High-throughput and high-dimensional single cell profiling for antigen-specific CD4+ T cells 2757

Abstract Description Although identification and characterization of antigen-specific CD4+ T cells remains crucial for understanding immune responses, several technique hurdles have made it challenging to do so. Building upon our TetTCR-SeqHD platform, we developed a high-throughput and high-dimensional profiling technology, TetTCRII-SeqHD, for antigen-specific CD4+ T cells. The technology leverages the recently developed affinity enhanced class II MHC tetramer and optimizes the staining conditions to address the inherently lower TCR-pMHC class II binding avidity. Multiple tetramers can be multiplexed to simultaneously track T cells with different antigen specificities. Validation using multiple TCRs with known antigens demonstrated an average of > 90% precision and >80% recall rate, comparable to conventional methods while providing comprehensive transcriptional and phenotypic information at single-cell resolution. This advancement of TetTCRII-SeqHD provides valuable insights into helper T cell responses with broad applications in studying infection, vaccination, autoimmune diseases, and cancer immunotherapy. Funding Sources Supported by NIH U19AI128949 and R33CA256086. Topic Categories Technological Innovations in Immunology (TECH)