Herpes simplex viruses (HSVs) are enveloped DNA viruses that cause human disease, including cold sores, eye and genital infections and encephalitis. In addition, genital herpes may play a role in the acquisition of AIDS. The virion envelope contains seven glycoproteins, which play important roles in virus infection, as targets of the immune response of the host, and pathogenesis. The long term objective of this grant is to understand the 3-dimensional structure of two essential HSV glycoproteins, gD and gH.
Aim 1 proposes studies of the antigenic and structural properties of gD, a protein involved in virus penetration, and a human subunit vaccine candidate. We have combined biochemical, immunologic and recombinant DNA techniques to address questions about gD structure, including a more precise understanding of antigenic epitopes, the position of disulfide bonds, the function of N-linked oligosaccharides, and the oligomeric state of gD. In order to interpret the accumulated data, the actual 3D structure of gD must be known. Therefore, experiments in Aim 2 are designed to isolate a crystallizable form of gD for 3-d structure determination by X-ray crystallography. Such studies would clarify the structural elements of gD involved in its functions. The strategy will be to obtain a crystallizable fragment by proteolysis of gD Aim 3 proposes studies of the antigenic and structural properties of gH, a protein involved in viral entry and cell-to -cell spread. Immunologic reagents will be prepared against the purified protein, and used to construct a detailed antigenic map. Mutations will be engineered into the cloned gH-1 gene, and mutant proteins will be expressed in transfected cells. The mutants will be used for epitope mapping, and to address other questions such as processing, transport and function. In addition we will use the cloned gH-1 gene to look for a gH-2 homolog.
Aim 4 proposes a study of the potential of HSV glycoproteins to function as subunit vaccine preparations and to answer the following question. 1) What effect do mutations have on the efficacy of gD as a vaccine? 2) Can other HSV glycoproteins protect animals? 3) Do other HSV glycoproteins enhance the protective capacity of gD?
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