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Elucidation of the effects of wild type Hepatitis C Virus on host signaling cascades
Aguado, Lauren Colette
Wolkowicz, RolandBernstein, SanfordHuxford, Tom
vii, 35 p. : ill.
Hepatitis C affects over 170 million people worldwide, with a high incidence of chronic infection that potentially leading to liver fibrosis and hepatocellular carcinoma. The Hepatitis C virus (HCV) is a single stranded positive sense RNA virus that is translated into four structural and six non-structural proteins. Many of the viral proteins have been shown independently to interact with and influence a number of proteins involved in host signaling cascades. These effects have yet to be discerned in the context of wild type viral infection. Our studies aims to shed light on how HCV infection induces alterations in host signaling cascades in the hepatocyte Huh 7.5.1 cell line. For that purpose, we use the Japanese Fluminant Hepatitis Virus-1 (JFH-1), a genotype 2a strain of HCV, recently shown to replicate in tissue culture. In order to elucidate the effects of viral infection on signaling, we exploit the power of PhosphoFlow, a flow cytometry-based technique that allows detection of phosporylation events through intracellular staining. Through the use of this technique, we observed activation of the p38 MAP kinase, ERK, and NF-[lower case kappa]B cascades at five days post infection. This increase likely correlates with an increase in viral proteins and interactions of those viral proteins with host signaling proteins. In addition to those findings, we have observed via confocal microscopy, nuclear localization of p-I[lower case kappa]B[lower case alpha] at five days post infection, much in contrast to its typical fate: proteosomal degradation. We further hypothesize that one or more viral proteins could mediate this altered localization as well. Elucidation of viral-host crosstalk at the molecular level can facilitate a better understanding of HCV infection and pathogenesis.
Includes bibliographical references (p. 28-33)
Master of Science (M.S.) San Diego State University, 2011
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