WO2021101466A1 - A method for producing multivirus specific t cells - Google Patents
A method for producing multivirus specific t cells Download PDFInfo
- Publication number
- WO2021101466A1 WO2021101466A1 PCT/TR2019/051071 TR2019051071W WO2021101466A1 WO 2021101466 A1 WO2021101466 A1 WO 2021101466A1 TR 2019051071 W TR2019051071 W TR 2019051071W WO 2021101466 A1 WO2021101466 A1 WO 2021101466A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cells
- process step
- specific
- cell
- multivirus
- Prior art date
Links
- 210000001744 T-lymphocyte Anatomy 0.000 title claims abstract description 73
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 16
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 32
- 241000700605 Viruses Species 0.000 claims abstract description 24
- 210000004027 cell Anatomy 0.000 claims abstract description 23
- 230000003612 virological effect Effects 0.000 claims abstract description 19
- 108010002350 Interleukin-2 Proteins 0.000 claims abstract description 18
- 108090000695 Cytokines Proteins 0.000 claims abstract description 14
- 102000004127 Cytokines Human genes 0.000 claims abstract description 14
- 108010002586 Interleukin-7 Proteins 0.000 claims abstract description 14
- 230000000694 effects Effects 0.000 claims abstract description 13
- 238000002617 apheresis Methods 0.000 claims abstract description 9
- 239000011324 bead Substances 0.000 claims abstract description 9
- 108090000172 Interleukin-15 Proteins 0.000 claims abstract description 7
- 210000004369 blood Anatomy 0.000 claims abstract description 7
- 239000008280 blood Substances 0.000 claims abstract description 7
- 230000029036 donor selection Effects 0.000 claims abstract description 6
- 238000002659 cell therapy Methods 0.000 claims abstract description 5
- 238000002372 labelling Methods 0.000 claims abstract description 5
- 230000004936 stimulating effect Effects 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims description 73
- 230000008569 process Effects 0.000 claims description 10
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 9
- 108700028369 Alleles Proteins 0.000 claims description 4
- 108091008048 CMVpp65 Proteins 0.000 claims description 4
- 108010031111 EBV-encoded nuclear antigen 1 Proteins 0.000 claims description 4
- NTIZESTWPVYFNL-UHFFFAOYSA-N Methyl isobutyl ketone Chemical compound CC(C)CC(C)=O NTIZESTWPVYFNL-UHFFFAOYSA-N 0.000 claims description 4
- 108010023958 glycyl-threonyl-alanyl-methionyl-arginyl-isoleucyl-leucyl-glycyl-glycyl-valyl-isoleucine Proteins 0.000 claims description 4
- 238000011282 treatment Methods 0.000 description 14
- 208000009329 Graft vs Host Disease Diseases 0.000 description 9
- 208000024908 graft versus host disease Diseases 0.000 description 9
- 239000000427 antigen Substances 0.000 description 8
- 210000000612 antigen-presenting cell Anatomy 0.000 description 8
- 108091007433 antigens Proteins 0.000 description 8
- 102000036639 antigens Human genes 0.000 description 8
- 241000829111 Human polyomavirus 1 Species 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 241000701022 Cytomegalovirus Species 0.000 description 6
- 208000036142 Viral infection Diseases 0.000 description 5
- 230000035755 proliferation Effects 0.000 description 5
- 210000000130 stem cell Anatomy 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 230000001988 toxicity Effects 0.000 description 5
- 231100000419 toxicity Toxicity 0.000 description 5
- 241000701161 unidentified adenovirus Species 0.000 description 5
- 230000009385 viral infection Effects 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 4
- 239000002831 pharmacologic agent Substances 0.000 description 4
- VWFCHDSQECPREK-LURJTMIESA-N Cidofovir Chemical compound NC=1C=CN(C[C@@H](CO)OCP(O)(O)=O)C(=O)N=1 VWFCHDSQECPREK-LURJTMIESA-N 0.000 description 3
- 229960000724 cidofovir Drugs 0.000 description 3
- 239000012636 effector Substances 0.000 description 3
- 239000012467 final product Substances 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 210000000265 leukocyte Anatomy 0.000 description 3
- 210000003071 memory t lymphocyte Anatomy 0.000 description 3
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 2
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 2
- 241000701044 Human gammaherpesvirus 4 Species 0.000 description 2
- 102000043129 MHC class I family Human genes 0.000 description 2
- 108091054437 MHC class I family Proteins 0.000 description 2
- 108700018351 Major Histocompatibility Complex Proteins 0.000 description 2
- 230000006044 T cell activation Effects 0.000 description 2
- 230000000735 allogeneic effect Effects 0.000 description 2
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 210000003162 effector t lymphocyte Anatomy 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 210000005133 interdigitating dendritic cell Anatomy 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 210000005087 mononuclear cell Anatomy 0.000 description 2
- ZJAOAACCNHFJAH-UHFFFAOYSA-N phosphonoformic acid Chemical compound OC(=O)P(O)(O)=O ZJAOAACCNHFJAH-UHFFFAOYSA-N 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000020382 suppression by virus of host antigen processing and presentation of peptide antigen via MHC class I Effects 0.000 description 2
- 244000052613 viral pathogen Species 0.000 description 2
- 239000013603 viral vector Substances 0.000 description 2
- 206010065553 Bone marrow failure Diseases 0.000 description 1
- 206010014418 Electrolyte imbalance Diseases 0.000 description 1
- 206010029155 Nephropathy toxic Diseases 0.000 description 1
- IWUCXVSUMQZMFG-AFCXAGJDSA-N Ribavirin Chemical compound N1=C(C(=O)N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 IWUCXVSUMQZMFG-AFCXAGJDSA-N 0.000 description 1
- 230000006052 T cell proliferation Effects 0.000 description 1
- 230000005867 T cell response Effects 0.000 description 1
- WPVFJKSGQUFQAP-GKAPJAKFSA-N Valcyte Chemical compound N1C(N)=NC(=O)C2=C1N(COC(CO)COC(=O)[C@@H](N)C(C)C)C=N2 WPVFJKSGQUFQAP-GKAPJAKFSA-N 0.000 description 1
- 229960004150 aciclovir Drugs 0.000 description 1
- MKUXAQIIEYXACX-UHFFFAOYSA-N aciclovir Chemical compound N1C(N)=NC(=O)C2=C1N(COCCO)C=N2 MKUXAQIIEYXACX-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 208000035850 clinical syndrome Diseases 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 229960005102 foscarnet Drugs 0.000 description 1
- 229960002963 ganciclovir Drugs 0.000 description 1
- IRSCQMHQWWYFCW-UHFFFAOYSA-N ganciclovir Chemical compound O=C1NC(N)=NC2=C1N=CN2COC(CO)CO IRSCQMHQWWYFCW-UHFFFAOYSA-N 0.000 description 1
- 231100000304 hepatotoxicity Toxicity 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 210000001821 langerhans cell Anatomy 0.000 description 1
- 229960000681 leflunomide Drugs 0.000 description 1
- VHOGYURTWQBHIL-UHFFFAOYSA-N leflunomide Chemical compound O1N=CC(C(=O)NC=2C=CC(=CC=2)C(F)(F)F)=C1C VHOGYURTWQBHIL-UHFFFAOYSA-N 0.000 description 1
- 201000002364 leukopenia Diseases 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000011244 liquid electrolyte Substances 0.000 description 1
- 230000007056 liver toxicity Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000011278 mitosis Effects 0.000 description 1
- 231100000417 nephrotoxicity Toxicity 0.000 description 1
- 230000007694 nephrotoxicity Effects 0.000 description 1
- 208000004235 neutropenia Diseases 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229960000329 ribavirin Drugs 0.000 description 1
- HZCAHMRRMINHDJ-DBRKOABJSA-N ribavirin Natural products O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1N=CN=C1 HZCAHMRRMINHDJ-DBRKOABJSA-N 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 229960002149 valganciclovir Drugs 0.000 description 1
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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Cell Biology (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (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)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP19928301.1A EP4061386A4 (de) | 2019-11-19 | 2019-12-13 | Verfahren zur herstellung von multivirusspezifischen t-zellen |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| TR201918044 | 2019-11-19 | ||
| TR2019/18044 | 2019-11-19 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2021101466A1 true WO2021101466A1 (en) | 2021-05-27 |
Family
ID=75980745
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/TR2019/051071 WO2021101466A1 (en) | 2019-11-19 | 2019-12-13 | A method for producing multivirus specific t cells |
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 |
|---|---|---|---|---|
| ES2928851T3 (es) * | 2012-02-09 | 2022-11-23 | Baylor College Medicine | Mezclas pep para generar CTL multivíricos con amplia especificidad |
| AU2017326173B2 (en) * | 2016-09-16 | 2022-08-18 | Baylor College Of Medicine | Platform for activation and expansion of virus-specific T-cells |
-
2019
- 2019-12-13 EP EP19928301.1A patent/EP4061386A4/de active Pending
- 2019-12-13 WO PCT/TR2019/051071 patent/WO2021101466A1/en unknown
Non-Patent Citations (5)
| Title |
|---|
| FAIST BENJAMIN, SCHLOTT FABIAN, STEMBERGER CHRISTIAN, DENNEHY KEVIN M., KRACKHARDT ANGELA, VERBEEK MAREIKE, GRIGOLEIT GÖTZ U., SCH: "Targeted in-vitro-stimulation reveals highly proliferative multi-virus- specific human central memory T cells as candidates for prophylactic T cell therapy", PLOS ONE, vol. 14, no. 9, 30 September 2019 (2019-09-30), pages e0223258, XP055830387, DOI: 10.1371/joumal.pone.0223258 * |
| LITVINOVA L.S. ET AL.: "The influence of immunoregulatory cytokines IL -2, IL -7, and IL -15 upon activation, proliferation, and apoptosis of immune memory T-cells in vitro", CELL TISS. BIOL., vol. 7, 11 December 2013 (2013-12-11), pages 539 - 544, XP035366789, DOI: 10.1134/S1990519X13060072 * |
| QIAN C, WANG Y, REPPEL L, D'AVENI M, CAMPIDELLI A, DECOT V, BENSOUSSAN D: "Viral-specific T- cell transfer from HSCT donor for the treatment of viral infections or diseases after HSCT", BONE MARROW TRANSPLANT, vol. 53, no. 2, 23 October 2017 (2017-10-23), pages 114 - 122, XP055830388, DOI: 10.1038/bmt.2017.232 * |
| See also references of EP4061386A4 * |
| TESCHNER D. ET AL.: "Depletion of naive T cells using clinical grade magnetic CD 45RA beads: a new approach for GVHD prophylaxis", BONE MARROW TRANSPLANTATION, vol. 49, 12 August 2013 (2013-08-12), pages 138 - 144, XP055358248, DOI: 10.1038/bmt.2013.114 * |
Also Published As
| Publication number | Publication date |
|---|---|
| EP4061386A1 (de) | 2022-09-28 |
| EP4061386A4 (de) | 2024-01-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Szmania et al. | Isolation and expansion of cytomegalovirus-specific cytotoxic T lymphocytes to clinical scale from a single blood draw using dendritic cells and HLA-tetramers | |
| US20190024050A1 (en) | Enhanced generation of cytotoxic t-lymphocytes by il-21 mediated foxp3 suppression | |
| DE60026166T2 (de) | Vetozellen die wirksam in der prävention von transplantatabstossung sind und kein graft-versus-host potential aufweisen | |
| JP7423684B2 (ja) | 免疫細胞を操作するための抗原提示足場 | |
| JP2002515756A (ja) | 選択されたリンパ球の生産方法 | |
| JP5524056B2 (ja) | ＣＴＬとγδＴ細胞の同時誘導方法 | |
| US6821778B1 (en) | Methods for using dendritic cells to activate gamma/delta-T cell receptor-positive T cells | |
| Biondi et al. | Redirecting T cells with Chimeric Antigen Receptor (CAR) for the treatment of childhood acute lymphoblastic leukemia | |
| JP2018520095A5 (de) | ||
| EP2859092B1 (de) | Therapeutischer impfstoff zur behandlung von diabetes typ 1 bei kindern, anwendung eines zellsortierers und verfahren zur vermehrung von treg-zellen zur herstellung eines therapeutischer impfstoffs zur behandlung von diabetes typ 1 | |
| JP6942466B2 (ja) | 抗原特異的ｔ細胞受容体遺伝子を有する多能性幹細胞の製造方法 | |
| JP2017061558A (ja) | Ｔｈ１特性と細胞溶解性を発現する細胞 | |
| Yannelli et al. | The large scale generation of dendritic cells for the immunization of patients with non-small cell lung cancer (NSCLC) | |
| WO2005009466A1 (en) | Methods and compositions for increasing the efficiency of therapeutic antibodies using alloreactive natural killer cells | |
| EP4061386A1 (de) | Verfahren zur herstellung von multivirusspezifischen t-zellen | |
| JP2002501726A (ja) | 抗原提示および免疫応答を高める、改善された体外方法 | |
| US6056951A (en) | Method of reducing the chance of attack of cells in a body by the cellular immune system | |
| CA1333887C (en) | Immunity memory cell suspension and method of preparing same | |
| Introna et al. | Innovative cell-based therapies in onco-hematology: what are the clinical facts? | |
| CN102596209A (zh) | 含有短暂性存活ctl的医药品组合物 | |
| JP2639835B2 (ja) | 免疫記憶細胞懸濁液の調製方法 | |
| Sitkovskaya et al. | The Effect of Oncolytic Reovirus P-92 on Dendritic Cell Maturation and Generation of Tumor-Specific Lymphocytes in vitro | |
| WO2023238124A1 (en) | Virus-specific t cells, methods of their preparation and use thereof | |
| Valleca et al. | Mononuclear Cell Preparations | |
| Drawz et al. | Adoptive immunotherapy |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 19928301 Country of ref document: EP Kind code of ref document: A1 |
|
| NENP | Non-entry into the national phase |
Ref country code: DE |
|
| ENP | Entry into the national phase |
Ref document number: 2019928301 Country of ref document: EP Effective date: 20220620 |