LAUSR

Significance Immunotherapy is a promising approach to treat cancers, infectious diseases, and autoimmunity by harnessing the power of immune cells, especially T cells. To improve the precision and efficacy of immunotherapy, the ability to genetically modify antigen-specific T cells is needed but cannot be accomplished using current methods. Here, we present a method, V-CARMA (Viral ChimAeric Receptor MHC-Antigen), to generate lentiviruses displaying peptide-MHC complex to specifically target T cells that express cognate TCRs and subsequently deliver genes into target T cells for genetic modification. Our results demonstrate that V-CARMA is a versatile tool to detect and modify antigen-specific T cells. T cells promote our body’s ability to battle cancers and infectious diseases but can act pathologically in autoimmunity. The recognition of peptides presented by major histocompatibility complex (pMHC) molecules by T cell receptors (TCRs) enables T cell–mediated responses. To modify disease-relevant T cells, new tools to genetically modify T cells and decode their antigen recognition are needed. Here, we present an approach using viruses pseudotyped with peptides loaded on MHC called V-CARMA (Viral ChimAeric Receptor MHC-Antigen) to specifically target T cells expressing cognate TCRs for antigen discovery and T cell engineering. We show that lentiviruses displaying antigens on human leukocyte antigen (HLA) class I and class II molecules can robustly infect CD8+ and CD4+ T cells expressing cognate TCRs, respectively. The infection rates of the pseudotyped lentiviruses (PLVs) are correlated with the binding affinity of the TCR to its cognate antigen. Furthermore, peptide-HLA pseudotyped lentivirus V-CARMA constructs can identify target cells from a mixed T cell population, suppress PD-1 expression on CD8+ T cells via PDCD1 shRNA delivery, and induce apoptosis in autoreactive CD4+ T cells. Thus, V-CARMA is a versatile tool for TCR ligand identification and selective T cell manipulation.