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Method for amplifying and multiplying T cells with antigenic specificity

A specific and cell-based technology, applied in animal cells, antibacterial drugs, vertebrate cells, etc., can solve the problem that the number of effective target cells is small, the treatment cannot be satisfied, and immunogenic antigens are difficult to stimulate the specific proliferation of immune cells, etc. problem, to achieve good amplification effect, low cost, and simple operation

Inactive Publication Date: 2010-09-08
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method can only expand immune cells non-specifically, and the number of effective target cells obtained is not large, which cannot meet the needs
Another method is to perform selective amplification, that is, to repeatedly stimulate immune cells with antigen-presenting cells (such as dendritic cells, B cells, etc.) that bind to the target antigen to obtain a large number of target antigen-specific immune cells, but this The method is more suitable for the expansion of specific immune cells with strong immunogenic antigens, and it is difficult for weak immunogenic antigens to stimulate immune cells to proliferate in large quantities, which cannot meet the needs of treatment

Method used

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  • Method for amplifying and multiplying T cells with antigenic specificity
  • Method for amplifying and multiplying T cells with antigenic specificity
  • Method for amplifying and multiplying T cells with antigenic specificity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Preparation of bispecific T cells (anti-tumor-specific and anti-influenza virus-specific T cells)

[0034] 1. Preparation of packaging cell line expressing recombinant virus (containing gene encoding anti-melanoma gp100TCR protein)

[0035] Phoenix-Eco cells were transfected with APB plasmid, and the resulting virus supernatant was used to infect PT67 packaging cells. The monoclonal PT67 virus supernatants after infection were respectively infected with SupT1 human lymphoma cell line, and the expression of human CD3 was detected. Any infected SupT1 human lymphoma cell line is a blank control. This method is used to screen for packaging cell lines that produce the highest viral titers.

[0036] The result is as figure 1 As shown in A, it shows that the screened PT67 packaging cell line produces the highest virus titer (that is, it can maximize the expression of human CD3 on the surface of SupT1 cells), and can be used as a packaging cell line for toxin produ...

Embodiment 2

[0048] Example 2. Proliferation and function of bispecific T cells

[0049] The bispecific T cells obtained in Example 1 were subjected to the following experiments.

[0050] Tetramers of melanoma antigen gp100 peptides were as described in the literature (Estcourt, M.J., A.J. McMichael, and T. Hanke, Altered primary CD8+ T cell response to a modified virus Ankara (MVA)-vectored vaccine in the absence of CD4+ T cell help. Prepared as described in Eur J Immunol, 2005. 35(12): p.3460-7).

[0051] Tetramers of the influenza virus antigen matrix protein MP peptide were prepared according to the literature (Estcourt, M.J., A.J. McMichael, and T. Hanke, Altered primary CD8+ T cell response to a modified virus Ankara (MVA)-vectoted vaccine in the absence of CD4+ T cell Prepared as described in help. Eur J Immunol, 2005. 35(12): p. 3460-7).

[0052] FITC-labeled anti-mouse CD8 was purchased from BD, PharMingen, CA, USA;

[0053] Anti-mouse IFN-γ-PE was purchased from eBioscience, c...

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Abstract

The invention discloses a method for amplifying and multiplying T cells with antigenic specificity, which comprises the following steps of: stimulating the T cells by using an immunogen A to obtain the T cells with the immunogen A specificity aiming at a polypeptide-MHC molecular compound of the immunogen A; transferring a coding gene of an immunological identification molecule for recognizing a polypeptide-MHC molecular compound of a target antigen B into the T cells with the immunogen A specificity to obtain the T cells with bi-anti-specificity for recognizing the polypeptide-MHC molecular compound of the target antigen B and the polypeptide-MHC molecular compound of the immunogen A; and stimulating to amplify and multiply the T cells with bi-anti-specificity by using the immunogen A. The T cells with bi-anti-specificity prepared by the method simultaneously have the target antigen B specificity and the immunogen A specificity; the induction and expanding culture are performed by using the immunogen A so as to obtain a great number of immune cells with the target antigen B specificity; the defect that the conventional antigen with weak immunogenicity cannot induce the specific immune cells to multiply in vitro is overcome; and the multiplied T cells with the bi-anti-specificity have high kill rate for the target antigen.

Description

technical field [0001] The present invention relates to a method for amplifying and proliferating antigen-specific T cells. Background technique [0002] As an effective treatment method, immunotherapy has been widely used in recent years. Among them, cellular immunotherapy is widely used in the treatment of tumor and cancer, and has achieved surprising results. However, in the practice of cellular immunotherapy, there is a major obstacle, that is, it is difficult to produce sufficient numbers of immune cells that can kill abnormal cells. At present, the expansion of immune cells is usually achieved by antibody stimulation and the addition of cytokines. For example, the expansion of T cells is usually achieved by anti-CD3 antibody and interleukin-2. However, this method can only amplify immune cells non-specifically, and the number of effective target cells obtained is small, which cannot meet the needs. Another method is to carry out selective expansion, that is, to repe...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/09
CPCA61K2035/124C12N2501/515C12N5/0636C12N2510/00A61P31/04A61P31/12A61P37/04A61K2239/57A61K39/4611A61K2239/31A61K39/464838A61K2239/38A61K39/4632A61K39/464492
Inventor 高斌丁洁
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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