Oral Presentation Australia and New Zealand Society for Extracellular Vesicles Conference 2025

Hijacking Clearance: How Influenza A Virus Exploits Apoptotic Bodies to Evade Phagolysosomal Neutralization (127307)

Stephanie F Rutter 1 , Georgia K Atkin-Smith 2 , Ivan K. H Poon 1
  1. La Trobe Insistute of Molecular Science, Bundoora, VIC, Australia
  2. Walter and Eliza Hall Institute, Parkville

Apoptosis and the clearance of apoptotic cells are essential to maintain physiological homeostasis.

Our previous work identified a series of protein regulators that govern the efficient dismantling of apoptotic cells into large EVs known apoptotic bodies (ApoBDs). These ApoBDs can communicate phagocytes through “find-me” and “goodbye” signals, and receptor-mediated interactions, such as phosphatidylserine-binding. Notably, we demonstrated that apoptotic cell disassembly plays a key role in host-pathogen interactions whereby Influenza A virus (IAV) and SARS-COV-2 viruses can induce fragmentation, resulting in ApoBDs capable of propagating viral infection in vitro and in vivo. Unexpectedly, recent work examining the mechanism of ApoBD-mediated viral transfer revealed that, in SARS-CoV-2, propagation of infection is efferocytosis-dependent, indicating that the virus can escape phagolysosome-mediated neutralization. However, if this mechanism is unique to SARS-COV-2 or is conserved across other respiratory viruses remains unknown.

 Here, we employed FACs-based isolation to collect “engulfing” and “non-engulfing” phagocytes following co-incubation with ApoBDs generated during IAV viral infection. Through confocal microscopy and flow cytometry, we observed that “engulfing” phagocytes exhibited a significant enrichment of IAV proteins compared to “non-engulfing” phagocytes, suggesting that IAV propagation may also be efferocytosis-dependent. Importantly, these data highlight that efferocytosis of IAV-infected apoptotic material is not always abortive. To investigate how IAV escapes phagolysosomal neutralization, we developed a pharmacological screening approach using commercially available antivirals. Overall, these findings identify a novel mechanism of IAV infection mediated by ApoBDs and enhance our understanding of the role of ApoBDs and dying cell clearance in host-pathogen interactions.