US20120177621A1 - Enhancement of Allogeneic Hematopoietic Stem Cell Transplantation - Google Patents

Enhancement of Allogeneic Hematopoietic Stem Cell Transplantation Download PDF

Info

Publication number
US20120177621A1
US20120177621A1 US13/347,452 US201213347452A US2012177621A1 US 20120177621 A1 US20120177621 A1 US 20120177621A1 US 201213347452 A US201213347452 A US 201213347452A US 2012177621 A1 US2012177621 A1 US 2012177621A1
Authority
US
United States
Prior art keywords
cells
memory
naive
cancer
cell
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US13/347,452
Other languages
English (en)
Inventor
Samuel Strober
Suparna Dutt
Robert Lowsky
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Leland Stanford Junior University
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to US13/347,452 priority Critical patent/US20120177621A1/en
Assigned to NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT reassignment NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT CONFIRMATORY LICENSE (SEE DOCUMENT FOR DETAILS). Assignors: THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY
Assigned to THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY reassignment THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: LOWSKY, Robert, DUTT, Suparna, STROBER, SAMUEL
Publication of US20120177621A1 publication Critical patent/US20120177621A1/en
Priority to US14/686,646 priority patent/US9504715B2/en
Priority to US15/217,363 priority patent/US9833477B2/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/461—Cellular immunotherapy characterised by the cell type used
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/461—Cellular immunotherapy characterised by the cell type used
    • A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
    • A61K39/4643—Vertebrate antigens
    • A61K39/46434—Antigens related to induction of tolerance to non-self
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/06—Antianaemics
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • C12N5/0637—Immunosuppressive T lymphocytes, e.g. regulatory T cells or Treg
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K2035/124—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells the cells being hematopoietic, bone marrow derived or blood cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
    • A61K2239/26—Universal/off- the- shelf cellular immunotherapy; Allogenic cells or means to avoid rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
    • A61K2239/38—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterised by the dose, timing or administration schedule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
    • A61K2239/46—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterised by the cancer treated
    • A61K2239/48—Blood cells, e.g. leukemia or lymphoma

Definitions

  • Cancer also known as malignant neoplasm, is characterized by an abnormal growth of cells that display uncontrolled cell division, invasion and destruction of adjacent tissues, and sometimes metastasis to other locations in the body.
  • cancer includes breast cancer, skin cancer, lung cancer, colon cancer, prostate cancer, and lymphoma. Cancer is the second leading cause of death in America and it causes about 13% of all deaths. Cancer may affect people at all ages, even fetuses, but the risk for most types of cancer increases with age. Cancers can affect all animals.
  • Chemotherapy has become the standard of care for many cancers. Chemotherapy refers to antineoplastic drugs used to treat cancer or the combination of these drugs into a cytotoxic standardized treatment regimen. Most commonly, chemotherapy acts by killing cells that divide rapidly, one of the main properties of cancer cells. This means that it also harms cells that divide rapidly under normal circumstances: cells in the bone marrow, digestive tract and hair follicles; this results in the most common side effects of chemotherapy-myelosuppression (decreased production of blood cells), mucositis (inflammation of the lining of the digestive tract) and alopecia (hair loss). Newer anticancer drugs act directly against abnormal proteins in cancer cells; this is termed targeted therapy.
  • Allogeneic hematopoietic cell transplantation can be curative for patients with leukemia and lymphomas, especially if the recipient is in complete remission at the time of transplantation.
  • HCT hematopoietic cell transplantation
  • the risk of progressive disease or of relapse is considerably greater if the recipient is in partial remission at the time of transplantation, or if mixed rather than complete chimerism develops when non-myeloablative conditioning is used.
  • compositions and methods are provided for augmenting the treatment of cancer, including without limitation leukemias and lymphomas, after allogeneic hematopoietic cell transplantation by adding a purified subset of donor lymphocytes that can kill the tumor cells without inducing the major complication of graft versus host disease (GVHD).
  • GVHD graft versus host disease
  • methods and compositions are provided to augment the conversion of mixed hematopoietic cell chimerism to complete donor cell chimerism following allogeneic hematopoietic cell transplantation (HCT), where such transplantation may be utilized for treatment of cancer or for other conditions requiring reconstitution of the hematopoietic system, e.g. treatment of anemias, thalassemias, autoimmune conditions, and the like.
  • HCT allogeneic hematopoietic cell transplantation
  • DLI donor lymphocyte infusions
  • the present invention improves on conventional DLI by utilizing a composition of substantially purified donor memory CD8 + T cells as DLI following allogeneic HCT, where the cells are administered at a suitable time following transplantation in humans, e.g. from about 2 to about 6 months to prevent tumor relapse or at the time of tumor relapse to treat the relapse.
  • the methods provide for a more complete donor chimerism, and have the further benefit of killing tumor cells.
  • the memory CD8+ T cells may include one or both of central and effector memory T cells, usually both.
  • the donor memory T cells are generally purified, and may be selected for with an affinity agent specific for CD8, alone or in combination with other markers of such cells as known in the art, including without limitation a CD8 + CD44 + population.
  • the T cells are administered in a dose effective to promote complete chimerism and, when appropriate, to enhance killing of tumor cells.
  • the cancer is a solid tumor.
  • solid tumors that can be treating using the subject methods of the present invention include but are not limited to colorectal cancer, lung cancer, breast cancer, pancreatic cancer, liver cancer, prostate cancer, and ovarian cancer.
  • the cancer is a leukemia or lymphoma, including without limitation, acute lymphocytic leukemia (ALL), acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), lymphomas such as Hodgkin and non-Hodgkin lymphomas, etc.
  • the tumor cells are a primary or metastatic tumor.
  • FIG. 1 Survival of BALB/c hosts transplanted with donor C57BL/6 TCD BM cells with or without total T or T cell subsets.
  • B Serial measurements of body weights were determined in mice from panel A. + Weight measurements were stopped when no more than 2 mice remained in the group.
  • C The hosts received 500 BCL 1 lymphoma cells 6 hours after irradiation.
  • FIG. 2 Characterization of Naive and Memory Phenotype CD8 + T cells.
  • A C57BL/6 splenocytes were stained with anti-CD8, anti-CD44, and anti-CD62L mAbs. Gated CD62L hi CD44 low and CD44 hi CD8 + T were analyzed for CCR9, a 4 ⁇ 7 , CD122, and CXCR3 expression. Gates were determined by isotype staining. Histogram plots of the respective markers are overlaid for CD44 lo CD8 + T cells (bold line) and CD44 hi CD8 + T (tinted area).
  • ⁇ indicates that 3 H-thymidine incorporation was less than 5,000 cpm
  • C Cytokine responses of C57BL/6 donor naive or memory phenotype CD8 + T cells (1 ⁇ 10 5 ) to irradiated BALB/c stimulator cells (5 ⁇ 10 5 ) in the MLR are shown at 60 h. Left panel shows the mean ⁇ SEM concentrations of IL-2; Right shows the mean ⁇ SEM concentrations of IFN- ⁇ . #, The concentration of cytokine was ⁇ 10 pg/ml. Results are representative of at least three MLR cultures. D, Sorted Naive and memory cells were used in a cytotoxicity assay against Luciferase expressing-BCL 1 cells.
  • Sorted Naive or memory phenotype CD8 + T cells were stimulated with irradiated BALB/c splenocytes for 96 hours.
  • BCL 1 lymphoma cells expressing luciferase were mixed with stimulated naive or memory phenotype cells at various effector:target ratios. Luciferase signal was measured after 16 hours. Percent cytotoxicity was then determined as compared with the same target numbers without effector cells at each time point.
  • FIG. 3 Survival and weight changes of BALB/c recipients transplanted with donor C57BL/6 TCD BM cells with or without sorted, total, naive, or memory phenotype CD8 + T cells.
  • B and D Percentage of starting body weight of host mice given TCD BM with or without sorted naive, memory, or total CD8 + T cells as in A and C. Brackets show SEs of the mean.
  • FIG. 4 Survival, weight changes and organ pathology scores of BALB/c hosts that received 500 BCL 1 lymphoma cells followed by transplantation of TCD BM from C57BL/6 donors with or without sorted naive, or memory, or total CD8 + T cells.
  • the hosts received 500 BCL 1 lymphoma cells 6 hours after irradiation.
  • B Percentage of starting body weight of host mice given TCD BM with or without sorted naive, memory, or total CD8 + T cells as in A. Brackets show SEs of the mean.
  • C Histopathologic changes induced with naive, memory phenotype, or total CD8 + T cells and TCD BM only. Representative tissue sections were obtained from hosts in A. Histopathological specimens from the liver and large intestines of hosts were obtained at 100 days after transplantation and fixed in formalin before embedding into paraffin blocks. Tissue sections of 4-5 mm thickness were stained with hematoxylin and eosin.
  • Liver portal tracts have prominent lymphocytic infiltrates compatible with grade 2 GVHD (asterix, right photo).
  • memory group third row photos
  • TCD BM with total CD8 + T cell group increased apoptosis was seen in colonic crypts (arrow) along with increased lamina limba inflammation (open arrowhead, left photo, fourth row).
  • portal inflammation and bile duct injury was seen in liver compatible with GVHD.
  • Tissue sections were stained with H&E. Each photo is representative of 5-10 hosts examined.
  • FIG. 5 Survival, chimerism, and elimination of BCL 1 tumor cells after transplantation of TCD BM with or without sorted naive or memory CD8 + T cells.
  • A Representative two-color flow cytometric analysis of CD19 versus BCL 1 -idiotype markers in the peripheral blood from recipients 28 days after total body irradiation (TBI, 800 cGy), BCL 1 cells, and a transplant of 2 ⁇ 10 6 TCD BM marrow cells with or without 0.5 million naive or memory phenotype CD8 + T cells. Untreated control recipients were given BCL 1 only. The boxes enclose BCL 1 idiotype + CD19 + cells, and percentages of cells in boxes are shown.
  • B Representative flow cytometric analysis of peripheral blood at day 28 showing percentage of donor (H-2K b+ ) cells among gated TCR ⁇ + cells.
  • FIG. 6 Comparison of trafficking and proliferation of luciferase transgenic naive and memory CD8 + T cells after transplantation with non-transgenic TCD BM.
  • BALB/c hosts were lethally irradiated, received 500 BCL 1 lymphoma cells, and then injected with 2 ⁇ 10 6 C57BL/6 (H-2 b ) wild-type TCD BM cells with 0.5 ⁇ 10 6 naive or memory phenotype CD8 + T cells from B6-L2G85 (H-2 b )luc + mice.
  • A BLI images at serial time points after transplantation.
  • B Quantitative analysis of photon emission of BLI over time. Recipients in TCD BM group died by day 28.
  • C In vivo imaging of mice, and ex vivo imaging of intestinal tract (middle position), liver (upper left position), spleen (lower right position) and lungs (upper right position) at day 3+ and day 5+ after transplantation.
  • D Lethally irradiated BALB/c recipient mice were injected with 2 ⁇ 10 6 C57BL/6 (H-2 b , Thy1.2) TCD BM cells with either CFSE-labeled 0.5 ⁇ 10 6 congenic C57BL/6 (H-2 b , Thy1.1) sorted naive or memory phenotype cells CD8 + T cells.
  • the staining intensity of CFSE from naive and memory phenotype cells Thy1.1 + in the spleen was analyzed. The shaded profile shows staining before transplantation, and the open profile shows staining after.
  • FIG. 7 Survival, weight changes, chimersim, and blood borne BCL 1 cells in recipients with progressive tumor growth treated with infusion of total T cells or memory CD8 + T cells.
  • B Survival of recipients with or without infusion.
  • C Percentage of starting body weight of host mice given TCD BM with or without infusion as in A.
  • D Upper panels shows two-color flow cytometric analysis of CD19 versus BCL 1 -idiotype markers in the peripheral blood from recipients at day 28 after transplant of 2 ⁇ 10 6 TCD BM marrow cells with (right panel) or without infusion (left panel). The boxes enclose BCL 1 idiotype + CD19 + cells. Lower panels show representative flow cytometric analysis of peripheral blood at day 28 stained for donor (H-2K b+ ) cells versus TCR ⁇ + cells among gated TCR ⁇ + cells
  • E Representative examples of BLI of lymphoma growth in mice with or without infusion.
  • purified or “to purify” refers to the removal of contaminants from a sample.
  • effector (non-memory) T cells that are potentially reactive with a host may be deleted by removal of such cells based on cell surface phenotype. The removal of undesirable cells results in an increase in the percentage of desired cells in the sample
  • the terms “treat,” “treatment,” “treating,” and the like refer to obtaining a desired pharmacologic and/or physiologic effect.
  • the effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and/or may be therapeutic in terms of a partial or complete cure for a disease and/or adverse effect attributable to the disease.
  • Treatment covers any treatment of a disease in a mammal, particularly in a human, and includes: (a) preventing the disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, e.g., causing regression of the disease, e.g., to completely or partially remove symptoms of the disease.
  • Cancer is “inhibited” if at least one symptom of the cancer is alleviated, terminated, slowed, or prevented. As used herein, cancer is also “inhibited” if recurrence or metastasis of the cancer is reduced, slowed, delayed, or prevented. Similarly, a person with cancer is “responsive” to a treatment if at least one symptom of the cancer is alleviated, terminated, slowed, or prevented. As used herein, a person with cancer is also “responsive” to a treatment if recurrence or metastasis of the cancer is reduced, slowed, delayed or prevented.
  • Memory T cell A number of phenotypically distinct memory T cells have been described in the art and phenotypically characterized. Memory T cells of interest for the present methods are generally CD8+ cells, which may include one or both, usually both, of central and effector memory cells. Central memory T cells have been described as L-selectin and CCR7 positive, as well as secreting IL-2, but not IFN ⁇ or IL-4. Effector memory T cells, are reported to lack expression of L-selectin or CCR7 but produce effector cytokines including IFN ⁇ and IL-4.
  • Such cells may be isolated from donor peripheral blood mononuclear cells, and are typically selected for expression of CD8.
  • the cells are optionally selected as being one or more of CD44 positive in mice; CD56 negative, CD57 negative, and may be subdivided by expression of CD45 markers in humans, for example CD45RO + CD45RA ⁇ .
  • CD45RO + CD45RA ⁇ See, for example, Cui et al. (2010) Immunol Rev. 236:151-66; Lefrangois et al. (2010) Immunol Rev. 235(1):206-18; Zanetti et al. (2010) Adv Exp Med Biol. 684:108-25; Suzuki et al. (2008) Hum Immunol. 69(11):751-4; Tough (2003) Trends Immunol. 24(8):404-7; each herein specifically incorporated by reference.
  • Memory T cells may be obtained from a suitable source, including human peripheral blood, bone marrow, lymph node, umbilical cord, in vitro cell cultures and the like. Such samples can be separated by centrifugation, elutriation, density gradient separation, apheresis, affinity selection, panning, FACS, centrifugation with Hypaque, etc. prior to analysis, and usually a mononuclear fraction (PBMC) will be used. Once a sample is obtained, it can be used directly, frozen, or maintained in appropriate culture medium for short periods of time. Various media can be employed to maintain cells. The samples may be obtained by any convenient procedure, such as the drawing of blood, apheresis, venipuncture, biopsy, or the like.
  • PBMC mononuclear fraction
  • An appropriate solution may be used for dispersion or suspension of the cell sample.
  • Such solution will generally be a balanced salt solution, e.g. normal saline, PBS, Hank's balanced salt solution, etc., conveniently supplemented with fetal calf serum or other naturally occurring factors, in conjunction with an acceptable buffer at low concentration, generally from 5-25 mM.
  • Convenient buffers include HEPES, phosphate buffers, lactate buffers, etc.
  • Cell staining and selection for enrichment of the desired memory T cell population will use conventional methods.
  • Techniques providing accurate selection include fluorescence activated cell sorters, magnetic selection methods, affinity columns and the like.
  • the affinity reagents may be specific receptors or ligands for the cell surface molecules indicated above.
  • these antibodies are conjugated with a label for use in separation.
  • Labels include magnetic beads, which allow for direct separation, biotin, which can be removed with avidin or streptavidin bound to a support, fluorochromes, which can be used with a fluorescence activated cell sorter, or the like, to allow for ease of separation of the particular cell type.
  • Fluorochromes that find use include phycobiliproteins, e.g. phycoerythrin and allophycocyanins, fluorescein and Texas red. Frequently each antibody is labeled with a different fluorochrome, to permit independent sorting for each marker.
  • the antibodies are added to a suspension of cells, and incubated for a period of time sufficient to bind the available cell surface antigens.
  • the incubation will usually be at least about 5 minutes and usually less than about 30 minutes. It is desirable to have a sufficient concentration of antibodies in the reaction mixture, such that the efficiency of the separation is not limited by lack of antibody.
  • the appropriate concentration is determined by titration.
  • the medium in which the cells are separated will be any medium that maintains the viability of the cells.
  • a preferred medium is phosphate buffered saline containing from 0.1 to 0.5% BSA.
  • Various media are commercially available and may be used according to the nature of the cells, including Dulbecco's Modified Eagle Medium (dMEM), Hank's Basic Salt Solution (HBSS), Dulbecco's phosphate buffered saline (dPBS), RPMI, Iscove's medium, PBS with 5 mM EDTA, etc., frequently supplemented with fetal calf serum, BSA, HSA, etc.
  • dMEM Dulbecco's Modified Eagle Medium
  • HBSS Hank's Basic Salt Solution
  • dPBS Dulbecco's phosphate buffered saline
  • RPMI Dulbecco's phosphate buffered saline
  • Iscove's medium PBS with 5 mM EDTA, etc., frequently supplemented with fetal calf serum, BSA, HSA, etc.
  • the cells are selected for the expression of cell surface markers as previously described.
  • the cells to be provided to a patient will be at least about 50% of the selected phenotype, at least about 70%, at least about 80%, at least about 90%, at least 95% or more of the desired phenotype.
  • the cells are administered in an effective dose to provide for substantially complete chimerism of the recipient hematopoietic system to the donor cell genotype. Additionally, the dose may be sufficient to inhibit or kill tumor cells, where such cells are present. As an example, for an adult human the cell dose may be at least about 10 6 cells, at least about 10 7 cells, at least about 10 8 cells, or more. For pediatric patients the dose may be reduced appropriately, e.g. at least about 5 ⁇ 10 5 , at least about 10 6 , at least about 10 7 or more cells.
  • the cells are administered to a patient following allogeneic hematopoietic cell transplantation (HCT), where such transplantation may be utilized for treatment of cancer or for other conditions requiring reconstitution of the hematopoietic system, e.g. treatment of anemias, thalassemias, autoimmune conditions, and the like, at a suitable time following transplantation in humans, e.g., from about 2 to about 6 months to prevent relapse or at the time of relapse to treat relapse.
  • HCT allogeneic hematopoietic cell transplantation
  • compositions and methods are provided for augmenting the treatment of cancer, including without limitation leukemias and lymphomas, after allogeneic hematopoietic cell transplantation by adding a purified subset of donor lymphocytes that can kill the tumor cells without inducing the major complication of graft versus host disease (GVHD).
  • GVHD graft versus host disease
  • methods and compositions are provided to augment the conversion of mixed hematopoietic cell chimerism to complete donor cell chimerism following allogeneic hematopoietic cell transplantation (HCT), where such transplantation may be utilized for treatment of cancer or for other conditions requiring reconstitution of the hematopoietic system, e.g. treatment of anemias, thalassemias, autoimmune conditions, and the like.
  • HCT allogeneic hematopoietic cell transplantation
  • DLI donor lymphocyte infusions
  • the present invention provides a method of treating cancer comprising treating a patient with a chemotherapeutic or radiological agent; substantially ablating the recipient hematopoietic system; performing allogeneic transplantation of the hematopoietic cells, including stem cells, and following the transplantation, infusing into the patient a population of donor-derived memory CD8+ T cells, as described above.
  • cancer that can be treated by the subject methods of the present invention include but are not limited to lymphomas, leukemias, and solid tumors such as colorectal cancer, lung cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, and ovarian cancer.
  • the tumor can be primary or metastatic.
  • T cell depleted bone marrow transplants were given to recipients with a B cell lymphoma after conditioning with total body irradiation. The recipients became mixed chimeras and the tumor grew progressively. The recipients were given a DLI after about 2-3 weeks that was either made up of total T cells or memory CD8+ T cells (CD8+CD44hi) purified by flow cytometry. Whereas the total T cell DLI induced lethal GVHD, the memory CD8+ DLI eradicated the tumor without causing GVHD. The CD8+ T cells also facilitated the conversion to complete donor chimerism.
  • mouse memory CD8+ T cells responded to alloantigens in vitro by proliferation and IFN-gamma secretion with little IL-2 secretion.
  • human memory CD8+ T cells responded to alloantigens in vitro with proliferation and IFN gamma secretion with little IL-2 secretion.
  • Bone marrow transplantation has become well established in the treatment of malignant disorders.
  • High-dose chemotherapy with hematopoietic stem cell support is widely used for most hematological malignancies, as well as for some solid tumors.
  • the availability of large numbers of blood stem cells, mobilized by granulocyte colony-stimulating factor and collected by leukapheresis it is possible to overcome histocompatibility barriers in HLA-mismatched patients.
  • Clinical efficacy can be measured by any method known in the art.
  • clinical efficacy of the subject treatment method is determined by measuring the clinical benefit rate (CBR).
  • the clinical benefit rate is measured by determining the sum of the percentage of patients who are in complete remission (CR), the number of patients who are in partial remission (PR) and the number of patients having stable disease (SD) at a time point at least 6 months out from the end of therapy.
  • CBR for the subject treatment method is at least about 50%. In some embodiments, CBR for the subject treatment method is at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more.
  • the preclinical data show improved tumor killing and reduced graft v host disease by treating tumor bearing hosts with allogeneic hematopoietic cell transplantation following by infusion of donor derived CD8+ memory T cells.
  • GVHD graft versus host disease
  • subject as used herein includes humans as well as other mammals.
  • treating includes achieving a therapeutic benefit and/or a prophylactic benefit.
  • therapeutic benefit is meant eradication or amelioration of the cancer.
  • a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with cancer such that an improvement is observed in the animal subject, notwithstanding the fact that the animal subject may still be afflicted with that cancer.
  • the types of cancer that can be treated using the subject methods of the present invention include but are not limited to leukemia, lymphoma, adrenal cortical cancer, anal cancer, aplastic anemia, bile duct cancer, bladder cancer, bone cancer, bone metastasis, brain cancers, central nervous system (CNS) cancers, peripheral nervous system (PNS) cancers, breast cancer, cervical cancer, childhood Non-Hodgkin's lymphoma, colon and rectum cancer, endometrial cancer, esophagus cancer, Ewing's family of tumors (e.g.
  • Ewing's sarcoma eye cancer, gallbladder cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gestational trophoblastic disease, hairy cell leukemia, Hodgkin's lymphoma, Kaposi's sarcoma, kidney cancer, laryngeal and pharyngeal cancer, acute lymphocytic leukemia, acute myeloid leukemia, children's leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, liver cancer, lung cancer, lung carcinoid tumors, male breast cancer, malignant mesothelioma, multiple myeloma, myelodysplastic syndrome, myeloproliferative disorders, nasal cavity and paranasal cancer, nasopharyngeal cancer, neuroblastoma, oral cavity and oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, pituitary tumor, prostate cancer,
  • uterine sarcoma transitional cell carcinoma
  • vaginal cancer vulvar cancer
  • mesothelioma squamous cell or epidermoid carcinoma
  • bronchial adenoma choriocarinoma
  • head and neck cancers teratocarcinoma
  • Waldenstrom's macroglobulinemia a malignant sarcoma
  • the subject method further comprises administering to a subject in need thereof an anti-tumor agent, or a pharmaceutically acceptable salt or prodrug thereof.
  • the anti-tumor agents include but are not limited to antitumor alkylating agents, antitumor antimetabolites, antitumor antibiotics, plant-derived antitumor agents, antitumor organoplatinum compounds, antitumor campthotecin derivatives, antitumor tyrosine kinase inhibitors, monoclonal antibodies, interferons, biological response modifiers, and other agents having antitumor activities, or a pharmaceutically acceptable salt thereof.
  • CD8 + CD44 hi but not CD4 + CD44 hi Memory T Cells Mediate Potent Graft Anti-Lymphoma Activity without GVHD
  • CD4 + and CD8 + naive T cells, memory CD44 hi CD4 + T cells, naive total T cells, and memory total T CD44 hi cells either induced lethal GVHD or lacked potent anti-tumor activity.
  • the tumor bearing recipients of CD8 + CD44 hi T cells had a clear survival advantage over those given CD8 + naive T cells due to the lethal GVHD induced by the latter cells.
  • the CD8+CD44 hi T cells were also used in a model of treatment of progressive lymphoma growth after bone marrow transplantation, and were able to promote complete chimerism, and eradicate the tumor without GVHD.
  • Thy1.2 mice 8 to 12 weeks old were purchased from the breeding facility of the Department of Comparative Medicine, Stanford University or The Jackson Laboratory (Bar Harbor, Me., USA).
  • the luciferase-expressing (luck) transgenic B6-L2G85 (H-2 b , Thy1.1) mice were utilized as described previously 16 . All mice were housed in a specific pathogen free facility. Care of all experimental animals was in accordance with institutional and National Institutes of Health guidelines.
  • Antibodies and Flow Cytometric Analysis FACS. Unconjugated anti-CD16/32 (2.4G2), anti-CD8 PE (53-6.7), anti-TCR ⁇ APC (H57-597), anti-CD62L FITC (Mel-14), anti-CD44 PE (IM7), anti-LPAM-1 PE ( ⁇ 4 ⁇ 7 integrin complex) (DATK32), anti-H-2K b FITC (AF6-88.5), anti-CD19 APC (1D3), anti-B220 Pacific Blue (RA36B2) monoclonal antibodies (mAbs) were purchased from BD Pharmingen (San Diego, Calif.).
  • Anti-CD8 Alexa 700 (53-6.7) and anti-Thy1.1 Pacific Blue (OX-7) were obtained from Biolegend (San Diego, Calif.).
  • Anti-CCR9 PE (242503) and anti-CXCR3 PE (220803) mAbs were purchased from R& D systems (Minneapolis, Minn.).
  • Anti-idiotype BCL 1 antibody was purified from a hybridoma secreting rat IgG2a. The antibody was conjugated with Alexa-Fluor-488 for FACS staining. Staining and flow cytometric analysis and sorting have been described in detail previously.
  • TCD BM T cell-depleted bone marrow
  • Sorted naive or memory CD8 + T cell subsets from C57BL/6 donor mice were used as responders and mixed with irradiated (3,000 cGy) allogeneic BALB/c splenocytes as stimulators in the mixed leukocyte reaction (MLR) as described.
  • MLR mixed leukocyte reaction
  • Sorted naive or memory phenotype CD8 + T cells were stimulated in a 1:2 ratio with irradiated (3,000 rads) BALB/c splenocytes for 96 hours. Cultured cells were used as effector cells, and mixed with luciferase expressing BCL 1 luc/gfp lymphoma target cells. Cytolysis was assessed by bioluminescence imaging as described before.
  • GVHD model histopathological scoring for GVHD severity, and BCL 1 tumor model.
  • Acute GVHD was induced as described previously.
  • BALB/c hosts were given 800 cGy of total body irradiation from a 200 Kv X-ray source, and injected with donor cells via the tail vein within 24 hours. Histologic assessment of liver, small bowel and colonic GVHD was performed in a blinded fashion using the histopathologic scoring system described by Kaplan et al.
  • BCL 1 is a spontaneously arising B-cell leukemia/lymphoma derived from BALB/c mice with an IgM ⁇ surface Ig phenotype. This cell line was maintained by serial passage in BALB/c mice as described previously. The use of BCL 1 tumor cells with the luc-gfp gene construct has been reported before.
  • In vivo and ex vivo bioluminescence imaging were performed according to Edinger et al. Briefly, mice were injected intraperitoneally with luciferin (10 ⁇ g/g body weight). Ten minutes later, mice were imaged using an IVIS100 charge-coupled device imaging system (Xenogen, Alameda, Calif.) for 5 minutes. Imaging data were analyzed and quantified with Living Image Software (Xenogen). Ex vivo bioluminescence imaging was performed according to the method described by Beilhack et al.
  • CFSE proliferation assay For analysis of cell proliferation, sorted naive or memory CD8 + T cells from Thy1.1 C57BL/6 donors were loaded with a Vybrant CDDA SE (carboxyfluorescein diacetate, succinimidylester) Tracer kit (Invitrogen, Carlsbad, Calif.) as described. Thy1.1 CFSE labeled naive and memory cells (0.5 ⁇ 10 6 ) along with 2 ⁇ 10 6 TCD BM cells (C57BL/6, Thy1.2) were injected into lethally irradiated BALB/c mice. On day 3+ after BMT, the CFSE staining of the infused Thy1.1 cells from the spleen were analyzed by FACS, and comparisons of the number of cell divisions were made using proliferation analysis with Flowjo software.
  • CDDA SE carboxyfluorescein diacetate, succinimidylester
  • GVHD and anti-lymphoma activity of donor T subsets were searched for freshly isolated donor T cells that had graft anti-lymphoma activity without inducing severe GVHD.
  • H-2 d lethally irradiated BALB/c
  • H-2 b donor TCD BM cells along with a constant number (0.5 ⁇ 10 6 ) of total T cells (naive and memory unseparated) or CD4 + T cells (naive and memory unseparated).
  • the CD4 + and total T cells added to TCD BM induced acute GVHD within a week of transplantation, leading to diarrhea, progressive weight loss and death of all recipients by three weeks of transplant ( FIG. 1 ).
  • Body weight and survival were significantly reduced as compared to that of recipients given TCD BM alone (p ⁇ 0.0001 and p ⁇ 0.001 respectively) ( FIGS. 1A and B).
  • CD44 hi memory CD4 + T cells do not induce lethal GVHD 10 .
  • FIGS. 1C and E The animals that received naive CD4 + T or naive total T cells along with TCD BM succumbed to lethal GVHD with severe diarrhea and weight loss within 10 days of transplantation.
  • FIGS. 1C and E The animals that received naive CD4 + T or naive total T cells along with TCD BM succumbed to lethal GVHD with severe diarrhea and weight loss within 10 days of transplantation.
  • FIGS. 1C and E The animals that received naive CD4 + T or naive total T cells along with TCD BM succumbed to lethal GVHD with severe diarrhea and weight loss within 10 days of transplantation.
  • FIGS. 1C and E The animals that received naive CD4 + T or naive total T cells along with TCD BM succumbed to lethal GVHD with severe diarrhea and weight loss within 10 days of transplantation.
  • FIGS. 1C and E The animals that received naive CD4 + T or naive total T
  • CD8 + naive and memory T cells from unimmunized donors are alloreactive.
  • Naive CD8 + T cells showed intense staining for CCR9 and dull staining for ⁇ 4 ⁇ 7 while memory CD8 + T cells showed negative staining for both ( FIG. 2A ).
  • sorted naive or memory C57BL/6 CD8 + T cells were incubated with irradiated BALB/c spleen cells as allogeneic stimulators.
  • a constant number of sorted responder cells (1 ⁇ 10 5 ) were incubated with a constant number of stimulator cells (4 ⁇ 10 5 ).
  • the proliferation was measured after 5 days in culture, and IL-2 and IFN- ⁇ cytokine secretion was measured in the supernatants after 60 h in culture.
  • Our previous studies using this culture system showed that naive CD4 + T cells were alloreactive, but that memory CD4 + T cells were not as judged by proliferation and cytokine secretion that were no greater than background.
  • FIG. 2C shows that sorted naive CD8 + T cells secreted mean concentrations of IL-2 that were 150-200 pg/ml after culture with allogeneic stimulators, and differences between the allogeneic and syngeneic cultures were significant (p ⁇ 0.01).
  • BCL 1 lymphoma target cells transduced with the luciferase gene construct.
  • the naive and memory CD8 + T were activated in allogeneic cultures with irradiated BALB/c stimulator cells for 96 hours. These activated cells were used as effector cells in different ratios against BCL 1 target cells.
  • the enzymatic activity of luciferase was used as a measure of BCL 1 cell survival.
  • the percentage of BCL 1 cell cytolysis was similar with naive and memory CD8 + T cells in all cell ratios tested ( FIG. 2D ). Therefore both the subsets displayed similar ability to kill the tumor cells.
  • FIGS. 3A and 3C show that all irradiated recipients that received only TCD BM cells survived for 100 days. Although there was a transient weight loss during the first week after irradiation, there was a recovery to baseline during the third week and stabilization thereafter ( FIG. 3B ). These mice did not show typical clinical features of GVHD including diarrhea, hunched back, ruffled fur, hair loss, and facial swelling.
  • CD8 + memory T cells mediate anti-lymphoma activity with minimal GVHD.
  • Most of the mice that received TCD BM only died by day 30 after transplantation ( FIG. 4A ) associated with the presence of BCL 1 -idiotype + tumor cells in the blood ( FIG. 5A ).
  • FIG. 4C shows representative tissue sections of the colon and the liver of the bone marrow transplant recipients.
  • the colon of recipients given TCD BM with or without memory CD8 + T cells showed preservation of crypts and goblet cells with minimal lymphocytic infiltration
  • the recipients given, naive CD8 + T cells or total CD8 + T cells showed drop out of crypts, loss of goblet cells, and considerable infiltrates.
  • the liver of recipients given TCD BM showed tumor cells surrounding blood vessels whereas recipients of naive or total CD8 + T cells showed periportal lymphocytic infiltration. Recipients of memory CD8 + T cells showed neither of the abnormalities.
  • FIG. 1 shows representative tissue sections of the colon and the liver of the bone marrow transplant recipients.
  • 4D shows the histopathology scores for GVHD lesions in liver, colon and small intestine.
  • the scores in recipients given TCD BM with or without memory CD8 + T cells p>0.05
  • naive and memory CD8 + T cells Rapid accumulation of CD8 + naive but not memory T cells in the target organs of GVHD.
  • trafficking and survival of naive and memory CD8 + T cells was evaluated by transplantation of 0.5 ⁇ 10 6 naive or memory CD8 + T cells from C57BL/6 -L2G85 luc + mice along with 2 ⁇ 10 6 TCD BM from wild type C57BL/6 donors into irradiated BALB/c recipients that received 500 BCL 1 tumor cells.
  • Naive CD8 + T cells homed to the spleen and cervical lymph nodes by day 3, and by day 5 intense signals were observed in the gastrointestinal tract and skin ( FIG. 6A ). The signals were much lower in these organs in mice that received memory CD8 + T cells ( FIG. 6A ). TCD BM controls had no signal, and these mice died by day 28 due to lymphoma as shown before. The signals from naive CD8 + T cells continued to persist in the gastrointestinal area over the entire observation period of 84 days whereas the signals were lower from memory CD8 + T cells during this period. Quantification of the photon emission by BLI demonstrated that the signals of naive CD8 + T cells increased rapidly up to day 7, and then declined to approach background by day 70 ( FIG. 6B ).
  • the signal intensity continued to be higher than the memory CD8 + T cells until day 60 (p 0.002) ( FIG. 5B ).
  • a representative proliferation assay showed that about 10% of the naive CD8 + T cells and about 5 fold more of memory CD8 + T cells had undergone 2 or fewer cell divisions as determined by the proliferation analysis using Flowjo software. These results are consistent with the MLR data ( FIG. 2B ) that shows 7 fold less 3 H-thymidine incorporation of memory versus naive CD8 + T cells after alloantigenic stimulation.
  • CD8 + memory T cells are effective treatment for progressive lymphoma after BMT.
  • Day 16 was chosen for infusion because in preliminary experiments luck tumor cells were already expanding in lymphoid tissues of the transplant recipients ( FIG. 7E ) at that time, but had not yet been detected in blood.
  • the hosts were serially monitored for the non-transduced BCL 1 tumor in blood and signs of GVHD. All hosts that did not receive the infusion died by day 35 with tumor cells in blood ( FIGS. 7B and D).
  • FIG. 7D The group given the CD8 + memory T cells showed >99% donor T cell chimerism in all hosts whereas the control group without infusion therapy showed mixed donor T cell chimersim ( FIG. 7D ).
  • FIG. 7F shows that 5 of 5 recipients given infusion therapy were complete chimeras in the T cell lineage and granulocyte/macrophage lineage whereas recipients without the infusion were all mixed chimeras in these lineages.
  • the yield of B cells was too low for accurate determination of B220 + cells.
  • the group given the total T cell infusions that developed lethal GVHD also had >99% T cell chimerism, and BCL 1 tumor cells did not appear in blood.
  • FIG. 7E shows the lymphoma growth of luciferase expressing BCL 1 cells in hosts after TCD BM transplantation with or without memory CD8 + T cell infusion therapy. There was easily detectable lymphoma accumulation in hosts without infusion at day 16, and the tumor progressed at day 30 with increased intensity and extension to additional tissues. In contrast, there was no detectable BLI signal on day 30 in mice that were infused with memory CD8 + T cells.
  • Memory total T cells lacked potent anti-tumor activity as compared to memory CD8 + T cells, since 5 fold fewer memory CD8 + T cells induced complete tumor remissions in a higher percentage of recipients than memory total T cells. Accordingly, memory CD8 + T cells became the focus of the study.
  • CD4 + total and naive T cells induce considerably more severe GVHD than CD8 + total and CD8 + naive T cells.
  • CD4 + total T cells induced uniformly lethal GVHD by about 30 days
  • CD8 + total T cells induced death in about 40% of the recipients by 75 days.
  • memory CD8 + T cells had no in vitro or in vivo alloreactivity, memory CD8 + T cells responded to allogeneic stimulator cells in the current study as judged by proliferation and cytokine secretion. However, the proliferation and IL-2 secretion of the memory cells was significantly reduced (about 7 fold) as compared to the naive cells, but IFN-gamma secretion was significantly increased. Both subsets showed potent killing of allogeneic target cells after initial alloantigenic stimulation. It is unclear why the memory CD8 + T cells were alloreactive whereas in our previous study the memory CD4 + T cells were not. The large majority of memory CD44 hi CD8 + T cells from untreated mice were central memory T cells, whereas the large majority of memory CD4 + CD44 hi T cells were effector memory T cells.
  • CD8 + CD44 hi memory T cells did not induce lethal GVHD in irradiated hosts whereas an equal number of 1 ⁇ 10 6 or 0.5 ⁇ 10 6 naive CD8 + T cells or total CD8 + T cells, induced lethal GVHD in about 40 to 50% of recipients without tumor cells.
  • Control recipients given TCD BM and BCL 1 tumor cells all died of progressive tumor growth.
  • When the latter recipients were given 0.5 ⁇ 10 6 naive or total CD8 + T cells none died from tumor growth. However, about 25% died from GVHD, and the GVHD histopathology scores in the liver, colon, and small intestine of these recipients given naive cells or total CD8 + T cells were significantly increased as compared to recipients given TCD BM cells alone.
  • Both naive and memory CD8 + T cells had the capacity to facilitate the establishment of complete chimerism and tumor eradication despite the lack of GVHD by the memory cells.
  • the increased potency of GVHD of the naive as compared to memory cells was associated with more rapid accumulation and expansion in the lymphoid tissues, liver, and intestines as judged by bioluminescence imaging and CFSE staining.
  • the increased IL-2 secretion of the naive T cells may contribute to the increased early proliferation, and the increased expression of CCR9 and ⁇ 4 ⁇ 7 integrin gut homing receptors may contribute to the increased naive T cell early trafficking to the intestines.
  • CD8 + T cells it is likely that the ability of memory CD8 + T cells to eradicate the tumor cells is dependent on their alloreactivity, since C57BL/6 CD8 + T cells tolerized to BALB/c alloantigens lose their graft anti-tumor activity against BCL 1 lymphoma. Alloreactivity of memory CD8 + T cells can be explained by cross reactivity with viral antigens in the environment that have been shown to enhance immune responses to alloantigens on organ transplants. In addition naive CD8 + T cells can masquerade as memory phenotype cells after homeostatic proliferation with maintenance of the naive T cell TCR repertoire.
  • the current study also investigated CD8 + CD44 hi memory and unseparated total T cells as posttransplant infusion therapy in recipients given TCD BM cells and tumor cells at day 0.
  • the infusion was administered at day 16, a time point at which tumor expansion in the lymphoid tissues was apparent by bioluminescence imaging.
  • unseparated total T cells were used for infusion, then acute lethal GVHD was observed in all recipients.
  • purified CD8 + memory T cells allowed for the survival of 100% of the recipients, and survivors remained free of tumor for at least 100 days.
  • the controls given TCD bone marrow without the infusion all succumbed to tumor growth.
  • CD8 + memory T cells containing both central and effector memory subsets were able to separate GVHD and anti-lymphoma activity when added to TCD BM transplants.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Biomedical Technology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Hematology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Epidemiology (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Developmental Biology & Embryology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Mycology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Virology (AREA)
  • Oncology (AREA)
  • Diabetes (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
US13/347,452 2011-01-10 2012-01-10 Enhancement of Allogeneic Hematopoietic Stem Cell Transplantation Abandoned US20120177621A1 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
US13/347,452 US20120177621A1 (en) 2011-01-10 2012-01-10 Enhancement of Allogeneic Hematopoietic Stem Cell Transplantation
US14/686,646 US9504715B2 (en) 2011-01-10 2015-04-14 Enhancement of allogeneic hematopoietic stem cell transplantation
US15/217,363 US9833477B2 (en) 2011-01-10 2016-07-22 Enhancement of allogeneic hematopoietic stem cell transplantation

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201161460981P 2011-01-10 2011-01-10
US13/347,452 US20120177621A1 (en) 2011-01-10 2012-01-10 Enhancement of Allogeneic Hematopoietic Stem Cell Transplantation

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US14/686,646 Continuation US9504715B2 (en) 2011-01-10 2015-04-14 Enhancement of allogeneic hematopoietic stem cell transplantation

Publications (1)

Publication Number Publication Date
US20120177621A1 true US20120177621A1 (en) 2012-07-12

Family

ID=46455419

Family Applications (3)

Application Number Title Priority Date Filing Date
US13/347,452 Abandoned US20120177621A1 (en) 2011-01-10 2012-01-10 Enhancement of Allogeneic Hematopoietic Stem Cell Transplantation
US14/686,646 Active US9504715B2 (en) 2011-01-10 2015-04-14 Enhancement of allogeneic hematopoietic stem cell transplantation
US15/217,363 Active US9833477B2 (en) 2011-01-10 2016-07-22 Enhancement of allogeneic hematopoietic stem cell transplantation

Family Applications After (2)

Application Number Title Priority Date Filing Date
US14/686,646 Active US9504715B2 (en) 2011-01-10 2015-04-14 Enhancement of allogeneic hematopoietic stem cell transplantation
US15/217,363 Active US9833477B2 (en) 2011-01-10 2016-07-22 Enhancement of allogeneic hematopoietic stem cell transplantation

Country Status (8)

Country Link
US (3) US20120177621A1 (fr)
EP (1) EP2663332B1 (fr)
JP (1) JP6096677B2 (fr)
CN (2) CN104906139B (fr)
AU (1) AU2012205643A1 (fr)
CA (1) CA2823782C (fr)
GB (1) GB2500161B (fr)
WO (1) WO2012096974A1 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014133729A1 (fr) * 2013-02-26 2014-09-04 The Board Of Trustees Of The Leland Stanford Junior University Organe et cellules hématopoïétiques combinés pour tolérance de transplantation de greffes
US10300090B2 (en) 2017-03-15 2019-05-28 Orca Biosystems, Inc. Compositions of hematopoietic stem cell transplants
US10842821B2 (en) 2018-04-05 2020-11-24 Medeor Therapeutics, Inc. Cellular compositions derived from prior organ donors and methods of manufacture and use thereof
US10881692B2 (en) 2018-04-05 2021-01-05 Medeor Therapeutics, Inc. Compositions for establishing mixed chimerism and methods of manufacture thereof
US11273179B2 (en) 2018-03-12 2022-03-15 Medeor Therapeutics, Inc. Methods for treating non-cancerous disorders using hematopoietic cells
US11435350B2 (en) 2018-09-18 2022-09-06 Medeor Therapeutics, Inc. Methods of analysis of blood from deceased donors
US11813376B2 (en) 2018-09-18 2023-11-14 Medeor Therapeutics, Inc. Cellular compositions derived from deceased donors to promote graft tolerance and manufacture and uses thereof

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014172532A2 (fr) 2013-04-17 2014-10-23 Lowsky Robert Procédés permettant un traitement du cancer
US20200108098A1 (en) * 2018-02-23 2020-04-09 H. Lee Moffitt Cancer Center And Research Institute, Inc. Anti-cd83 chimeric antigen receptor expressing t regulatory cells

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0831860B1 (fr) * 1995-05-25 2004-10-20 Baxter International Inc. Therapie cellulaire allogenique anticancereuse suivant une transplantation de cellules souches allogeniques
DE69916302T2 (de) * 1998-11-11 2005-05-04 Emory University Verwendung von dendritischen zellen-depletierten zusammensetzungen zur verbesserung der transplantation von allogenen hämatopoietischen zellen
US7435592B2 (en) * 2003-05-13 2008-10-14 Immunovative Therapies, Ltd. Compositions for allogeneic cell therapy
WO2012032526A2 (fr) * 2010-09-08 2012-03-15 Yeda Research And Development Co. Ltd. Utilisation de lymphocytes t de mémoire centrale anti-tiers dans le traitement contre la leucémie et le lymphome
RU2506311C2 (ru) 2008-10-30 2014-02-10 Йеда Рисёрч Энд Девелопмент Ко. Лтд. Т-клетки центральной памяти против третьей стороны, способы их получения и их применение в трансплантации и лечении заболеваний

Non-Patent Citations (11)

* Cited by examiner, † Cited by third party
Title
Chen et al., Blood, Vol. 109, No. 7, pp. 3115-3123 (2007). *
Dutt et al., ASH poster and abstract (2009) *
Liang et al., Blood, 105(5):2180-2188 (2005) *
Lichtman et al., J. Immunol., 158: 3640-3650 (1997). *
Machura et al., Arch. Immunol. Ther. Exp., 56:55-62 (2008) *
Maldonado et al (Arthritis Res. Ther., Vol. 5., No. 2, R91-R96 (2003). *
Riddell et al., PLoS Medic., Vol. 4, Issue 7, pp. 1174-1177 (2007). *
Zhang et al., Blood, Vol. 103, No. 10, pp. 3970-3978 (2004) *
Zhang et al., J. Immunol., 178:838-850 (2007) *
Zhang et al., Nature Medic., Vol. 11, No. 12 1299-1305 (2005). *
Zheng et al., J. Immunol., 1778:838-850 (2007). *

Cited By (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11129849B2 (en) 2013-02-26 2021-09-28 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US9504717B2 (en) 2013-02-26 2016-11-29 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of HLA mismatched grafts
CN105188755A (zh) * 2013-02-26 2015-12-23 小利兰·斯坦福大学托管委员会 实现移植物移植耐受的联合器官和造血细胞
US11648264B2 (en) 2013-02-26 2023-05-16 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of HLA mismatched grafts
US9561253B2 (en) 2013-02-26 2017-02-07 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US11458165B2 (en) 2013-02-26 2022-10-04 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US9974807B2 (en) 2013-02-26 2018-05-22 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US10076542B2 (en) 2013-02-26 2018-09-18 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US10080769B2 (en) 2013-02-26 2018-09-25 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of HLA mismatched grafts
US10159694B2 (en) 2013-02-26 2018-12-25 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US10183043B2 (en) 2013-02-26 2019-01-22 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US10258648B2 (en) 2013-02-26 2019-04-16 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US11439662B2 (en) 2013-02-26 2022-09-13 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US10555971B2 (en) 2013-02-26 2020-02-11 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of HLA mismatched grafts
US10603340B2 (en) 2013-02-26 2020-03-31 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
AU2018250369B2 (en) * 2013-02-26 2020-09-03 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US11426429B2 (en) 2013-02-26 2022-08-30 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
GB2526228A (en) * 2013-02-26 2015-11-18 Univ Leland Stanford Junior Combined organ and hematopoietic cells for transplantation tolerance of grafts
GB2526228B (en) * 2013-02-26 2017-05-03 Univ Leland Stanford Junior Combined organ and hematopoietic cells for transplantation tolerance of grafts
US11007220B2 (en) 2013-02-26 2021-05-18 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US11020428B2 (en) 2013-02-26 2021-06-01 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US11040067B2 (en) 2013-02-26 2021-06-22 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US11116794B2 (en) 2013-02-26 2021-09-14 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
US11123367B2 (en) 2013-02-26 2021-09-21 The Board Of Trustees Of The Leland Stanford Junior University Combined organ and hematopoietic cells for transplantation tolerance of grafts
WO2014133729A1 (fr) * 2013-02-26 2014-09-04 The Board Of Trustees Of The Leland Stanford Junior University Organe et cellules hématopoïétiques combinés pour tolérance de transplantation de greffes
US10300090B2 (en) 2017-03-15 2019-05-28 Orca Biosystems, Inc. Compositions of hematopoietic stem cell transplants
US10857183B2 (en) 2017-03-15 2020-12-08 Orca Biosystems, Inc. Method of hematopoietic stem cell transplants
US12011461B2 (en) 2017-03-15 2024-06-18 Orca Biosystems, Inc. Compositions and methods of hematopoietic stem cell transplants
US11273179B2 (en) 2018-03-12 2022-03-15 Medeor Therapeutics, Inc. Methods for treating non-cancerous disorders using hematopoietic cells
US11819521B2 (en) 2018-04-05 2023-11-21 Medeor Therapeutics, Inc. Cellular compositions derived from prior organ donors and methods of manufacture and use thereof
US10881692B2 (en) 2018-04-05 2021-01-05 Medeor Therapeutics, Inc. Compositions for establishing mixed chimerism and methods of manufacture thereof
US11701392B2 (en) 2018-04-05 2023-07-18 Medeor Therapeutics, Inc. Compositions for establishing mixed chimerism and methods of manufacture thereof
US10842821B2 (en) 2018-04-05 2020-11-24 Medeor Therapeutics, Inc. Cellular compositions derived from prior organ donors and methods of manufacture and use thereof
US11435350B2 (en) 2018-09-18 2022-09-06 Medeor Therapeutics, Inc. Methods of analysis of blood from deceased donors
US11813376B2 (en) 2018-09-18 2023-11-14 Medeor Therapeutics, Inc. Cellular compositions derived from deceased donors to promote graft tolerance and manufacture and uses thereof

Also Published As

Publication number Publication date
JP6096677B2 (ja) 2017-03-15
EP2663332A1 (fr) 2013-11-20
JP2014502637A (ja) 2014-02-03
US9833477B2 (en) 2017-12-05
CA2823782C (fr) 2019-05-07
US9504715B2 (en) 2016-11-29
WO2012096974A1 (fr) 2012-07-19
EP2663332A4 (fr) 2014-07-02
CN104906139A (zh) 2015-09-16
US20150216900A1 (en) 2015-08-06
CA2823782A1 (fr) 2012-07-19
AU2012205643A1 (en) 2013-07-25
EP2663332B1 (fr) 2017-10-11
CN104906139B (zh) 2020-10-23
CN103402541A (zh) 2013-11-20
GB2500161B (en) 2018-12-26
US20170014453A1 (en) 2017-01-19
GB2500161A (en) 2013-09-11
GB201312433D0 (en) 2013-08-28

Similar Documents

Publication Publication Date Title
US9833477B2 (en) Enhancement of allogeneic hematopoietic stem cell transplantation
Mo et al. Engineered off-the-shelf therapeutic T cells resist host immune rejection
US20210228632A1 (en) Compostion for use in immunotherapy
Lamb et al. Natural killer cell therapy for hematologic malignancies: successes, challenges, and the future
Dutt et al. CD8+ CD44hi but not CD4+ CD44hi memory T cells mediate potent graft antilymphoma activity without GVHD
Flutter et al. Nonhematopoietic antigen blocks memory programming of alloreactive CD8+ T cells and drives their eventual exhaustion in mouse models of bone marrow transplantation
Kroesen et al. A transplantable TH‐MYCN transgenic tumor model in C57Bl/6 mice for preclinical immunological studies in neuroblastoma
Simonetta et al. Allogeneic CAR invariant natural killer T cells exert potent antitumor effects through host CD8 T-cell cross-priming
Zaghi et al. Innate immune responses in the outcome of haploidentical hematopoietic stem cell transplantation to cure hematologic malignancies
Andrlová et al. An unconventional view of T cell reconstitution after allogeneic hematopoietic cell transplantation
Jiang et al. The contribution of cytotoxic and noncytotoxic function by donor T-cells that support engraftment after allogeneic bone marrow transplantation
US10201572B2 (en) Combination therapy with double negative T-cells
US20240075063A1 (en) Use of mait cells for controlling graft versus host disease
AU2015268727B2 (en) Enhancement of allogeneic hematopoietic stem cell transplantation
Ophir et al. Induction of tolerance in organ recipients by hematopoietic stem cell transplantation
Oh et al. T Cell–Mediated Rejection of Human CD34+ Cells Is Prevented by Costimulatory Blockade in a Xenograft Model
Koporc et al. Induction of mixed chimerism through transplantation of CD45-congenic mobilized peripheral blood stem cells after nonmyeloablative irradiation
Morecki et al. Induction of graft-versus-leukemia (GVL) effect without graft-versus-host disease (GVHD) by pretransplant donor treatment with immunomodulators
Soh et al. Optimizing Blood Stem Cell Transplants Through Cellular Engineering
Ophir et al. Induction of transplantation tolerance in haploidenical transplantation under reduced intensity conditioning: the role of ex-vivo generated donor CD8+ T cells with central memory phenotype
Galaverna Outcome and immune reconstitution of children and young adults with acute leukemia after alfa/beta T-cell and B-cell depleted HLA-Haploidentical transplantation
Or-Geva induction of immune tolerance by veto cells
Park et al. Bone marrow T cells are superior to splenic T cells to induce chimeric conversion after non-myeloablative bone marrow transplantation
Lu et al. Molecular Chimeric Recipient Precursor T Cells Promote Cardiac Allograft Survival in Mice
SUPPORTIVE Cord blood oral abstracts

Legal Events

Date Code Title Description
AS Assignment

Owner name: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF

Free format text: CONFIRMATORY LICENSE;ASSIGNOR:THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY;REEL/FRAME:027621/0575

Effective date: 20120130

AS Assignment

Owner name: THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIO

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:STROBER, SAMUEL;DUTT, SUPARNA;LOWSKY, ROBERT;SIGNING DATES FROM 20120202 TO 20120207;REEL/FRAME:027799/0263

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION