Single-stranded TCR-T carrier and single-stranded TCR-T cell production process

Abstract

Provided are a single-stranded TCR-T carrier and a single-stranded TCR-T cell production process. According to the scheme, the process includes the steps of complete HPV specificity TCR selection, TCR-T carrier construction, TCR-T cell engineering and an activity test of artificially transformed TCR-T cells. A TCR sequence which can identify a major histocompatibility complex (MHC) as HLA-A2-presented HPV-16 E6 polypeptide is adopted and converted into functional single-stranded conformation, a novel stable assembly mode of TCR alpha links and TCR beta links is put forward in the carrier construction aspect, and a stable TCR-T retrovirus production cell system is used in the TCR-T cell engineering aspect. HPVTCR-T cells produced by means of the series of techniques can be effectively activated by recipient cells representing HPV virus polypeptide and can be applied to treatment of head and neck cancer, cervical cancer and other diseases which are caused by HPV.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the field of TCR-T immunotherapy technology and application. Background technique [0002] TCR-T technology is a cell therapy technology that artificially transforms autologous T cells to express tumor antigen-specific TCR. TCR-T cell therapy includes the selection of tumor-specific TCRs, the construction of TCR expression vectors, the reinfusion of cells after TCR-T transformation, and the monitoring of immune processes and other key technologies and therapeutic methods. Compared with the same type of CAR-T (chimeric antigen receptor T cell) cell therapy technology, TCR-T has a wider antigen selection space, so it can not only expand its applicable tumor range, but also reduce off-target effects [1]. However, although TCR-T has achieved some success in preclinical testing and treatment of a small group of patients [2,3], clinical trial data of TCR-T are still scarce. At...

AI Technical Summary

Problems solved by technology

The TCR-T clinical trials currently in progress use the expression vectors of the complete TCR α chain and β chain[2,3], but the vector sequence is long, and a special polypeptide-MHC is required to detect whether TCR is successfully introduced into T cells Tetramers will affect the expression and quality monitoring of TCR-T to a certain extent

Finally, the existing TCR-T technology directly introduces exogenous TCR into patient T cells, but does not effectively limit the mismatch between exogenous TCR and endogenous TCR

Mismatches may cause non-specific TCR recognition. Once a small number of cells recognize their own antigens and clone and proliferate, it will cause a large-scale autoimmune reaction, which will pose a great safety risk in clinical practice

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