Monitoring Influenza A Virus Entry Using Quantitative Fluorescence Microscopy

Area of Science:

• Virology
• Cell Biology
• Microscopy

Background:

• Influenza A virus (IAV) poses a significant global health threat, necessitating urgent development of broad-spectrum antivirals.
• Inhibiting IAV host cell entry is a promising strategy to block viral replication and transcription early in infection.
• Understanding IAV entry mechanisms is key to identifying host factors as therapeutic targets.

Purpose of the Study:

• To describe fluorescence microscopy-based assays for monitoring sequential IAV entry steps.
• To provide methods for quantifying IAV entry steps using high-content microscopy.
• To facilitate the discovery of novel antivirals and host-directed therapeutic interventions.

Main Methods:

• Monitoring IAV entry from cell surface attachment to genome delivery to the nucleus using fluorescence microscopy.
• Quantifying IAV entry steps via image analysis from high-content microscopy.
• Developing quantitative assays for high-content genetic or inhibitor screens.

Main Results:

• Established fluorescence microscopy assays to track IAV entry at sequential steps.
• Developed image quantification methods for major IAV entry stages.
• Provided tools to accelerate the identification of antivirals and host targets.

Conclusions:

• Fluorescence microscopy-based IAV entry assays enhance understanding of virus-host interactions.
• These methods can lead to the discovery of novel host-directed prophylactic or therapeutic interventions against influenza.
• The described assays support high-content screening for antiviral drug discovery.