Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus

Balaji Manicassamy, Santhakumar Manicassamy, Alan Belicha-Villanueva, Giuseppe Pisanelli, Bali Pulendran, Adolfo García-Sastre

Research output: Contribution to journalArticle

260 Scopus citations

Abstract

Influenza A virus is being extensively studied because of its major impact on human and animal health. However, the dynamics of influenza virus infection and the cell types infected in vivo are poorly understood. These characteristics are challenging to determine, partly because there is no efficient replication-competent virus expressing an easily traceable reporter gene. Here, we report the generation of a recombinant influenza virus carrying a GFP reporter gene in the NS segment (NS1-GFP virus). Although attenuated when compared with wild-type virus, the NS1-GFP virus replicates efficiently in murine lungs and shows pathogenicity in mice. Using whole-organ imaging and flow cytometry, we have tracked the dynamics of influenza virus infection progression in mice. Imaging of murine lungs shows that infection starts in the respiratory tract in areas close to large conducting airways and later spreads to deeper sections of the lungs. In addition to epithelial cells, we found GFP-positive antigen-presenting cells, such as CD11b+CD11c -, CD11b-CD11c+, and CD11b +CD11c+, as early as 24 h after intranasal infection. In addition, a significant proportion of NK and B cells were GFP positive, suggesting active infection of these cells. We next tested the effects of the influenza virus inhibitors oseltamivir and amantadine on the kinetics of in vivo infection progression. Treatment with oseltamivir dramatically reduced influenza infection in all cell types, whereas, surprisingly, amantadine treatment more efficiently blocked infection in B and NK cells. Our results demonstrate high levels of immune cells harboring influenza virus antigen during viral infection and cell-type - specific effects upon treatment with antiviral agents, opening additional avenues of research in the influenza virus field.

Original languageEnglish (US)
Pages (from-to)11531-11536
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number25
DOIs
StatePublished - Jun 22 2010

Keywords

  • Antivirals
  • Cell tropism
  • GFP virus
  • Pathogenesis
  • Recombinant influenza virus

ASJC Scopus subject areas

  • General

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