A kind of construction method of lrfft1 cell

Abstract

The present invention discloses a construction method of LRFFT1 cells. The construction method of the LRFFT1 cells comprises the following steps: ctDNA sequencing of human peripheral blood or whole exon sequencing of tumor tissues is conducted to screen out mutation sites for antigen epitope prediction, and a mutant polypeptide expression gene sequence is ligated and synthesized; simultaneously alentiviral vector is constructed, lentiviruses are packaged, the packaged lentiviruses transfect APC cells, modification of specific LV cells is completed, the APC cells are co-cultured with PBMC isolated from peripheral blood in vitro, effective peptides are screened out, and ordinary T cells are transformed into RFF cells with a more precise killing ability by a second impact of a precise and effective peptide stimulation; then a TCR-T technology principle is used to conduct modification; and the modified T cells are then subjected to a gene-editing technology to conduct knocking out of immunosuppressive targets, the T cells with the specific killing ability is precisely protected from in vivo inhibition, the killing ability of the T cells on tumor cells is improved, and the LRFFT1 cellsmore combining attack and defense are obtained.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for constructing LRFFT1 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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