0, 1 mg to 5 mg, 0.1 mg to 1 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 1 mg to 10 mg, 1 mg to 5 mg, 5 mg to 100 mg 5 mg to 50 mg 5 mg to 25 mg 5 mg to 10 mg 10 mg to 100 mg 10 mg to 50 mg 10 mg to 25 mg 25 mg to 100 mg, 25 mg to 50 mg, or 50 mg to 100 mg, each inclusive. 30. The method according to any one of paragraphs. 1-29, wherein the immunomodulatory compound is administered once a day, twice a day, three times a day, four times a day, five times a day, or six times a day. 31. The method according to any one of paragraphs. 1-30, wherein the immunomodulatory compound is administered at a total daily dosage amount of at least 0.1 mg per day, 0.5 mg per day, 1.0 mg per day, 2.5 mg per day, 5 mg per day, 10 mg per day, 25 mg per day, 50 mg per day, or 100 mg per day. 32. The method according to any one of paragraphs. 1–31, in which: the immunomodulatory compound is administered at a dose greater than 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg or 15 mg and less than 25 mg. 33. The method according to any one of paragraphs. 1–32, in which: the immunomodulatory compound is administered at a dose greater than 1 mg per day, 2.5 mg per day, 5 mg per day, 7.5 mg per day, 10 mg per day, or 15 mg per day and less than 25 mg per day. 34. The method according to any one of paragraphs. 1-33, in which CAR contains an extracellular domain containing an antigen-binding domain that specifically binds to an antigen. 35. The method according to any one of paragraphs. 1-34, wherein the antigen is a tumor antigen. 36. The method according to any one of paragraphs. 1-35 wherein the antigen is selected from ROR1, B-cell maturation antigen (BCMA), charcoal anhydrase 9 (CAIX), Her2/neu (erbB2 tyrosine kinase receptor), L1-CAM, CD19, CD20, CD22, mesothelin CEA and surface hepatitis B antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3 , erb-B4, erbB dimers, EGFR vIII, folate-binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, receptor containing an insert domain kinase (kdr), kappa light chain, Ley antigen, L1 cell adhesion molecule, (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, preferably expressed melanoma antigen (PRAME), survivin, TAG72, B7-H6, receptor alpha 2 IL-13 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE Al, HLA-A2 NY-ESO-1, PSCA , folate receptor-a, CD44v6, CD44v7/8, integrin avb6, 8H9, NCAM, VEGF receptors, 5T4, fetal AchR, NKG2D ligands, CD44v6, dual antigen, testicular cancer antigen, mesothelin, mouse CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY–ESO–1, MART-1, gp100, G protein-coupled 5D receptor (GPCR5D), oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c –Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms tumor 1 (WT-1), cyclin, cyclin A2, CCL-1, CD138; a pathogen-specific antigen and an antigen associated with a universal label. 37. The method according to any one of paragraphs. 1-36, in which the antigen is CD19. 38. The method according to any one of paragraphs. 1-36, wherein the antigen is BCMA. 39. The method according to any one of paragraphs. 34-38, wherein the antigen binding domain comprises an antibody or antibody fragment. 40. The method according to any one of paragraphs. 34-39, in which the antigen-binding domain is a single-stranded fragment. 41. The method of claim 39 or 40, wherein the fragment contains antibody variable regions connected by a flexible linker. 42. The method according to any one of paragraphs. 39-41, in which the fragment contains scFv. 43. The method according to any one of paragraphs. 1-42, in which CAR contains a spacer. 44. The method of claim 43 wherein the spacer is derived from an immunoglobulin. 45. The method of claim 43 wherein the spacer comprises a hinge region. 46. The method according to any one of paragraphs. 1-45, in which CAR contains an intracellular signaling region. 47. The method of claim 46, wherein the intracellular signaling region contains an intracellular signaling domain. 48. The method of claim 47 wherein the intracellular signaling domain comprises a primary signaling domain. 49. The method of claim 47 or 48, wherein the intracellular signaling domain contains a signaling domain that can induce a primary activation signal in the T-lymphocyte. 50. The method according to any one of paragraphs. 47-49, in which the intracellular signaling domain contains a signaling domain containing an immunoreceptor tyrosine activating motif (ITAM). 51. The method according to any one of paragraphs. 47-50, in which the intracellular signaling domain contains a T-lymphocyte receptor (TCR) signaling domain component. 52. The method according to any one of paragraphs. 47-51, in which the intracellular signaling domain contains an intracellular signaling domain of the CD3 chain or a signaling portion thereof. 53. The method according to any one of paragraphs. 47-52, in which the intracellular signaling domain contains the CD3-zeta (CD3ζ) chain. 54. The method according to any one of paragraphs. 46-53, wherein the recombinant receptor further comprises a transmembrane domain located between the extracellular domain and the intracellular signaling region. 55. The method of claim 54 wherein the transmembrane domain is a CD8 or CD28 transmembrane domain. 56. The method according to any one of paragraphs. 47-55, wherein the intracellular signaling region further comprises a costimulatory signaling region. 57. The method according to claim 56, wherein the costimulatory signaling region contains an intracellular signaling domain of the costimulatory T-lymphocyte molecule or a signaling portion thereof. 58. The method of claim 56 or 57, wherein the co-stimulatory signaling region comprises an intracellular signaling domain of CD28, 4-1BB or ICOS or a signaling portion thereof. 59. The method according to any one of paragraphs. 56-58, in which the costimulatory signaling region contains the 4-1BB intracellular signaling domain. 60. The method according to any one of paragraphs. 56-59, in which the costimulatory signaling region is located between the transmembrane domain and the intracellular signaling region. 61. The method according to any one of paragraphs. 1-60, wherein the T lymphocytes are selected from the group consisting of central memory T lymphocytes, effector memory T lymphocytes, intact T lymphocytes, stem central memory T lymphocytes, effector T lymphocytes, and regulatory T lymphocytes. 62. The method according to any one of paragraphs. 1–61, in which T-lymphocytes contain CD4+ T-lymphocytes. 63. The method according to any one of paragraphs. 1–61, in which T-lymphocytes contain CD8+ T-lymphocytes. 64. The method according to any one of paragraphs. 1-63, in which T-lymphocytes are primary cells obtained from the subject. 65. The method according to any one of paragraphs. 1-64, in which T-lymphocytes are autologous for the subject. 66. The method according to any one of paragraphs. 1-64, in which T-lymphocytes are allogeneic for the subject. 67. The method according to any one of paragraphs. 1–66, in which the subject is a human. 68. The method according to any one of paragraphs. 1-67, in which T-cell therapy comprises the administration of 1×10 ⁵ to 5×10 ⁸ CAR-expressing cells in total, T-lymphocytes in total, or peripheral blood mononuclear cells (PBMCs) in total, or from 1x10 ⁶ to 5x10 ⁸ CAR-expressing cells in total, T-lymphocytes in general, or peripheral blood mononuclear cells (PBMCs) in total. 69. The method according to any one of paragraphs. 1-68, wherein the T cell therapy comprises administering 1×10 ⁵ to 5×10 ⁸ CAR-expressing T lymphocytes, in total, 1×10 ⁶ to 5×10 ⁸ CAR-expressing T lymphocytes, in 2.5×10 ⁶ to 5×10 ⁸ CAR-expressing T-lymphocytes overall 5×10 ⁶ to 5×10 ⁸ CAR-expressing T-lymphocytes overall 1×10 ⁷ to 5x10 ⁸ CAR-expressing T-lymphocytes, in total 2.5x10 ⁷ to 5x10 ⁸ CAR-expressing T-lymphocytes, in total 5x10 ⁷ to 5x10 ⁸ CAR-expressing T -lymphocytes, in total, from 1x10 ⁸ to 5x10 ⁸ CAR-expressing T-lymphocytes, in general, or from 2.5x10 ⁸ to 5x10 ⁸ CAR-expressing T-lymphocytes, in general. 70. The method according to any one of paragraphs. 1-69, in which T-cell therapy includes the introduction of from 1x10 ⁵ to 1x10 ⁸ CAR-expressing cells, in general, T-lymphocytes, in general, or peripheral blood mononuclear cells (PBMC), in total, from 5 x10 ⁵ to 1 x 10 ⁷ CAR-expressing cells in total, T-lymphocytes in general, or peripheral blood mononuclear cells (PBMC) in total, or 1 x 10 ⁶ to 1 x 10 ⁷ CAR-expressing cells, in general, T-lymphocytes in general, or peripheral blood mononuclear cells (PBMC), in general, each inclusive. 71. The method according to any one of paragraphs. 1-70, in which T-cell therapy includes the introduction of no more than 1x10 ⁸ CAR-expressing cells, in total, T-lymphocytes, in general, or peripheral blood mononuclear cells (PBMC), in total, no more than 1x 10 ⁷ CAR-expressing cells, in total, T-lymphocytes, in general, or peripheral blood mononuclear cells (PBMC), in total, not more than 0.5 × 10 ⁷ CAR-expressing cells, in total, T-lymphocytes, in total, or peripheral blood mononuclear cells (PBMC), in total, not more than 1×10 ⁶ CAR-expressing cells, in total, T-lymphocytes, in total, or peripheral blood mononuclear cells (PBMC), in total, not more than 0 .5×10 ⁶ CAR-expressing cells in general, T-lymphocytes in general, or peripheral blood mononuclear cells (PBMC) in general. 72. The method according to any one of paragraphs. 1-71, in which the number of cells administered in T cell therapy is less than that in another method in which T cell therapy is administered without administration of an immunomodulatory compound. 73. The method of claim 72, wherein the number of cells administered is 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, or 10-fold less than the amount administered in the other method. 74. The method according to any one of paragraphs. 1-73, in which T-cell therapy is administered as a separate pharmaceutical composition containing T-lymphocytes. 75. The method according to any one of paragraphs. 1-73, wherein the T-cell therapy comprises a cell dose that is a divided dose, where the cells of the dose are administered in a plurality of compositions collectively containing all the cells of the dose, over a period of no more than three days. 76. The method according to any one of paragraphs. 1-75, wherein the method further comprises administering antilymphocyte chemotherapy prior to administering T cell therapy. 77. The method according to any one of paragraphs. 1-76, wherein the cancer is a B cell cancer. 78. The method according to any one of paragraphs. 1-76, wherein the cancer is myeloma, lymphoma, or leukemia. 79. The method according to any one of paragraphs. 1-78, wherein the cancer is mantle cell lymphoma (MCL), multiple myeloma (MM), acute lymphocytic leukemia (ALL), adult ALL, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL), diffuse large B-cell lymphoma (DLBCL) or follicular lymphoma (FL). 80. The method according to any one of paragraphs. 1-76, wherein the cancer is a non-hematologic cancer or is a solid tumor.

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