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Method for non-therapeutic purpose restoration of immune function of infiltrating T lymphocytes

A lymphocyte and immune function technology, applied in the direction of animal cells, vertebrate cells, cell culture active agents, etc., can solve the problems that it is difficult to restore immune vitality and recovery

Inactive Publication Date: 2019-02-01
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Infiltrating T lymphocytes in primary liver cancer can only expand 20-fold in response to CD3 monoclonal antibody and other stimuli, and it is difficult to restore immune activity
Traditional methods are difficult to restore aging infiltrating T lymphocytes, such as blocking KLRG1, LAG3 and many other aging molecules with antibodies, it is difficult to restore

Method used

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  • Method for non-therapeutic purpose restoration of immune function of infiltrating T lymphocytes
  • Method for non-therapeutic purpose restoration of immune function of infiltrating T lymphocytes
  • Method for non-therapeutic purpose restoration of immune function of infiltrating T lymphocytes

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Experimental program
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Embodiment 1

[0049] 1. The peripheral blood of patients with primary liver cancer was collected in blood collection tubes anticoagulated with lithium heparin, and the peripheral blood mononuclear cells were rapidly separated by Ficoll-PaquePlus density gradient centrifugation. The operation steps were as follows:

[0050] 1) Dilute the blood with PBS equivalent to whole blood;

[0051] 2) Add the same amount of Ficoll separation solution as the diluted whole blood to the centrifuge tube, slowly add the diluted blood to the upper layer of the Ficoll separation solution along the tube wall, do not mix with the Ficoll separation solution, centrifuge at room temperature 1500rpm for 20min ;Centrifugal speed adopts slow rise and slow fall, the rising speed is set to 1, and the falling speed is set to 0;

[0052] 3) After centrifugation, it can be seen that the cells in the centrifuge tube are divided into four layers from top to bottom, the first layer is the plasma layer, the second layer is th...

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Abstract

Belonging to the technical field of T lymphocyte receptors, the invention provides a method for non-therapeutic purpose restoration of the immune function of infiltrating T lymphocytes. The method includes: mixing an immune function impaired infiltrating T lymphocyte solution with a blocking agent to obtain a blocking agent-containing solution, wherein the blocking agent includes an anti-Tim-3 blocking antibody and / or an anti-PD-1 blocking antibody; mixing the blocking agent-containing solution with an anti-CD3 antibody and an anti-CD28 antibody, and then performing cultivation to obtain a culture; and mixing the culture with Brefeldin A, and performing incubation to obtain immune function restored infiltrating T lymphocytes. The method provided by the invention can restore the immune function of immune function impaired infiltrating T lymphocytes.

Description

technical field [0001] The invention belongs to the technical field of T lymphocyte receptors, and in particular relates to a method for restoring the immune function of infiltrating T lymphocytes for non-therapeutic purposes. Background technique [0002] At present, the etiology of most primary liver cancer patients in my country is hepatitis B virus infection. The prior art records that infiltrating T lymphocytes (TILs) in primary liver cancer (HCC) tissue express a large number of senescent molecules. In a state of immune incompetence and aging. Infiltrating T lymphocytes can induce tolerance in chronic hepatitis virus infection at an early stage. After primary liver cancer, infiltrating T lymphocytes are further induced in the tumor microenvironment to be incapacitated and senescent. Resistant to the organ environment, infiltrating T lymphocytes in primary liver cancer are more difficult to activate and expand than infiltrating cells in other tumors. The infiltrating T...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N2501/51C12N2501/515
Inventor 胡安斌刘芙蓉凌象超
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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