Phenotypic and functional features of in vitro generated TCR-T cells specific for the tumor-associated antigen NY-ESO-1

Abstract

Introduction. The antitumor activity of T cells holds great potential for the development of immunotherapeutic schemes for the treatment of oncological diseases. Gene modification has become a new milestone in studies of antitumor T-cell immunity, which makes it possible to obtain a population of cells with a T cell receptor (TCR) of a given specificity from a heterogeneous pool of T lymphocytes. In this work, using a gammaretrovirus vector encoding a TCR specific for the epitope of the tumor-associated NY-ESO-1 antigen, NY-ESO-1 specific TCR-T cells were obtained, the features of their phenotype were studied in vitro, and the antigen-dependent activation and antitumor cytotoxic activity.

Aim of the study – to research the phenotypic and functional features of in vitro generated TCR-T lymphocytes for the tumor-associated antigen NY-ESO-1.

Material and methods. The phenotype of transduced cells was studied using flow cytometry by expression of markers of memory cells (CD45RA and CD62L), activation and depletion (CD69, CD95, PD-1, TIM-3). The cytotoxic potential of the obtained cells was assessed using the analysis of the release of lactate dehydrogenase in conditioned media from cultivation with target cells of tumor lines SK-Mel-37 and NW-Mel-38. Additionally, after cultivation with target cells, the expression of markers of cell activation and cytotoxicity (4-1BB, CD69, CD40L, FasL) was assessed using flow cytometry.

Results. Transduced NY-ESO-1-specific TCR-T cells were predominantly poorly differentiated T-lymphocytes with high viability and were characterized by a phenotype shift towards central and effector memory cells compared to non-transduced T-lymphocytes. Shown antigen-specific cytotoxic response to cultivation with tumor cells positive for the expression of NY-ESO-1. When cultured with the SK-Mel-37 line, a significant increase in the proportion of cells expressing CD40L and FasL was observed compared to the proportion in the absence of target cells, which confirms the activation of the effector properties of the transduced T cells in response to the antigen.

Conclusion. In vitro generated NY-ESO-1-specific TCR-T cells are characterized by a poorly differentiated memory phenotype and a high level of viability, and have a cytotoxic potential against tumor lines expressing NY-ESO-1.

Keywords:TCR; TCR-T cells; NY-ESO-1; memory T cell subsets; T cell activation; cellular cytotoxicity

For citation: Kuznetsova M.S., Tereshchenko V.P., Shevchenko J.A., Fisher M.S., Kurilin V.V., Alsalloum A., Alrhmoun S., Akahori Y., Shiku H., Sennikov S.V. Phenotypic and functional features of in vitro generated TCR-T lymphocytes specific for the tumor-associated antigen NY-ESO-1. Immunologiya. 2022; 43 (5): 536–47. DOI: https://doi.org/10.33029/0206-4952-2022-43-5-536-547 (in Russian)

Funding. The study was supported by the grant of the Russian Science Foundation (project No. 21-65-00004). URL: https://rscf.ru/project/21-65-00004/

Conflict of interests. Authors declare no conflict of interests.

Authors’ contributions. The concept and design of the study – Kuznetsova M.S., Shevchenko J.A., Tereshchenko V.P., Kurilin V.V., Akahori Y., Shiku H., Sennikov S.V.; data collection and processing – Kuznetsova M.S., Shevchenko J.A., Tereshchenko V.P., Fisher M.S., Alsalloum A., Alrkhmoun S.; statistical data processing – Kuznetsova M.S., Shevchenko J.A.; text writing – Kuznetsova M.S.; editing Kuznetsova M.S. Sennikov S.V.; approval of the final version of the article – Sennikov S.V.; responsibility for the integrity of all parts of the article – Kuznetsova M.S.

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