The candidate is a pediatrician who has completed her clinical training in pediatric infectious diseases. Her goal is to pursue an academic career in this subspeciality. Her preliminary research experience has allowed her to identify viral immunology as her general area of research interest. She seeks a Clinical Investigator Award for three years of support in order to acquire the necessary theoretical background and laboratory experience to become a competent independent investigator. She has designed a didactic program and a proposal for supervised laboratory research under the sponsorship of Dr. Bernard Roizman, Molecular Genetics and Cell Biology, and Dr. Jeff Bluestone, Pathology. The research proposal focuses upon the T-lymphocyte immunity to herpes simplex virus in the mouse model. Herpes simplex virus causes serious disease in previously healthy and immunocompromised patients. There is evidence that T-lymphocyte immunity plays a major role in the immunity to herpes viruses in humans during the recovery from acute infection. The role that specific viral proteins play in eliciting cytotoxic T-cells will be studied. As components of the viral envelope, HSV glycoproteins are likely to be important in inducing immunity. The purpose of this proposal is to examine the interaction of cytotoxic T-lymphocytes with HSV-1 glycoprotein E (gE), glycoprotein I (gI), and the glycoprotein E/glycoprotein I (gE/gI) complex in the murine model of HSV-1 infection and to correlate the host response to these proteins with disease pathogenesis. The proposal consists of two major phases. In the first phase, gE and gI will be cloned and then transfected into a major histocompatibility complex (MHC) matched murine fibroblast cell line which expresses both class I and II MHC proteins. These constructed cell lines will be used in the second phase of the project as stimulating antigen for the cloning of HSV specific T-cells and targets in the cytotoxicity assays. CBA mice will be infected by footpad injection with HSV-1 and after six days their draining popliteal lymph nodes will be isolated and cultured in vitro. Cultures exhibiting cytolytic capabilities will be characterized as to their phenotypic markers (i.e. CD3, CD4, CD8, MHC-1, MHC-II). T-cell surface antigens will be studied by fluorescence activated cell sorting analysis. Adoptive transfer experiments with protein specific cloned T-cells will provide a measure of the protective efficacy of gE, gI, or gE/gI CTL given during acute infection. These studies will help to delineate the effect that gE and gI have upon the immune system and whether their inclusion in any HSV vaccine would be beneficial or detrimental to the host.
