Generating Influenza Pseudotyped Viral Particles

The IAV subtypes used in this study, including the pH1N1 (A/England/2009) and H3N2 (A/HK/1999), were provided by Wendy Barclay (Imperial College, London) and Leo Poon (University of Hong Kong), respectively. The plasmids used for the production of H1N1 and H3N2 pseudo-typed viral particles were obtained from Addgene; pHIV-Luciferase (Addgene plasmid #21375); psPAX2 (Addgene plasmid # 12260); and Vesicular Stomatitis Virus (VSV-G) (Addgene plasmid #8454). pcDNA3.1-swineH1-flag (H1 from swine H1N1 A/California/04/09) (Invitrogen), pcDNA3.1-swine N1-flag (N1 from swine H1N1 A/California/04/09), and pCDNA H3 (from A/Denmark/70/03 (H3N2) (Invitrogen) were obtained commercially. pI.18-N2 [N2 from A/Texas/50/2012/(H3N2)] plasmid was a gift from Nigel Temperton (University of Kent). Anti-influenza antibodies used were obtained from BEI Resources, NIAID, NIH, USA, and used as previously described (38 (link)); these include polyclonal anti-influenza Virus H3 HA, A/Hong Kong/1/1968 and monoclonal anti-influenza virus H1 HA, A/California /04/2009.

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Murugaiah V., Varghese P.M., Saleh S.M., Tsolaki A.G., Alrokayan S.H., Khan H.A., Collison K.S., Sim R.B., Nal B., Al-Mohanna F.A, & Kishore U. (2020). Complement-Independent Modulation of Influenza A Virus Infection by Factor H. Frontiers in Immunology, 11, 355.

Publication 2020

pHIV-Luciferase psPAX2 pcDNA3.1-swineH1-flag pcDNA3.1-swine N1-flag pCDNA H3

Anti antibodies Influenza Influenza virus Luciferase Plasmid Swine Vesicular stomatitis virus Viral particles

Corresponding Organization :

Other organizations : Brunel University of London, Vellore Institute of Technology University, King Faisal Specialist Hospital & Research Centre, King Saud University, University of Oxford

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Variable analysis

independent variables

IAV subtypes used in this study, including the pH1N1 (A/England/2009) and H3N2 (A/HK/1999)
Plasmids used for the production of H1N1 and H3N2 pseudo-typed viral particles (pHIV-Luciferase, psPAX2, and Vesicular Stomatitis Virus (VSV-G))
Commercially obtained plasmids (pcDNA3.1-swineH1-flag, pcDNA3.1-swine N1-flag, and pCDNA H3)
PI.18-N2 [N2 from A/Texas/50/2012/(H3N2)] plasmid

dependent variables

Measured outcomes (not explicitly mentioned)

control variables

Anti-influenza antibodies used (polyclonal anti-influenza Virus H3 HA, A/Hong Kong/1/1968 and monoclonal anti-influenza virus H1 HA, A/California /04/2009)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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