The field of cancer immunotherapy has been revolutionized by the discovery of neoantigens, which are tumor-specific antigens that arise from somatic mutations in cancer cells. These neoantigens can be recognized by the immune system as foreign and trigger an immune response against the cancer cells. However, not all neoantigens are immunogenic, and predicting which ones are likely to elicit an immune response is a crucial step in developing effective cancer vaccines and immunotherapies. An immunogenic neoantigen must meet two or more requirements, the main bottleneck being proper MHC molecule presentation and efficient TCR recognition. According to recent studies, most neoantigens predicted by MHC molecular presentation do not elicit an immune response. Therefore, it is critical to consider TCR recognition by pMHC complexes when assessing the immunogenicity of potential neoantigens. The reactivity of neoantigens to T cells is mainly verified or screened by T cell-based assays, multicolor-labeled MHC tetramers, enzyme-linked immunosorbent spots (ELISpot) and T cell profiling.

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