Toward Predicting Pandemic Potential: A Comparative Analysis of Virus-Host Interactions Between Diverse Influenza A Viruses and the Human Innate Immune System: Raw data from mN2H screening and functional studies

Zoonotic infections caused by avian influenza A viruses (IAVs) are a major public health concern, as they may represent the initial step toward the emergence of a pandemic strain. The host range of IAVs is shaped by species-specific virus–host interactions, and several host barriers have been identified that restrict the adaptation of avian viruses to humans. In this study, we used a comparative interaction screen by mammalian cell-based N2H (mN2H) assay based on the NanoLuc luciferase to investigate whether differences in the interaction networks between viral proteins from human, avian, and zoonotic IAVs and a curated panel of human innate immune factors could serve as indicators of the zoonotic potential of avian IAVs. The results revealed both conserved and strain-specific interactions, while underscoring limitations in comparing interactions across independent screens. Complementary functional analyses by siRNA knockdowns identified host factors and pathways potentially involved in host restriction, as well as pro-viral factors that could represent potential antiviral targets. The comparative interactomic mN2H approach holds promise for evaluating the zoonotic potential of avian IAVs, but our findings highlighted the need for further studies using broader cellular libraries to fully validate its predictive value.