Construction method of LRFFT2 cells

Abstract

The invention provides a construction method of LRFFT2 cells; human peripheral blood is used for ctDNA sequencing or tumor tissues are used for full exon sequencing, mutant sites are screened out forepitope prediction and are connected to synthesize a mutant polypeptide expression gene sequence; at the same time, a lentivirus vector is constructed, lentiviruses are packaged, APC cells are transfected, the transformation of specific LV cells is completed, the transformed specific LV cells are co-cultured with PBMC isolated from the peripheral blood in vitro, and effective polypeptides are screened out; through precise effective second shocking stimulated by the polypeptides, ordinary T cells are transformed into RFF cells with more precise killing ability; transformation is preformed by aTCR-T technical principle; the transformed T cells are subjected to immunosuppressive signal sealing with antibody drugs in vitro, the specific killer T cells are precisely protected from inhibition in vivo, the killing ability of the T cells to tumor cells is improved, and the LRFFT2 cells with both attack and defense are obtained. The LRFFT2 cells can be widely applied in individualized and precise treatment of solid tumors.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for constructing LRFFT2 cells. Background technique [0002] At present, in terms of tumor-specific immunotherapy, the existing LAK, DC, CIK, and DC-CIK cells and methods have basically been proved to be ineffective, while NK, CAR-NK, TIL and other cell technologies have yet to mature. CAR- T cells also have shortcomings in safety and solid tumor therapy. In the prior art, DCs are generally modified to present T cells to produce specific killing. Some laboratories are trying to use viruses as vectors to transfect and present T cells to induce specific killing of T cells. We have also used mutant hybrid peptides to directly stimulate PBMCs to induce T cells. There are also laboratories using TCR-T technology to target the presentation of MAGE A3 antigen. [0003] The above treatment methods are immature, especially the in vitro induction of DC cells and the te...

AI Technical Summary

Problems solved by technology

There is a general lack of accurate and effective analysis of patient antigens

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