Cellular characterization of antibodies raised against PA and PB2 of H1N1 virus

• Main Author: Lai, Nicole Poh Yee
• Other Authors: Richard J Sugrue
• Format: Final Year Project
• Language: English
• Published: Nanyang Technological University 2023
• Subjects: Science::Biological sciences
• Online Access: https://hdl.handle.net/10356/169887
• Institution: Nanyang Technological University

Influenza A virus is responsible for the global seasonal flu and pandemics that occur. Understanding spatial activity and interactions of influenza viral proteins and their contributions towards viral pathogenicity becomes pivotal in developing anti-viral strategies. Imaging is a powerful technique to assess intracellular protein expression and distribution. Immunofluorescence has the advantages of target specificity and signal amplification. Therefore, antibodies raised against PA and PB2 and other viral proteins were used to study their intracellular localizations within A549 cells. PA exhibited nuclear and cytoplasmic staining while PB2 showed only nuclear staining. Co-staining of different combinations of antibodies revealed colocalizations of PA and PB2 with M1 and NEP, as well as M1 and NEP with NP that support a proposed nuclear export model. Generally, these data suggested effective protein recognition by the antibodies of choice and can be used to monitor viral proteins during infection. Our results formed the basis for further investigations to confirm interactions between colocalizing proteins in cells more representative of the upper respiratory airway, such as nasal epithelial cells, to validate proposed models and elucidate mechanisms that could be targeted for therapeutic intervention.