A universal MHCII technology platform to characterize antigen-specific CD4+ T cells

Abstract

CD4+ Tcells are critical to the immune system and perform multiple functions; therefore, their identification and characterization are crucial to better understanding the immune system in both health and disease states. However, current methods rarely preserve their exvivo phenotype, thus limiting our understanding of their invivo functions. Here we introduce a flexible, rapid, and robust platform for exvivo CD4+ Tcell identification. By combining MHCII allele purification, allele-independent peptide loading, and multiplexed flow cytometry technologies, we can enable high-throughput personalized CD4+ Tcell identification, immunophenotyping, and sorting. Using this platform in combination with single-cell sorting and multimodal analyses, we identified and characterized antigen-specific CD4+ Tcells relevant to COVID-19 and cancer neoantigen immunotherapy. Overall, our platform can be used to detect and characterize CD4+ Tcells across multiple diseases, with potential to guide CD4+ Tcell epitope design for any disease-specific immunization strategy.