Identification and Relative Abundance of Naturally Presented and Cross-reactive Influenza A virus MHC class I-restricted T Cell Epitopes

Cytotoxic T lymphocytes are key for controlling viral infection. Unraveling CD8+ T cell-mediated immunity to distinct influenza virus strains and subtypes across prominent HLA types is relevant for combating seasonal infections and emerging new variants. Using an immunopeptidomics approach, naturally presented influenza A virus-derived ligands restricted to HLA-A*24:02, HLA-A*68:01, HLA-B*07:02, and HLA-B*51:01 molecules were identified. Functional characterization revealed multifunctional memory CD8+ T cell responses for nine out of sixteen peptides. Peptide presentation kinetics was optimal around 12 hours post infection and presentation of immunodominant epitopes shortly after infection was not always persistent. Assessment of immunogenic epitopes revealed that they are highly conserved across the major zoonotic reservoirs and may contained single substitution in vicinity of the anchor residues. These findings demonstrate how the identified epitopes promote T cell pools possibly cross-protective in individuals and can be potential targets for vaccination.