The long term goals of this research project are: 1) the isolation and in vitro construction of Sindbis virus mutants which differ in their pathogenic effects on experimental animals, 2) the detailed characterization of such mutants as to genome sequence and virion topography and correlation of specific mutations with their biological and pathogenesis phenotypes, and 3) the use of this model system to explore informed molecular approaches to the development of live alphavirus vaccines. The ability to construct in vitro recombinant viruses and generate site-directed mutations is one of two considerations which make the Sindbis system an excellent model for the detailed examination of the molecular basis of alphavirus neurovirulence. The second consideration is that the prior mapping of pathogenesis loci in biologically selected mutants by our group will guide the targeting of site-directed mutagenesis to specific regions. At this time, few other virus systems have both of these elements. The experiments proposed in this application will explore the molecular genetics of Sindbis virus pathogenesis in mice. Specific loci which affect pathogenesis will be mapped and characterized utilizing closely related, biologically selected mutants as well as site-directed mutagenesis of a full- length cDNA clone of the viral genome. The effects of these mutations will be assessed in animal models and in cell culture systems which display inducible differentiation toward neuron-like cells. Knowledge of the molecular genetics of Sindbis pathogenesis will be extended by the informed design of a low-reversion, model vaccine containing multiple, independently attenuating mutations. Finally, while the focus of the proposed experiments is the molecular genetics of alphavirus pathogenesis, the results obtained will be important to the understanding of penetration, virus maturation and structure, and post-translational processing by proteolytic cleavage. The proposed investigation will yield significant new information regarding the pathogenesis and replication of the alphavirus group.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI022186-06
Application #
3133000
Study Section
Virology Study Section (VR)
Project Start
1985-12-01
Project End
1994-01-31
Budget Start
1991-02-01
Budget End
1992-01-31
Support Year
6
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Type
Schools of Medicine
DUNS #
078861598
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
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Ryman, Kate D; Gardner, Christina L; Burke, Crystal W et al. (2007) Heparan sulfate binding can contribute to the neurovirulence of neuroadapted and nonneuroadapted Sindbis viruses. J Virol 81:3563-73
Ossiboff, Robert J; Sheh, Alexander; Shotton, Justine et al. (2007) Feline caliciviruses (FCVs) isolated from cats with virulent systemic disease possess in vitro phenotypes distinct from those of other FCV isolates. J Gen Virol 88:506-17
Klimstra, William B; Williams, Jacqueline C; Ryman, Kate D et al. (2005) Targeting Sindbis virus-based vectors to Fc receptor-positive cell types. Virology 338:9-21

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