Chicken intestinal organoids reveal polarity-dependent replication dynamics and immune responses of low pathogenic...

Low pathogenic avian influenza viruses (LPAIVs) persist in poultry populations, posing an ongoing challenge to poultry management and research. These viruses typically cause mild infections but can lead to significant economic losses due to their widespread presence and potential to disrupt poultry production. Traditional in vivo and in vitro models struggle to accurately replicate the avian intestinal environment, where these viruses often establish infection. In Taiwan, the domestic H6N1 LPAIVs cause an endemic in the local area but still lack investigation. This study addresses this gap by utilizing advanced chicken intestinal organoid (CIO) systems, apical-out (Ap-o), and basal-out (Ba-o) conformations to study the unique replication kinetics and innate immune responses of LPAIVs in a physiologically relevant setting. By comparing the Taiwan specialized H6N1 strain toward the Eurasian H9N2 virus, our results demonstrate that Ap-o organoids, which mimic natural exposure to the intestinal lumen, elicit robust interferon-stimulated gene responses, particularly higher expression of downstream gene, which effectively controls viral replication against H6N1 virus. In contrast, Ba-o organoids, representing a systemic infection route, exhibited lower upstream interferon responses, reflecting a different immune response pattern in the H9N2 strain. These results confirm that CIO is a well-suited model to study LPAIV pathogenesis. It provides key insights into the host-pathogen interactions that determine viral replication and immune evasion strategies. This model deepens our understanding of LPAIV behavior in poultry and provides a valuable tool for developing more targeted and effective control strategies for poultry health management.