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A method for preparing antigen-specific T cells with NK cell characteristics

A kind of NK cell and specific technology, which is applied in the field of in vitro expansion of immune cells, can solve the problems of insufficient recognition range and slow proliferation, and achieve the effect of increasing cell proliferation speed, low expression level, and increasing proliferation speed

Active Publication Date: 2019-10-29
上海闪锦生物科技有限公司
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Problems solved by technology

[0005] In order to overcome the disadvantages of antigen-specific T cells cultured by conventional methods, such as insufficient recognition of cancer cells, slow proliferation, and large expression of PD-1 molecules, the present invention provides a high-efficiency preparation method of antigen-specific T cells. The resulting T cells express a large amount of CD56, which not only maintains the lethality against cancer cells with antigen-presenting ability, but also has non-MHC-restricted lethality

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  • A method for preparing antigen-specific T cells with NK cell characteristics
  • A method for preparing antigen-specific T cells with NK cell characteristics
  • A method for preparing antigen-specific T cells with NK cell characteristics

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Embodiment Construction

[0017] The present invention is specifically described below with examples, but the present invention is not limited thereto. In the following specific steps, the experimental methods that do not specify specific conditions shall be carried out according to the conventional conditions or the conditions suggested by the manufacturer.

[0018] 1. Use the present invention to prepare Epstein-Barr virus-specific T cells with NK cell characteristics.

[0019] (1) 100 ml of peripheral blood was drawn from volunteers, and mononuclear cells (PBMC) were separated by density gradient centrifugation with Ficoll-PlaquePlus, a lymphocyte separation medium, washed twice with PBS, and the resulting cells were cryopreserved.

[0020] (2) Take 1×10 6 The peripheral blood mononuclear cells were infected with Epstein-Barr virus and cultured in a 37°C, 5% CO2 incubator for 4 weeks to obtain a B lymphoblastoid cell line (LCL) transformed from B cells. LCL can present Epstein-Barr virus-related a...

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Abstract

The invention discloses a preparation method of antigen specific T cells with NK cell traits. T cells and antigen-presenting cells for presenting specific antigens are co-cultured according to a certain ratio, and a specific dosage of IL-15 is added so as to activate and quickly amplify the specific T cells. The obtained T cells retain the specific killability for tumor cells which present the antigens on the surfaces of the cells, also have the NK cell traits, can express CD56, are extremely high in non-MHC restrictive killability, and can effectively kill the tumor cells which do not express MHC.

Description

technical field [0001] The invention belongs to the field of immunotherapy, in particular to a culture method for immune cell expansion in vitro. Background technique [0002] As a new type of anti-cancer therapy, immunotherapy, together with traditional surgery, chemotherapy, and radiotherapy, constitutes the four pillars of cancer treatment. In immunotherapy, cellular immunotherapy shows great development potential. According to the immune cells used, cellular immunotherapy can be divided into two types. One type uses non-specific immune cells, such as NK cells and CIK (CD3+CD56+) cells, which are non-MHC-restricted in killing cancer cells, and the recognized surface targets of cancer cells are not antigens presented by MHC Fragments, but targets recognized by NK cells such as NKG2D, DNAM-1, and NKp30 (Pievani A, Borleri G, et al. Blood. 2011, 118(12):3301-3310). Another type of therapy uses specific immune cells, such as chimeric antigen receptor-modified T cells, TCR-...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2501/2315C12N2502/1107C12N2502/1114C12N2502/1121
Inventor 罗益华吴春晓赵剑锋
Owner 上海闪锦生物科技有限公司
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