EP4061386A1 - Verfahren zur herstellung von multivirusspezifischen t-zellen - Google Patents
Verfahren zur herstellung von multivirusspezifischen t-zellenInfo
- Publication number
- EP4061386A1 EP4061386A1 EP19928301.1A EP19928301A EP4061386A1 EP 4061386 A1 EP4061386 A1 EP 4061386A1 EP 19928301 A EP19928301 A EP 19928301A EP 4061386 A1 EP4061386 A1 EP 4061386A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- cells
- process step
- specific
- cell
- multivirus
- 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.)
- Pending
Links
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Classifications
-
- 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
- A61K39/12—Viral antigens
-
- 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
- A61K39/464838—Viral antigens
-
- 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
-
- 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
- A61K2039/70—Multivalent vaccine
-
- 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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/10011—Adenoviridae
- C12N2710/10034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- 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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/16011—Herpesviridae
- C12N2710/16111—Cytomegalovirus, e.g. human herpesvirus 5
- C12N2710/16134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- 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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/16011—Herpesviridae
- C12N2710/16211—Lymphocryptovirus, e.g. human herpesvirus 4, Epstein-Barr Virus
- C12N2710/16234—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- 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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/22011—Polyomaviridae, e.g. polyoma, SV40, JC
- C12N2710/22034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Definitions
- the present invention relates to a method for production and expansion of particularly multivirus specific T cells which are convenient for use in antiviral treatment products and which are effective against donor-derived and multiple viruses.
- CMV cytomegalovirus
- AdV Epstein- Barr virus
- BKV BK virus
- Pharmacological agents that are used in the treatment of these infections have several side effects, especially toxicity. It is known that agents such as ganciclovir, valganciclovir, foscarnet, cidofovir, acyclovir cause side effects such as myelosuppression, leucopenia, neutropenia, nephrotoxicity, liquid-electrolyte imbalance. On the other hand, cidofovir and ribavirin agents used in AdV treatment may cause side effects in the central nervous system and toxicity; leflunomide and cidofovir agents used in BKV treatment may cause liver toxicity.
- T cells are combined with the antigen-presenting cells (APCs), which are transduced with a viral vector coding the relevant antigens or with plasmids.
- APCs antigen-presenting cells
- Antigen-presenting cells are used to stimulate T cells until the cells with enough specificity and number are reproduced.
- APCs antigen-presenting cells
- US Patent No. 6610542B1 discloses a method based on an ex-vivo expansion of CD4, CD8 and DP T cells obtained from HIV infected patients and on inhibition of the HIV gene. It is noted that the invention can be used in cell banks and organizations for restructuring immunity, to increase therapeutic effectiveness.
- virus-specific T cells T cells are incubated with multimers, which mimicking the MHC (major histocompatibility complex) bonding peptide of an APC. T cells that bonding the multimer are then isolated by using magnetic beads. Also a high amount of blood is needed to apply this method. Moreover, as the produced virus-specific T cells (VST) will be monospecific, it will not be possible to apply these to different patients.
- VST virus-specific T cells
- TR2018/05417 discloses a complex comprising an antibody- based part bonding a target antigen specifically and a virus-based peptide bonding an MHC class I protein complex. It is described that, by this complex, the virus-specific cytotoxic T cells (T memory cells or T effector cells) circulating in an individual can be directed to the said cells, by imitating an acute viral infection with MHC class I complexes of the cells expressing the target antigen to which the antibody-based part of the covalent complex specifically bonds.
- the invention provides this complex and the methods for producing and using this complex.
- T cells are activated by using necessary peptides. After the T cells are stimulated, antibodies bond to T cells secreting IFN-y by the magnetic selection, to allow their isolation. There is a need for leukapheresis in this method and the produced VSTs are monospecific like in the method of multimeric selection.
- Rapid cytotoxic T lymphocyte production is another method applied in the art.
- APC antigen-presenting cells
- Mononuclear cells are fused with peptides containing the desired viral antigens.
- APCs then promote the proliferation of T cells. Since CD45RA depletion is not performed in this method, there is a risk that naive T cells cause Graft-versus-Host disease (GVHD) in the recipient.
- T cell subgroups T S CM, T C M, T E F
- TEF effector T cell
- T S CM stem cell memory T cell
- T C M central memory T cell
- HLA human leukocyte antigen
- the present invention relates to a method for producing multivirus specific T cells, which meets the abovementioned requirements, removes all disadvantages and brings some additional advantages.
- the primary object of the invention is to develop a method for producing multivirus specific T cells that provide effective treatment against multiple viruses from donor T lymphocytes having immunity against target viruses.
- Another object of the invention is to develop a method for producing multivirus specific T cells that enable the treatment of the patient without developing resistance, without causing toxicity.
- a further object of the invention is to develop a method that provides depletion of CD45RA, which causing GVHD, and which paves the way for achieving a treatment product that is convenient for worldwide use.
- a further object of the invention is to develop a method which paves the way for a treatment product convenient for use on more than 80% of country populations, based on information obtained from HLA pools of different countries to form the necessary reaction being recognized by the viral antigen donor cells expressed by human leukocyte antigens (HLA).
- HLA human leukocyte antigens
- Yet another object of the invention is to develop a method providing stem cell memory, central memory and effector cells (T S CM, T C M, T E F) proliferation that shows longer and more effective activity against viral pathogens when compared to other T lymphocytes.
- the invention to fulfill the abovementioned objects is a method for producing multivirus specific T cells convenient for cellular therapy purposes against multiple viruses worldwide; comprising the steps of i. Making donor selections using HLA tissue information data and apheresis of blood drawn from the donor ii. Separating naive T cells (1) from apheresis allograft (2) by selectively labeling with magnetic CD45RA beads (3) iii.
- Figure 1 is a schematic view of steps (ii) to (iii) according to an embodiment of the inventive method.
- Figure 2 is a schematic view of steps (iv) to (vii) according to an embodiment of the inventive method.
- the invention is a method of production of multivirus-specific T cells convenient for use of worldwide for cellular therapy against multiple viruses; comprising the steps of i. Making donor selections using HLA tissue information data and apheresis of blood drawn from the donor ii. Separating naive T cells (1) from apheresis allograft (2) by selectively labeling with magnetic CD45RA beads (3) iii. Realizing CD45RA + depletion with the help of a magnetic column (4) and collecting naive T cells (5) under the magnetic column (4) iv. Stimulating separated mononuclear T cells (5) with a viral peptide pool (6) at least once v.
- the HLA mentioned in process step (i) is a human leukocyte antigen that forms the tissue type of the individual.
- HLA typing is used for the compatibility of patients and donors in marrow or cord blood transplants. If there is no HLA compliance during the transplant, the recipient perceives the externally given cells as foreign matter and the transplant fails from the beginning. HLA is important; because close compatibility increases the chance of transplant success, contributes to engraftment and reduces the risks associated with Graft- versus-Host Disease (GVHD).
- GVHD Graft- versus-Host Disease
- step (i) HLA pool information from different countries is examined to obtain a world-wide final product and more than one donor selection is made accordingly. At least two HLA alleles are sought for selection. In the most preferred embodiment of the invention; the compatibility of at least two HLA alleles of donors is at least 80%.
- naive T cells (1) It is known in the literature that alloreactivity is derived mainly from naive T cells (1). Therefore, in the invention, processing steps are selectively performed on naive T cells (1).
- the selective depletion of naive T cells (1) from allografts (2) protects the memory T cell response to pathogens. Therefore, in the preferred embodiment of the invention; in process step (ii), the naive T cells (1) are selectively separated from the apheresis allograft (2) from the blood obtained from the donor. Magnetic CD45RA beads (3) are used for this purpose.
- the cell suspension containing the naive T cells (1) marked with CD45RA beads (3) is passed through the magnet column (4) so that the CD45RA beads (3) remain in the magnet column (4) and CD45RA depleted mononuclear T cells (5) are collected under the magnet column (4).
- T cells collected here; T S CM, T C M, and T E F are composed of T cells.
- This step is a very important part of the invention as it eliminates the risk of compliance problems and GVHD formation in the patient the final product is transplanted.
- Graft-versus-host disease is a complex clinical syndrome resulting from severe immunological reaction mediated by healthy T lymphocytes taken from the donor and delivered to the patient with stem cells.
- the viral peptide pool (6) mentioned in process step (iv) comprises AdV, EBV, CMV, BKV peptides. These peptides are selected from the most common peptides that cause viral infection, thereby targeting to obtain a final product appealing to multiple patients.
- said viral peptide pool (6) comprises AdV5 Hexon, CMV pp65, EBV LMP2A, EBV EBNA-1 , BKV VP1 peptides.
- step (iv) is carried out on day 0 of the process.
- the specifically selected cytokine feed (7) containing IL-2 and IL-7 in process step (v) is intended to make the populations of T S CM, T C M cells survive. This step is carried out 3 times, preferably on days 0, 3 and 5 of the process; thus, virus-specific T cell activation (8) is provided.
- the viral peptide pool referred to in process step (vi) comprises AdV, EBV, CMV, BKV peptides, like process step (6).
- This step makes it possible to obtain multivirus-specific T cells (12).
- said viral peptide pool (6) comprises AdV5 Hexon, CMV pp65, EBV LMP2A, EBV EBNA-1 , BKV VP1 peptides.
- step (vi) is carried out on day 6 of the process.
- the cytokine feed (10) containing specifically selected IL-2, IL-7, and IL-15 in process step (vii) is intended to produce a cell population comprising T S CM, T C M cell populations as well as T E F cells.
- the dose of IL-2 is kept higher than the dose of IL-2 in process step (v).
- process step (vii) is carried out 3 times, on days 7, 9 and 11 of the process; thus, proliferation (11) of multivirus-specific T cells is provided.
- process step (viii) is carried out on day 13 of the process.
- VST activity and cytotoxicity experiments were performed from mononuclear cells from healthy donors.
- T2 cells B cell line
- virus-specific peptide control
- 7AAD- live T2 cell ratios were evaluated by flow cytometric analysis with CD19 labeling.
- VST activation CD25
- cytotoxicity level dead (7AAD-) CD19 + cells
- T cells With the method of the invention; without causing toxicity, it is possible to realize a product that allows the patient to be treated without developing resistance. Also, it is globally convenient as the depletion of CD45RA is provided in T cells by the method. In the method developed based on the information obtained from HLA pools, it becomes possible to obtain a treatment product convenient for use in more than 80% of the country's populations. Furthermore, compared with T lymphocytes used in the state of the art, proliferation of stem cell memory, central memory and effector T cells (T S CM, T C M, T E F), which have longer and more effective activity against viral pathogens is provided.
- T S CM, T C M, T E F central memory and effector T cells
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- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Cell Biology (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Mycology (AREA)
- Animal Behavior & Ethology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Virology (AREA)
- Hematology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
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Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| TR201918044 | 2019-11-19 | ||
| PCT/TR2019/051071 WO2021101466A1 (en) | 2019-11-19 | 2019-12-13 | A method for producing multivirus specific t cells |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| EP4061386A1 true EP4061386A1 (de) | 2022-09-28 |
| EP4061386A4 EP4061386A4 (de) | 2024-01-10 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EP19928301.1A Pending EP4061386A4 (de) | 2019-11-19 | 2019-12-13 | Verfahren zur herstellung von multivirusspezifischen t-zellen |
Country Status (2)
| Country | Link |
|---|---|
| EP (1) | EP4061386A4 (de) |
| WO (1) | WO2021101466A1 (de) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| HRP20221303T1 (hr) * | 2012-02-09 | 2022-12-23 | Baylor College Of Medicine | SMJESE PEPTIDA ZA NASTAJANJE MULTIVIRUSNIH CITOTOKSIČNIH T-LIMFOCITA(CTLs) ŠIROKE SPECIFIČNOSTI |
| KR102501827B1 (ko) * | 2016-09-16 | 2023-02-22 | 베이롤 칼리지 오브 메드신 | 바이러스-특이적인 t-세포의 활성화 및 확장을 위한 플랫폼 |
-
2019
- 2019-12-13 EP EP19928301.1A patent/EP4061386A4/de active Pending
- 2019-12-13 WO PCT/TR2019/051071 patent/WO2021101466A1/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| EP4061386A4 (de) | 2024-01-10 |
| WO2021101466A1 (en) | 2021-05-27 |
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