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Gene modified CDR3 delta transplanted gamma delta T lymphocyte and its antitumor purpose

A technology of gene modification and lymphocytes, applied to cells modified by introducing foreign genetic material, biochemical equipment and methods, microorganisms, etc. drop and other issues

Active Publication Date: 2014-08-13
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there are still some problems to be solved urgently in the preparation of γδT cell preparations for treatment
For example, in patients with advanced tumors or after chemotherapy, immunosuppression in the body leads to a decrease in the number of seed cells and a decline in immune activity, resulting in a limited number of expanded cells in vitro, and it is difficult to achieve the amount of cell preparation required for treatment.

Method used

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  • Gene modified CDR3 delta transplanted gamma delta T lymphocyte and its antitumor purpose
  • Gene modified CDR3 delta transplanted gamma delta T lymphocyte and its antitumor purpose
  • Gene modified CDR3 delta transplanted gamma delta T lymphocyte and its antitumor purpose

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Embodiment Construction

[0029] 1. Materials and methods

[0030] 1. Cell culture

[0031] PBMC, freshly isolated normal human peripheral blood mononuclear cells; RetroPack TM PT67 is a packaging replicating retroviral cell line derived from NIH3T3, purchased from Clontech Laboratories, Inc. NIH3T3 is a mouse fibroblast cell line. SKOV 3 , human ovarian serous papillary cystadenocarcinoma cell line; HO8910, human ovarian serous cystadenocarcinoma cell line; Hela, human cervical cancer tumor cell line; Daudi, human B lymphoma cell line; Raji, human Burkitt lymphoma cell line Both can be purchased from the Cell Center, School of Basic Science, Chinese Academy of Medical Sciences. SKOV3 cells were cultured adherently in McCoy 5A medium containing 10% FCS. Hela, NIH3T3 and RetroPack PT67 cells were cultured adherently in DMEM high glucose medium containing 10% FCS. HO8910, Raji, and Daudi, in RPMI-1640 medium containing 10% FCS for adherent or suspension culture.

[0032] 2. Animals

[0033] BALB...

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Abstract

The invention relates to a Gene modified CDR3 delta transplanted gamma delta T lymphocyte and its antitumor purpose. Retrovirus particles of a gamma delta TCR delta 2 chain and a gamma delta TCR delta 9 chain which contains an OT 3 sequence (FPSHTFHSTVVHT) are respectively packaged, the retrovirus particles are used for infecting normal human PBMC stimulated by an anti-CD3 antibody, the cell membrane surface expresses TCR gamma delta, the gene modified CDR3 delta transplanted gamma 9 delta 2T lymphocyte can be obtained, experiment proves that the cells possess cytotoxic effect on a plurality of tumor cells. The gene modified CDR3 delta transplanted gamma delta T lymphocyte and its antitumor purpose provide a novel method and a strategy for adoptive immunotherapy of the tumor.

Description

technical field [0001] The present invention relates to the fields of genetically modified cells and tumor treatment. Specifically, the present invention relates to the field of genetically modifying T lymphocytes and treating tumors with the modified γ9δ2 T lymphocytes. Background technique [0002] Immunotherapy of tumors has high hopes, especially the adoptive immunotherapy of tumor-specific T lymphocytes is considered to be a potential treatment strategy. However, the current experimental or clinical trials have not shown satisfactory efficacy. This may be closely related to the extremely weak or even absent antigenicity of most tumors, the defect of antigen presentation function, the low expression level of surface MHC molecules or co-stimulatory molecules, and the immunosuppressive state of tumor-bearing organisms. In addition, the number of tumor-specific cytotoxic T lymphocytes (CTL) in cancer patients is small, and it is difficult to isolate and expand in vitro; i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N7/01A61K35/14A61P35/00A61P35/02A61P15/00A61K35/15
Inventor 何维赵慧郗雪艳崔莲仙
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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