Preparation method of dendritic cells and application thereof

Abstract

The invention provides a preparation method of dendritic cells, which belongs to the technical field of tumor treatment. The preparation method comprises the steps of: 1) ligating a fusion gene of human leukocyte antigen (HLA) and KRAS mutation to pCDH or a mammalian expression vector to obtain a recombinant plasmid; 2) transforming the dendritic cells (DC) derived from peripheral blood by electroporation with the recombinant plasmid to obtain the recombinant cells (i-DC); and 3) inducing maturation of the recombinant cells to obtain the mature dendritic cells. In the prior art, antigen-presenting cells are generally loaded by mutating polypeptides, or mutant antigens are transferred into antigen-presenting cells, but the efficiency of both is low, The method converts the fusion gene of KRAS antigen and HLA into the antigen-presenting cells and overexpresses two genes, increases the binding rate of HLA and KRAS mutant antigen, improves the efficiency of presentation, and more easily obtains CTL with specific killing effect.

Description

technical field [0001] The invention relates to the technical field of tumor treatment, in particular to a preparation method and application of dendritic cells. Background technique [0002] There are three kinds of genes related to human tumors in the ras gene family: HRAS, KRAS mutation and NRAS, which are located on chromosome 11, 12 and 1 respectively. The KRAS mutation is also known as the p21 gene because it encodes a 21kD ras protein. Among the ras genes, KRAS mutations have the greatest impact on human cancer. When KRAS mutations are normal, they can control the pathways that regulate cell growth; when abnormal, they cause cells to continue to grow and prevent cells from self-destruction. It participates in intracellular signal transmission. When the KRAS mutation gene is mutated, the gene will be permanently activated, unable to produce normal ras protein, causing intracellular signal transduction disorder, cell proliferation out of control and cancerous. [0003...

AI Technical Summary

Problems solved by technology

Therefore, patients with KRAS mutations are difficult to benefit from targeted drugs and have a short survival period. Therefore, it is urgent to develop immune cell therapy targeting KRAS mutations; the traditional preparation of cytotoxic T lymphocytes (CTL) targeting KRAS mutations On the one hand, it is affected by the weak presentation ability of the patient's dendritic cells (DC), and on the other hand, it is affected by the lack of the patient's human leukocyte antigen (HLA), resulting in the preparation of CTLs that have no ability to kill tumor cells with KRAS mutations.

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