Modelling antigen-specific T cell-mediated killing of cancer cells in vitro.

Antigen-specific T cell responses are critical for effective cancer cell elimination and serve as the basis for T cell-based immunotherapies. Here, we present an in vitro system that models key steps of antigen-specific T cell activation and cancer cell killing. Using OT-1 and P14 T cell receptor (TCR) transgenic mouse strains, we generate cytotoxic T lymphocytes (CTLs) through in vitro priming with cognate peptides. Cancer cells are engineered to present antigens via lentiviral transduction or peptide pulsing, facilitating the formation of MHC class I-antigen complexes recognized by the TCRs. These antigen-expressing cancer cells are then co-cultured with CTLs to assess antigen-specific cytotoxicity. Cancer cell viability is quantified by assessing the metabolic activity of living cells, providing a precise measure of CTL killing efficiency. As both lentiviral transduction and peptide pulsing are broadly applicable to various cancer cell models, this platform offers a versatile tool for dissecting antigen-specific cancer cell-T cell interactions and evaluating immunotherapeutic strategies.