Figure 5.

Cell-based TCR screening strategies. A) Target cells are labeled with radioactive chromium (⁵¹Cr) and co-cultured with cytotoxic T cells. Upon recognition and lysis of target cells by T cells, chromium is released into the supernatant. The level of chromium release is measured to assess T cell cytotoxicity, providing a quantitative evaluation of the T cell-mediated killing of cancer cells. B) T cells from patients or donors are co-cultured with target cells expressing a library of antigens. The proliferation of T cells is monitored by the incorporation of BrdU into newly synthesized DNA during cell division. The level of BrdU incorporation, detected through fluorescence microscopy, reflects the strength of the TCR-pMHC interaction, providing insights into T cell responses against specific antigens. C) ELISpot is used to detect cytokine secretion from individual T cells in response to target cells. T cells are co-cultured with target cells, and cytokine release is captured on a membrane pre-coated with specific antibodies. After incubation and washing, spots representing single T cell cytokine release are visualized. Flow cytometry-based sorting can then be used to isolate specific subsets of T cells for further analysis. D) In T-Scan, T cells are cultured with target cells expressing an antigen library. Granzyme B release upon target cell recognition is measured, serving as an indicator of cytotoxic activity. Target cells are sorted based on granzyme B reporter expression, and next-generation sequencing is used to identify the antigen that triggered the T cell response, enabling the identification of novel antigens. The images in the figures were created using BioRender (https://www.biorender.com/).