Abstract

CD8+ T cells are essential effectors in antiviral immunity, recognizing short virus-derived peptides presented by MHC class I (pMHCI) on the surface of infected cells. However, the fraction of viral pMHCI on infected cells that are immunogenic has not been shown for any virus. To approach this fundamental question, we used peptide sequencing by high-resolution mass spectrometry to identify more than 170 vaccinia virus pMHCI presented on infected mouse cells. Next, we screened each peptide for immunogenicity in multiple virus-infected mice, revealing a wide range of immunogenicities. A surprisingly high fraction (>80%) of pMHCI were immunogenic in at least one infected mouse, and nearly 40% were immunogenic across more than half of the mice screened. Further, we have extended these findings to track the kinetics of epitope presentation during infection of dendritic and fibroblast cell lines, as well from splenocytes from infected mice. The high number of peptides found to be immunogenic and the distribution of responses across mice—in addition to comprehensive profiling of presentation kinetics and binding affinity for MHCI—give us insight into the specificity of antiviral CD8+ T cell responses.

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