# US 12,178,832 B2 · Methods for the production of TCR gamma delta + t cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,178,832 B2 - Document ID: US12178832B2 - Status: Granted - Kind: B2 - Application number: 17/307,754 - Filed: 2021-05-04 - Published: 2021-11-25 - Granted: 2024-12-31 - Earliest claimed: 2015-06-09 - Assignees: GammaDelta Therapeutics Ltd - Applicants: Gammadelta Therapeutics Ltd - Inventors: Diogo Antonio Remechido Anjos; Daniel Vargas Correia; Afonso Rocha Martins De Almeida - Web page: https://patentgrep.com/patent/US12178832B2 ## CPC - A61K 35/17: Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00) - A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells - A61K 40/31: Chimeric antigen receptors [CAR] - A61K 40/32: T-cell receptors [TCR] - A61K 40/42: Cancer antigens - A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics - A61P 35/00: Antineoplastic agents - A61P 35/02: specific for leukemia - A61P 37/04: Immunostimulants - C12N 5/0638: {Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]} - A61K 2039/5158: {Antigen-pulsed cells, e.g. T-cells} - C12N 2501/2301: Interleukin-1 (IL-1) - C12N 2501/2302: Interleukin-2 (IL-2) - C12N 2501/2304: Interleukin-4 (IL-4) - C12N 2501/2307: Interleukin-7 (IL-7) - C12N 2501/2315: Interleukin-15 (IL-15) - C12N 2501/2321: Interleukin-21 (IL-21) - C12N 2501/24: Interferons [IFN] - C12N 2501/515: CD3, T-cell receptor complex - C12N 2501/599: with CD designations not provided for elsewhere - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention relates to novel methods for the isolation and the selective ex vivo expansion of V32′ TCRy6+ T cells and to their clinical application. ## Claims ### Claim 1 A method of modulating an immune response comprising administering an effective amount of TCRγδ+ T cells obtained by a method for expanding Vδ2-TCRγδ+ T cells in a sample comprising: (1) culturing cells in the sample in a first culture medium comprising a T cell mitogen and interleukin-4; in the absence of interleukin-15, interleukin-2, or interleukin-7; and (2) culturing the cells obtained in step (1) in a second culture medium comprising a T cell mitogen and interleukin-15, interleukin-2, or interleukin-7, in the absence of interleukin-4, to an animal in need thereof. ### Claim 2 (depends on 1) A method according to claim 1, wherein the first or second culture medium, or both culture media, further comprise a second, a third and a fourth growth factor. ### Claim 3 (depends on 2) A method according to claim 2, wherein said growth factors are interferon-γ, interleukin-21 and interleukin-1β or a mimetic or functional equivalent thereof. ### Claim 4 (depends on 1) A method according to claim 1, wherein the first and second culture media further contain serum or plasma. ### Claim 5 (depends on 1) A method according to claim 1, wherein prior to step (1) the cells in the sample are enriched for T cells; enriched for TCRγδ+ T cells; depleted of TCRαβ+ T cells; first depleted of TCRαβ+ T cells, and then enriched for CD3+ cells; or depleted of non-TCRγδ+ T cells. ### Claim 6 (depends on 1) A method according to claim 1, wherein the sample is blood or tissue or fractions thereof. ### Claim 7 (depends on 6) A method according to claim 6, wherein the sample is selected from peripheral blood, umbilical cord blood, lymphoid tissue, epithelia, thymus, bone marrow, spleen, liver, cancerous tissue, infected tissue, lymph node tissue or fractions thereof. ### Claim 8 (depends on 1) A method according to claim 1, wherein in the first culture medium the T cell mitogen is present in an amount from about 10 to about 5000 ng/ml and interleukin-4 is present in an amount from about 1 to about 1000 ng/ml. ### Claim 9 (depends on 1) A method according to claim 1, wherein in the second culture medium the T cell mitogen is present in an amount from about 0.1 to about 50 μg/ml and interleukin-15 is present in an amount from about 1 to about 1000 ng/ml. ### Claim 10 (depends on 1) A method according to claim 1, wherein the T cell mitogen is an antibody or a fragment thereof. ### Claim 11 A method for treating an infection comprising administering an effective amount of TCRγδ+ T cells obtained by a method for expanding Vδ2-TCRγδ+ T cells in a sample comprising: (1) culturing cells in the sample in a first culture medium comprising a T cell mitogen and interleukin-4; in the absence of interleukin-15, interleukin-2, or interleukin-7; and (2) culturing the cells obtained in step (1) in a second culture medium comprising a T cell mitogen and interleukin-15, interleukin-2, or interleukin-7, in the absence of interleukin-4, to an animal in need thereof. ### Claim 12 A method for treating cancer comprising administering an effective amount of TCRγδ+ T cells obtained by a method for expanding Vδ2-TCRγδ+ T cells in a sample comprising: (1) culturing cells in the sample in a first culture medium comprising a T cell mitogen and interleukin-4; in the absence of interleukin-15, interleukin-2, or interleukin-7; and (2) culturing the cells obtained in step (1) in a second culture medium comprising a T cell mitogen and interleukin-15, interleukin-2, or interleukin-7, in the absence of interleukin-4, to an animal in need thereof. ### Claim 13 (depends on 12) The method according to claim 12, wherein the cancer is chronic lymphocytic leukemia, chronic myelogenous leukemia, acute myelogenous leukemia, acute lymphoblastic leukemia, and T cell and B cell leukemias, lymphomas (Hodgkin's and non-Hodgkin's), lymphoproliferative disorders, plasmacytomas, histiocytomas, melanomas, adenomas, sarcomas, carcinomas of solid tissues, hypoxic tumors, squamous cell carcinomas, genitourinary cancers such as cervical and bladder cancer, hematopoietic cancers, head and neck cancers, and nervous system cancers. ### Claim 14 A method for vaccinating an animal comprising administering an effective amount of TCRγδ+ T cells obtained by a method for expanding Vδ2-TCRγδ+ T cells in a sample comprising: (1) culturing cells in the sample in a first culture medium comprising a T cell mitogen and interleukin-4; in the absence of interleukin-15, interleukin-2, or interleukin-7; and (2) culturing the cells obtained in step (1) in a second culture medium comprising a T cell mitogen and interleukin-15, interleukin-2, or interleukin-7, in the absence of interleukin-4, to an animal in need thereof.