Validation of anti-ferret antibodies in an influenza infection model

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Presentation Time: 03:15 PM - 04:30 PM
Poster Board Number: B826
Abstract ID: 5445

Presenting Author:
Deborah M. Brown , Principal Investigator at Trudeau Inst., Clarkson Univ.

Abstract:

Ferrets represent excellent models of infectious disease due to their natural susceptibility to human respiratory viruses including influenza A virus (IAV), yet reagents to track immune responses are extremely inadequate. We developed a panel of monoclonal antibodies to better understand and characterize the immune response to respiratory infections in the ferret. We utilized the IAV infection model in which ferrets were infected with A/California/04/2009 H1N1 virus. We demonstrate that IAV titers peak at day (d) 2 in nasal wash samples and decrease over time until d7. Concurrent with viral kinetics, CCR2, CCR4 and CXCL10 peak at d2 and decrease to background levels by d10. Inflammatory cytokines IL-6 and TNF-a also peak at d2, show another increase at d7 before decreasing at d14. Antibodies to cell surface CD64, CD56 and CD86 were also developed and multi-color flow cytometry with existing cross-reactive antibodies demonstrate that CD64 (FcgRI) co-stains a population of cells that are positive for CD11b and CD14, but not CD4 or CD8 in uninfected spleen. Anti-ferret CD86 also co-localizes with CD64, CD11b and CD79, indicating these new reagents can be used to track macrophages and activated antigen presenting cells upon infection. Future work will track populations in IAV infected ferrets in flow cytometry and fluorescent microscopy. Together, utilization of these reagents will open new avenues of investigation for the immunology community in respiratory infection models.