MHC-II acts as a fusion-triggering receptor for bat influenza virus

Influenza A virus hemagglutinin is a prototypical class I viral fusion protein that binds sialylated glycans and is activated by low pH in endosomes. In contrast, bat-derived IAV subtypes H17N10 and H18N11 use major histocompatibility complex class II (MHC-II) as an entry receptor, but how this receptor contributes to membrane fusion remains unknown. We find that MHC-II-dependent hemagglutinin subtypes H17, H18, and H19 possess an increased negative net charge relative to canonical HAs. Using cryo-electron tomography, we demonstrate that H18N11 morphology remains stable and H18 is in prefusion conformation at strongly acidic pH. Remarkably, H18 undergoes fusion-relevant conformational changes only when both MHC-II binding and low pH are present. By reconstitution of H18N11 fusion with liposomes and purified MHC-II, we show that receptor engagement is required to trigger the fusion activity of H18. These findings identify MHC-II as a receptor that directly triggers membrane fusion and reveal a previously unrecognized receptor-dependent mechanism of influenza virus entry. SignificanceBat-derived influenza A viruses (IAV) challenged established paradigms because they use major histocompatibility complex class II (MHC-II) molecules instead of sialylated glycans for cell entry. Recent studies suggest that MHC-II usage extends beyond bats, with avian, swine, and human strains showing dual receptor specificity. However, how MHC-II contributes to membrane fusion has remained unclear. We show that, unlike conventional HAs that are activated by acidity alone, the H18 fusion protein requires receptor engagement and low pH to undergo fusion-relevant conformational changes leading to membrane fusion. Hence, we reveal a novel receptor-dependent mechanism of membrane fusion for the MHC-II dependent entry of H18N11.