Tumor neoantigen specific tumor infiltration lymphocyte co-culture method

Abstract

The invention relates to the technical field of cell culture, in particular to a tumor neoantigen specific tumor infiltration lymphocyte co-culture method which is characterized by comprising the following steps: S1, tumor neoantigen preparation: respectively collecting the tissues of lung cancer (N1), pancreatic cancer (N2), bile duct cancer (N3), triple-negative breast cancer (N4) and melanoma tumor (N5), separating tumor cells, andcollecting normal tissues of corresponding individuals; comparing the all-exon sequencing and normal tissue and tumor tissue sequencing results to determine all amino acid mutations, then designing a gene construct containing all mutant peptide sequences and synthesizing a long-chain polypeptide according to a neoantigen coded by a mutant gene. The TIL cells harvested by designing a co-culture method of tumor neoantigen specific tumor infiltration lymphocytes can effectively kill autologous tumor target cells, can induce immune cells to have multiple target spots, are free of limitation of cancer species, and have an immune memory effect.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for co-cultivating tumor neoantigen-specific tumor infiltrating lymphocytes. Background technique [0002] Tumor infiltrating lymphocyte immunotherapy, as early as 1986, was first developed by Steve Rosenberg of the National Cancer Institute (NCI) and used to treat some solid tumors (about 90% of cancers in the world are solid tumors). As of 2019, clinical research results show that 66 The ORR (Objective Response Rate) of unresectable melanoma patients was 38% (2 CRs, 23 PRs), and the DCR (Disease Control Rate) was 80%. TIL therapy is effective in melanoma, cervical cancer, head and neck cancer, lung cancer, The clinical curative effect of breast cancer, cholangiocarcinoma, and colorectal cancer is impressive. Typical cases: 1 case of advanced breast cancer with 42-month tumor-free survival, 1 case of advanced cholangiocarcinoma who failed chemotherapy and survived ...

AI Technical Summary

Problems solved by technology

[0006] Tumor neoantigen (neoantigen) is ubiquitous in tumor cells: During the process of tumorigenesis, cancer cells will produce many gene mutations, some of which are not found in normal tissues and cells. These proteins will activate the immune system and cause the immune system to attack cancer cells. In the past, most cancer vaccines mostly used tumor-associated antigens Taa (Tumoraassociated antigens), which were expressed in normal tissues but highly expressed in cancer tissues, so the immune system regarded these self-antigens as ordinary foreign bodies and did not produce a strong immune response Under certain conditions, it may cause abnormalities in the autoimmune system; therefore, the curative effect is not obvious. Tumor neoantigens are somatic mutation products that do not exist in normal tissues and cells. They have high immunogenicity and specificity. The preparation process of individual neoantigen vaccines has been completed. Gradually systematize the identification of neoantigens, first use whole exome sequencing to compare the gene sequences of cancer and normal tissues, obtain the data of mutation genes in cancer tissues, select the appropriate mutant neoantigens according to the gene expression level, and use bioinformatics or artificial intelligence algorithms , to predict the adhesion affinity between the candidate and the human leukocyte antigen (HLA), based on which the candidate antigen sequence is further selected and those neoantigens are finally selected, and the long-chain polypeptide vaccine is synthesized according to the neoantigen encoded by the mutated gene, and the immune adjuvant is added to further enhance the immunity Originality, can induce multi-target immune cells, no cancer type restriction, and has immune memory effect. At present, there is a lack of a culture method for TIL cells, which can specifically and effectively kill autologous tumor target cells, and can induce multi-target immune cells. point, no cancer type restriction, immune memory function