Publication Date
2012
Document Type
Thesis
Committee Members
Paula Bubulya (Committee Member), Mill Miller (Committee Chair), Carol Sabourin (Committee Member)
Degree Name
Master of Science (MS)
Abstract
Influenza A H5N1 has emerged as a potential to be the next pandemic influenza. Ferrets are a promising model of H5N1 infection because the disease progression is similar to that known in humans, however reagents to characterize infections in ferrets are few. We developed real-time PCR assays for the ferret cytokines IL-1β, IL-2, IL-4, IL-8, IL-12p40, TNF-α, IFN-α, IFN-β, IFN-γ, TGFB-β1, and TGFB-β2. Forty-eight ferrets were vaccinated with either one of three vaccine formulations (Vaccine 1, Vaccine 2, or Vaccine 3), or saline (0.9% NaCl), then challenged intranasally with a lethal dose of H5N1 strain A/Vietnam/1203/04. Cytokine assays were then performed on blood drawn pre-vaccination, 0, 3, 4, and 14 days post infection, and brain, lung, and liver samples from ferrets euthanized either 4 or 14 days post infection or when found moribund. Ferrets vaccinated with Vaccine 1 showed an increase in IL-2, IL-6, IL-12p40, and IFN-β, and a decrease in TNF-α in the blood, an increase in IL-1β and TNF-α in the brain, an increase of IFN-β and IFN-γ in the lung, and an increase of IFN-γ in the liver during the course of infection. Ferrets vaccinated with Vaccine 2 showed an increase in IL-2, IL-12p40, IFN-α, IFN-β, IFN-γ, and TGFB-β2, and a decrease in TNF-α in the blood, an increase of IL-1β in the brain, an increase of IFN-β and IFN-γ in the lung, and little variance of cytokine levels in the liver during the course of infection. Ferrets vaccinated with Vaccine 3 showed a decrease in IL-2, IL-6, IL-8, IL-12p40, and TNF-α in the blood, little variance of cytokine levels in the brain and liver, and an increase in IL-4 in the lung during the course of infection. Ferrets with the saline control showed an increase in IL-2, and a decrease in IL-12p40, TNF-α and IFN-β in the blood, an increase of IL-1β, IL-8 and TNF-α in the brain, an increase of IFN-β and IFN-γ in the lung, and an increase of TNF-α and IFN-γ in the liver during the course of infection. Vaccine 3 conferred total protection, as indicated by 100% survival of the ferrets in that group. Vaccines 1 and 2 conferred some protection, with 22% and 33% of ferrets, respectively surviving. This study exhibits a promising vaccine in Vaccine 3 as well a new reagents for characterizing ferret immune responses, which may be useful in evaluating potential vaccine or therapeutic efficacy.
Page Count
104
Department or Program
Department of Biological Sciences
Year Degree Awarded
2012
Copyright
Copyright 2012, all rights reserved. This open access ETD is published by Wright State University and OhioLINK.
