Acquisition method and application of individualized tumor neoantigen specific CD8 cells

Abstract

The invention relates to an acquisition method and application of individualized tumor neoantigen specific CD8 cells. The acquisition method comprises the following steps: 1) injecting tumor neoantigen vaccines into a patient, and then sampling blood; 2) obtaining peripheral blood mononuclear cells by a density gradient centrifugation method, and cryopreserving the obtained peripheral blood mononuclear cells; (3) performing mononuclear cell culture; (4) adding monophosphoryl lipid A and gamma interferon for culture; (5) collecting mature DC cells for continuous culture; (6) resuscitating the cryopreserved resuscitated lymphocytes, mixing the resuscitated lymphocytes with the DC cells in proportion, continuing to culture, and adding human recombinant interleukin 2 on the third day and the tenth day; and (7) on the seventh day of co-culture of the DC cells and the lymphocytes, resuscitating the cryopreserved antigen-loaded DC cells, performing adding for co-culture on the fourteenth day, and harvesting the antigen-specific CD8 CELLs. According to the method, the specific antigen is obtained through individualization, and the CD8 cells are activated and tumor cells are killed through the antigen presented by the DC cells, so that tumor growth is more effectively controlled.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for obtaining individualized tumor neoantigen-specific CD8 cells. Background technique [0002] In vitro generation and expansion of T cells (CTL, cytotoxic T lymphocyte, cytotoxic lymphocyte) with the ability to recognize tumor antigens have been successfully used to treat malignant tumors and viral infectious diseases. Tumor neoantigens, that is, proteins / peptides encoded by tumor mutation genes, have recently become the main tumor-specific antigens used in this field. The attractiveness of this specific adoptive immune cell therapy strategy is that it can induce a specific immune response only against diseased cells, without harming normal cells and without side effects. Therefore, a routine and reproducible method for inducing and expanding tumor neoantigen-specific CTL in vitro is the key to realize the adoptive therapy of tumor immune cells. [0003] At present, the ...

AI Technical Summary

Problems solved by technology

However, due to the relatively short survival period of DC cells, in the process of cultivating CTLs, it is necessary to continuously collect and replenish dead DC cells, activate and expand them multiple times to obtain clinical treatment, which requires a lot of manpower, which is time-consuming and costly. Poor repeatability, high chance of contamination, uncertain curative effect, etc.

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