Method for amplifying and multiplying T cells with antigenic specificity

A specific, cellular 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 it is difficult for immunogenic antigens to stimulate the specific proliferation of immune cells, etc. problem, to achieve the effect of good amplification effect, low cost and simple operation

Inactive Publication Date: 2012-08-08
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 lines expressing recombinant virus (containing the gene encoding anti-melanoma gp100 TCR protein)

[0035] APB plasmids were used to transfect Phoenix-Eco cells, and the resulting virus supernatants were used to infect PT67 packaging cells. The infected monoclonal PT67 virus supernatants were respectively infected with the SupT1 human lymphoma cell line to detect the expression of human CD3. Any infected SupT1 human lymphoma cell line served as blank control. This method is used to select the packaging cell line that produces the highest virus titer.

[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 pr...

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] The tetramer of the melanoma antigen gp100 peptide was prepared according to the literature (Estcourt, M.J., A.J.McMichael, and T.Hanke, Altered primary CD8+T cell response to a modified virusAnkara(MVA)-vectored vaccine in the absence of CD4+T cell help . Eur J Immunol, 2005.35 (12): p.3460-7) described in the method of preparation.

[0051] 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)-vectored vaccine in the absence off CD4+T Cell help. EurJ 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, the catalog number is 12-7311-71;

[0054] Monensin was purchased from eBioscience, the prod...

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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 invention relates to a method for amplifying and proliferating antigen-specific T cells. Background technique [0002] As an effective treatment, immunotherapy has been widely used in recent years. Among them, cellular immunotherapy is widely used in the treatment of tumors and cancers, and has achieved surprising results. But in the practice of cellular immunotherapy, there is a major obstacle, that is, it is difficult to produce sufficient numbers of immune cells capable of killing abnormal cells. At present, the expansion of immune cells is usually achieved through antibody stimulation and addition of cytokines. For example, the expansion of T cells is usually achieved by using anti-CD3 antibodies plus interleukin-2. 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 re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N15/09
CPCA61K2035/124C12N2501/515C12N5/0636C12N2510/00A61P31/04A61P31/12A61P37/04
Inventor 高斌丁洁
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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