Laura Cook

Activation‐induced marker (AIM) assays have proven to be an accessible and rapid means of antigen‐specific T‐cell detection. The method typically involves short‐term incubation of whole blood or peripheral blood mononuclear cells with antigens of interest, where autologous antigen‐presenting cells process and present peptides in complex with major histocompatibility complex (MHC) molecules. Recognition of peptide–MHC complexes by T‐cell receptors then induces upregulation of activation markers on the T cells that can be detected by flow cytometry. In this review, we highlight the most widely used activation markers for assays in the literature while identifying nuances and potential downfalls associated with the technique. We provide a summary of how AIM assays have been used in both discovery science and clinical studies, including studies of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) immunity. This review primarily focuses on AIM assays using human blood or peripheral blood mononuclear cell samples, with some considerations noted for tissue‐derived T cells and nonhuman samples. AIM assays are a powerful tool that enables detailed analysis of antigen‐specific T‐cell frequency, phenotype and function without needing to know the precise antigenic peptides and their MHC restriction elements, enabling a wider analysis of immunity generated following infection and/or vaccination.