T Cell Antigen Recognition and Discrimination by Electrochemiluminescence Imaging

Abstract Adoptive T lymphocyte (T cell) transfer and tumour‐specific peptide vaccines are innovative cancer therapies. An accurate assessment of the specific reactivity of T cell receptors (TCRs) to tumour antigens is required because of the high heterogeneity of tumour cells and the immunosuppressive tumour microenvironment. In this study, we report a label‐free electrochemiluminescence (ECL) imaging approach for recognising and discriminating between TCRs and tumour‐specific antigens by imaging the immune synapses of T cells. Various T cell stimuli, including agonistic antibodies, auxiliary molecules, and tumour‐specific antigens, were modified on the electrode's surface to allow for their interaction with T cells bearing different TCRs. The formation of immune synapses activated by specific stimuli produced a negative (shadow) ECL image, from which T cell antigen recognition and discrimination were evaluated by analysing the spreading area and the recognition intensity of T cells. This approach provides an easy way to assess TCR‐antigen specificity and screen both of them for immunotherapies.