LRFF cell construction method

Abstract

The invention provides an LRFF cell construction method, human peripheral blood is used to sequence ctDNA or a whole exome of a tumor tissue, a mutation site is screened to predicate an epitope, and the sequence of an expression gene of a mutation polypeptide is connected and synthesized; meanwhile, a lentiviral vector is constructed, a lentivirus is packaged, an APC (Antigen Presenting Cell) is transfected to finish the transformation of a specific LV cell, the APC is cultured in vitro together with a PBMC (Peripheral Blood Mononuclear Cell) separated from the peripheral blood, the effectiveaccurate polypeptide is screened, a common T cell is transformed into an LRFF cell having more accurate lethality through the second impact stimulated by the accurate polypeptide; and the lethality ofthe T cell on a tumor cell is improved. The provided LRFF cell is widely applied to the individualized accurate treatment of a solid tumor.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for constructing LRFF 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. [0003] The above treatment methods have limitations, especially the in vitro induction of DC cells and the technology of DC cell loading tumor antigens have been studied theoretically, but there are still m...

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Problems solved by technology

[0003] The above treatment methods have limitations, especially the in vitro induction of DC cells and the technology of DC cell loading tumor antigens have been studied theoretically, but there are still many problems in the actual implementation process, lack of clear signal transduction that is critical for the development of tumor cells Pathway-related molecules are used as inducing antigens. Due to unknown tumor antigens and obstacles to immunosuppression in the tumor microenvironment, it is difficult to implement specific cell-targeted immunotherapy

In addition, although some antigens have been impacted in vitro, they have not undergone in vitro co-cultivation and in vitro expansion, allowing relatively thin specific cells to directly face the complex tumor immune microenvironment. Therefore, it is difficult to achieve the expected effect

Although some can also be presented and co-cultivated in vitro, they have a single target (MAGE-3) and are only effective for certain cancer types such as non-small cell lung cancer.

Although there are also attempts to transfect and present lentivirus as a vector, the safety and convenience are not as good as the polypeptide method

The direct stimulation of simple mixed peptides is simple and convenient, but the efficiency is low

[0004] The above-mentioned schemes generally lack accurate and effective analysis of patient antigens. A more effective way should not only consider safety issues, but also take into account the efficiency of antigen presentation, so as to achieve the goal of precise treatment

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