Viral lineage and mode of exposure modulate within host spatial dynamics of influenza A viruses

Leyson, C.; Vargas-Maldonado, N.; Gaddy, M.; Raghunathan, V.; Matias-Ferreri, L.; Sethi, M.; Patatanian, K.; Carnaccini, S.; Ganti, K.; VanInsberghe, D.; Lowen, A. C.

doi:10.1101/2025.11.03.686270

bioRxiv · 10.1101/2025.11.03.686270

Viral lineage and mode of exposure modulate within host spatial dynamics of influenza A viruses

Leyson, C.·Vargas-Maldonado, N.·Gaddy, M.·Raghunathan, V.·Matias-Ferreri, L.·Sethi, M.·Patatanian, K.·Carnaccini, S.·Ganti, K.·VanInsberghe, D.·Lowen, A. C.

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Abstract

The upper and lower respiratory tracts (URT and LRT) present distinct environments for influenza A virus (IAV) replication. Their differential features have major implications for viral evolutionary dynamics, transmission potential, and pathogenesis. To investigate the implications of differential viral replication in the URT and LRT, we assessed dispersal of IAVs throughout the guinea pig respiratory system. Guinea pigs were inoculated intranasally with a 300 L volume to deliver inoculum to both the URT and LRT. Two strains were used to represent the circulating seasonal IAV lineages: influenza A/TX/50/2012 (H3N2) and influenza A/CA/07/2009 (H1N1) virus. The inclusion of a diverse genetic barcode enabled high-resolution tracing of viral dispersal for the H1N1 virus. While infectious virus was consistently detected in the URT, the H1N1 virus could be detected in LRT while the H3N2 virus could not. To determine whether replication of the H1N1 virus in the LRT extends to other modes of infection, virus distribution was evaluated following infection via aerosol exposure or transmission. Infectious virus in lung homogenates was observed in both cases, confirming the LRT tropism of the H1N1 virus. Sequencing genetic barcodes revealed that diversity was largely maintained in nasal samples and trachea but contracted upon dispersal to the lungs. This loss of diversity was associated with increased distance to and branching from the major airways, implicating long distance dispersal through the airways in imposing within-host population bottlenecks. These data underline the implications for within-host viral dynamics of the distinct environments of the upper and lower respiratory tracts. ImportanceThe upper (URT) and lower (LRT) respiratory tracts create different conditions for influenza A virus (IAV) spread and evolution. We studied how the virus moves through guinea pigs airways after infection with H3N2 or H1N1 strains of IAV. Whether delivered intranasally, by aerosol or by transmission, the H1N1 virus replicated in the nasal cavity, trachea, and lungs. By contrast, the H3N2 virus stayed mostly in the nasal cavity. Genetic barcodes were used to track how the H1N1 virus moved and changed. The populations replicating in the nasal cavity and trachea maintained high diversity but those sampled from the lungs showed low diversity. This bottlenecking effect was stronger for viral populations present deeper in the lungs. These findings show that the different environments of the URT and LRT strongly shape how influenza spreads and evolves inside a host.

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Leyson, C., Vargas-Maldonado, N., Gaddy, M., Raghunathan, V., Matias-Ferreri, L., Sethi, M., Patatanian, K., Carnaccini, S., Ganti, K., VanInsberghe, D., Lowen, A. C.. 2025-11-03. Viral lineage and mode of exposure modulate within host spatial dynamics of influenza A viruses. https://doi.org/10.1101/2025.11.03.686270

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