Method for amplification of gamma delta T cells by phosphate and application thereof

Abstract

The invention provides a method for in vitro amplification of human gamma delta T cells and application thereof. The method for in vitro amplification of human gamma delta T cells includes the steps of: (1) preparing or providing mononuclear cell; (2) adding the mononuclear cell into a cell culture liquid, and placing the cell culture liquid into a cell incubator to conduct culture; and (3) acquiring the cells. The cell culture liquid contains (1-hydroxy-3-(methyl pentyl amino)propylidene)diphosphonic acid monosodium salt (IBA) and human recombinant interleukin rhIL-2. According to the invention, the method of combining rhIL-2 and IBA to stimulate human peripheral blood mononuclear cells (PBMC) is employed to amplify human gamma delta T cells in vitro. The method has the characteristics of low cost, simple preparation and high amplification efficiency, and can obtain gamma delta T cells with strong tumor lethality and TCR (T cell receptor) cloning diversity.

Description

technical field [0001] The invention relates to the field of immunology, in particular to a method for amplifying γδT cells with phosphate and application thereof. Background technique [0002] T lymphocytes are divided into two types according to the expression of T cell antigen receptor (TCR): TCRαβT cells and TCRγδT cells, referred to as αβT cells and γδT cells, respectively. γδT cells have the ability to specifically recognize antigens without MHC restriction, have anti-infection, anti-tumor and immune regulation functions, and are the first line of defense for the body's immune surveillance. How to efficiently and rapidly expand human peripheral blood γδT cells in large quantities in vitro is very important for the study of γδT cell immune function, improving patient immunity, anti-tumor and anti-infection capabilities, but due to the high content of γδT cells in peripheral blood and tumor tissues It is not high, generally no more than 5% of the total number of T cells...

AI Technical Summary

Problems solved by technology

How to efficiently and rapidly expand human peripheral blood γδT cells in large quantities in vitro is very important for the study of γδT cell immune function, improving patient immunity, anti-tumor and anti-infection capabilities, but due to the high content of γδT cells in peripheral blood and tumor tissues Not high, generally no more than 5% of the total number of T cells, it is very difficult to obtain a large number of γδT cells with high cytotoxic activity

[0003] The invention patent with the authorized announcement number CN1306027C provides a method for expanding γδT lymphocytes in vitro. This method uses solid-phase anti-γδTCR antibody to selectively expand γδT cells. However, the anti-γδTCR antibody used is expensive and the production cost is too high.

[0004] The invention patent with the authorized announcement number CN102533649B provides a simple and efficient method for preparing human γδT cells. The method uses Mycobacterium tuberculosis thermostable antigen combined with cytokines human recombinant rhIL-2 and human recombinant rhIL-21 to stimulate and expand γδT cells. However, the specific clones of γδT cells obtained by this method are dominant, but the universality is poor, and it cannot widely kill different types of tumors

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