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The Washington Research Foundation Fellowship

Nicholas Provine, Microbiology and Biology - 2009-10 WRFF

Nick Provine photoI am a graduating senior in the department of microbiology. I began my research career two and a half years ago in Dr. Nancy Haigwood’s lab working on HIV vaccine development. I joined Dr. Julie Overbaugh’s lab a year and a half ago when Dr. Haigwood moved to OHSU. When I joined Dr. Overbaugh’s lab I began my current project looking at different in vitro construction techniques of HIV. There are several widely used virus construction techniques. However, there has been very little direct comparison of these systems to confirm that results generated in each system are comparable. My project’s goal is to directly compare these systems so that results from other studies that use each of these systems can be compared. The experiences of working on an independent project have exposed me to countless opportunities and an absolute love of scientific research. Recently, my research gave me the chance to travel to the 16th Annual West Coast Retrovirus Conference in Palm Springs, CA to present my findings. Upon completing my undergraduate degree I plan to pursue a Ph.D. in HIV research. I am very grateful for receiving a Washington Research Foundation Fellowship, as it will allow me to continue my research during my senior year and complete my project before I graduate.

Mentor: Julie Overbaugh, Microbiology

Project Title: Phenotypic Characterization of Four Human Immunodeficiency Virus type 1 Model Systems

Abstract: Since human immunodeficiency virus type I (HIV-l) came under study over 25 years ago the goal of many vaccine attempts has been the elicitation of neutralizing antibodies (NAbs), which are widely believed to be the immunizing force behind most successful vaccines. Historically, the gold standard for in vitro testing of NAbs has been the peripheral blood mononuclear cell (PBMC) based assay. Recently the pseudovirus (PV) neutralization assay, which addresses the shortcomings of the PBMC assay, has been put forward as a widely adopted alternative. Previous studies have demonstrated that genetically identical virus display different neutralization phenotypes when tested side-by-side in these two assays. There are several major differences between the two experimental systems, and it remains unclear which of these variables accounts for the discrepancies. In my study I propose a systematic way to determine what variable (or combination of variables) accounts for the differences in the two systems. The three variables under study are the cell type the virus is produce in (either tissue culture or PBMC), the target cell of the assay (either tissue culture or PBMC), and whether the virus capable of just one or multiple rounds of infection of the target cells. To accomplish these goals I have developed a panel of 13 genetically matched PV and replication competent chimeras, which are representative of global HIV biodiversity. This panel will allow for each variable to be isolated while all others are held constant Preliminary results, in good agreement with previously published data, demonstrate that the replication competency of the virus is not a variable that affects susceptibility to NAbs. Over the course of the next year I will elucidate what role the other factors play in determining neutralization phenotype.