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In vitro expansion method for anti-tumor T cells from non-sentinel lymph nodes

A technology of sentinel lymph nodes and lymphocytes, applied in the medical and health field, can solve the problems of reducing the anti-tumor ability of T cells, polarization of T cell subsets, weak anti-tumor function, etc. , the effect of maintaining anti-tumor activity

Active Publication Date: 2018-06-29
贵州太瑞生诺生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] (2) Inductive immunosuppressive effects (such as the increase of programmed death receptor 1, etc.) will be caused during the culture process, which will reduce the anti-tumor ability of T cells;
[0012] (3) The use of a single cytokine to expand T cells will cause the polarization of T cell subpopulations, making the anti-tumor function of a single subpopulation weaker than that of a mixed subpopulation

Method used

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Examples

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Effect test

experiment example

[0055] The number, proliferation efficiency, expression of immunosuppressive molecules and immune activation molecules and anti-tumor ability of non-sentinel lymph node-derived anti-tumor T cells amplified by Example 3 of the present invention were compared with those amplified by conventional methods, specifically: :

[0056] 1. About the number of cells:

[0057] Take the quantity as 1.6*10 6 Anti-tumor T cells derived from non-sentinel lymph nodes were expanded in vitro by the method described in Example 3 of the present invention, counted after the culture was completed, and the total amount of anti-tumor T cells derived from non-sentinel lymph nodes was 413*10 6 indivual.

[0058] Take the same quantity as 1.6*10 6 The non-sentinel lymph node-derived T cells were expanded in vitro by conventional methods, counted after the culture was completed, and the total amount of non-sentinel lymph node-derived anti-tumor T cells obtained was 45.6*10 6 indivual.

[0059] The re...

Embodiment 1

[0074] A method for in vitro expansion of anti-tumor T cells derived from non-sentinel lymph nodes, comprising the following steps:

[0075] (1) Antigen sensitization and immunosuppressive blockade:

[0076] a, get the lymphocyte culture medium that is 100:2 in volume ratio, fetal bovine serum mixes, obtain lymphocyte culture medium, fetal bovine serum mixture; Add interleukin 2 in lymphocyte culture medium, fetal bovine serum mixture, adjust all The concentration of said interleukin 2 in lymphocyte culture medium and fetal bovine serum mixture is 100 units / ml, to obtain culture medium A, for subsequent use;

[0077] b. Take part of the culture medium A, add non-sentinel lymph node-derived anti-tumor T cells, adjust the volume of the culture medium, so that the density of the non-sentinel lymph node-derived anti-tumor T cells in culture medium A is 1*10 6 cells / ml, cultivated for 12 hours;

[0078] c. Another part of the culture solution A was taken, and anti-programmed deat...

Embodiment 2

[0100] A method for in vitro expansion of anti-tumor T cells derived from non-sentinel lymph nodes, comprising the following steps:

[0101] (1) Antigen sensitization and immunosuppressive blockade:

[0102] a, get the lymphocyte culture medium that is 100:5 in volume ratio, fetal bovine serum mixes, obtain lymphocyte culture medium, fetal bovine serum mixture; Add interleukin 2 in lymphocyte culture medium, fetal bovine serum mixture, adjust all The concentration of interleukin 2 in lymphocyte culture medium and fetal calf serum mixture is 1000 units / milliliter to obtain culture medium A for subsequent use;

[0103] b. Take part of the culture medium A, add non-sentinel lymph node-derived anti-tumor T cells, adjust the volume of the culture medium, so that the density of the non-sentinel lymph node-derived anti-tumor T cells in culture medium A is 2*10 6 cells / ml, cultivated for 24 hours;

[0104] c. Another part of the culture solution A was taken, and anti-cytotoxic T lym...

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PUM

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Abstract

The invention discloses an in vitro expansion method for anti-tumor T cells from non-sentinel lymph nodes. The method comprises the following four steps: antigen-pulsed and immunosuppression blocking,T cell subset separation and excitated proliferation, reexcitation and rapid proliferation of a T cell subset, and harvesting. The in vitro expansion method has the characteristics that the activation of T cell specificity is facilitated, the anti-tumor capacity of T cells can be improved, the quantity of tumor specific T cells is increased, the operation is simple and convenient and the method is easy for popularization.

Description

technical field [0001] The invention belongs to the field of medical and health technologies, and in particular relates to an in vitro expansion method of non-sentinel lymph node-derived anti-tumor T cells. Background technique [0002] Tumor immunotherapy is the application of immunological principles and methods to improve the immunogenicity of tumor cells and / or sensitivity to lymphocyte killing, to stimulate and enhance the body's anti-tumor immune response, and to infuse immune cells and / or effector molecules In the host body, cooperate with the body's immune system to kill tumors and inhibit tumor growth. [0003] In many tumor immunotherapy programs, "adoptive T lymphocyte infusion" is an important one, which refers to the infusion of autologous or allogeneic anti-tumor T lymphocytes activated and expanded in vitro to patients to kill tumors in patients cellular process. [0004] In adoptive T cell immunotherapy, there are two main sources of T cells, namely periphe...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N2501/2302C12N2501/2307C12N2501/2321C12N2501/515C12N2501/998C12N2509/00
Inventor 杨远修瑾陈阶唐景玲
Owner 贵州太瑞生诺生物医药有限公司
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