Varicella-zoster virus (VZV) is the human herpesvirus which is the etiologic agent of both the common childhood illness chickenpox and, upon reactivation, the dermatomal exanthem shingles (herpes zoster). Although only moderately distressful in healthy people, both diseases are important causes of morbidity and mortality in children and young adults with cancer or other chronic illnesses being treated with intensive chemotherapy or bone marrow/organ transplantation. Because of the serious sequelae of VZV infection in immunocompromised patients, the NIH-NIAID is currently sponsoring a collaborative clinical trial of a live attenuated VZV vaccine. The goal of this grant proposal is to define and characterize major VZV antigenic determinants. Glycoproteins specified by VZV will be given the highest priority since studies of other enveloped viruses demonstrate that glycosylated antigens play an important role in the induction of protective humoral and cell-mediated immune responses by the infected host; the same glycoproteins also mediate early events in the interaction between virus and cell.
The specific aims i nclude the following investigations: (1) produce panels of murine monoclonal antibodies to glycoproteins specified by a laboratory and a vaccine VZV strain, (2) identify the precursor and mature VZV glycosylated constituents of the infected cell and the enveloped virion, (3) study the synthesis, processing and intracellular transport of the viral glycoproteins, (4) isolate and purify the individual viral glycoproteins and perform structural analysis of the protein and sugar moieties, and (5) correlate biologic functions, e.g., neutralization, antibody dependent cell mediated cytotoxicity and delayed type hypersensitivity with determinants on individual glycoproteins. Techniques will include radioimmune precipitation, SDS-PAGE, immunoblotting, affinity and high performance liquid chromatography, tryptic peptide mapping, Bio-Gel filtration of glycopeptides, methylation analyses of carbohydrates and immunoelectron microscopy. This research project, therefore, will examine the biology, the biochemistry, and the immunobiochemistry of the VZV glycoproteins. In a collaborative research endeavor, the structural genes which encode the individual viral glycoproteins will be mapped by recombinant DNA technology.
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