We will apply newly available techniques to the study of the structure of the rabies virion, its replicative processes, and the disease that it causes. Because of the worldwide need for an economical and efficient rabies vaccine, we will attempt to develop a """"""""sub-unit"""""""" vaccine using recombinant DNA technology. We propose to clone a DNA sequence complementary to the mRNA of the rabies virus glycoprotein in an appropriate host vector system and to maximize its expression. The ideal immunogenic polypeptide for vaccine use will be determined by characterization of the critical protective antigenic site using specific monoclonal antibodies. The specificity of this site differs in viruses isolated in different regions of the world; present vaccines will not protect against all street viruses. We will compare the antigenic composition of representative street and fixed viruses in order to develop a vaccine of universally protective antigenic composition. We intend to perform a comparative analysis of genomic and subgenomic RNA structures, and to map the genomes of standard, phenotypically variant, and defective viruses, coupled with characterization of their protein antigenic structure using monoclonal antibodies. We will investigate the replication of standard and defective-interfering viruses in acute and chronic infections in cell culture (including differentiated and non-differentiated neuroblastoma and myoblast cells) and in vivo, using nucleic acid and monoclonal anitbody probes. Effects of rabies virus infection on cell energy metabolism will be studied. The role of antibody to rabies virus and of the cell-mediated immune response, including the specific trafficking of isolated subsets of immune cells into the CNS, will be further evaluated. We will seek to identify those components of the immune response that are either critical to recovery of the host, or, alternatively, contribute to the pathologic process.
| Bagasra, O; Michaels, F H; Zheng, Y M et al. (1995) Activation of the inducible form of nitric oxide synthase in the brains of patients with multiple sclerosis. Proc Natl Acad Sci U S A 92:12041-5 |
