Approximately 170 million are infected with the hepatitis C virus (HCV) world wide and in the United States an estimated 2.7 million are HCV RNA positive. Of those infected with HCV, 54-86% fail to clear the virus and develop a chronic infection. The chronic phase can last many decades and can ultimately lead to end stage liver disease. The efficiency of the current therapy for a HCV genotype 1 infection, the dominate genotype in the United States, is a 30% sustained viral response (SVR) rate To develop more efficient drugs and therapies, a better understanding of the HCV life cycle is required. For this project we focused our efforts on the initial phase of the HCV life cycle, the process of viral entry. First, we attempted to utilize biophysical assays to identify peptide inhibitors of HCV entry, a method previously used successfully in our laboratory for the SARS coronavirus. Second, we set about measuring the kinetics of a successful HCV cell culture strain (HCVcc) infection of naive Huh7 cells and how it relates to viral entry and decay. Finally, we examined the role viral particle density has on early infection kinetics of HCVcc We were able to identify four peptide inhibitors of HCV pseudo particle (HCVpp) infectivity where two of the peptides showed activity in our biophysical assays indicating that in the case of HCV, utilization of biophysical assays is not an efficient method to identify peptide inhibitors of HCV. Our examination of HCVcc infection kinetics, in vitro, revealed that HCVcc early infection kinetics is a slow process requiring 24 hours to reach completion with particle decay being a significant factor in deciding final infectious titers. When we studied the relationship between particle density and HCVcc early infection kinetics, we did not find a difference in early infection kinetics between different density fractions of HCVcc. We did, however, observe that during the process of infection the density profile of the viral inoculum shifts dramatically toward lighter density. We observed the same effect during HCVcc decay process as the density profile shifted toward lower densities as a function of time spent at 37°C / acase@tulane.edu
| Identifer | oai:union.ndltd.org:TULANE/oai:http://digitallibrary.tulane.edu/:tulane_27473 |
| Date | January 2008 |
| Contributors | Sabahi, Ali (Author), Flemington, Erik (Thesis advisor) |
| Publisher | Tulane University |
| Source Sets | Tulane University |
| Language | English |
| Detected Language | English |
| Rights | Access requires a license to the Dissertations and Theses (ProQuest) database., Copyright is in accordance with U.S. Copyright law |
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