Venezuelan equine encephalitis virus (VEE) is a lymphotropic and neurotropic alphavirus, currently causing an explosive human epidemic in Colombia and Venezuela with over 45,000 cases to date. The long-term objectives of the proposed experiments are to define the molecular genetics of VEE induced disease and to use the VEE system to elucidate fundamental aspects of viral pathogenesis. The approach combines molecular genetics with classical histopathology and in situ hybridization. The experiments will utilize a mouse model which approximates VEE disease, molecularly cloned wild-type virus (V3000) which recapitulates the pathogenesis of natural virus isolates; and avirulent or highly attenuated site-directed mutants of V3000. Each of the mutations interdicts the VEE pathogenic sequence at a different stage and therefore links specific phenotypes in vivo with defined mutations. Vector systems have been developed which carry the gene for green fluorescent protein packaged in wild-type or mutant glycoprotein envelopes, facilitating the sensitive detection of ongoing virus replication in living cells or prior replication in preserved tissues. The proposed experiments take advantage of these powerful tools for further genetic dissection of VEE pathogenesis. The experiments are organized into four Specific Aims: 1) To define parameters of the lymphotrophic stage of VEE pathogenesis (e.g. elucidation of the earliest events in infection, identification of lymphoid target cells in vivo and in vitro, and the source of the viremia), 2) To determine the mechanism by which avirulent VEE mutants induce mucosal IgA following subcutaneous inoculation (e.g. characterization of viral spread to inductive sites of the mucosal immune system and correlation of mucosal IgA induction with virus mutation, mouse strain, and ability to spread to inductive sites), 3) To define parameters related to invasion of, dissemination within, and clearance of virus from the CNS (e.g. genetic characterization of CNS invasion, synaptic transmission, persistent infections in cell culture and animals, antibody curing of neuronal infections, and the mechanism of VEE induced neuronal death), and 4) To determine the in vivo and cell culture phenotypes of mutations at nucleotide 3 in the 5' untranslated region of the VEE genome.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS026681-12
Application #
6187208
Study Section
Experimental Virology Study Section (EVR)
Program Officer
Kerza-Kwiatecki, a P
Project Start
1989-03-01
Project End
2001-09-29
Budget Start
2000-05-01
Budget End
2001-09-29
Support Year
12
Fiscal Year
2000
Total Cost
$269,014
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Ryman, Kate D; White, Laura J; Johnston, Robert E et al. (2002) Effects of PKR/RNase L-dependent and alternative antiviral pathways on alphavirus replication and pathogenesis. Viral Immunol 15:53-76
Charles, P C; Trgovcich, J; Davis, N L et al. (2001) Immunopathogenesis and immune modulation of Venezuelan equine encephalitis virus-induced disease in the mouse. Virology 284:190-202
White, L J; Wang, J G; Davis, N L et al. (2001) Role of alpha/beta interferon in Venezuelan equine encephalitis virus pathogenesis: effect of an attenuating mutation in the 5' untranslated region. J Virol 75:3706-18
Schultz-Cherry, S; Dybing, J K; Davis, N L et al. (2000) Influenza virus (A/HK/156/97) hemagglutinin expressed by an alphavirus replicon system protects chickens against lethal infection with Hong Kong-origin H5N1 viruses. Virology 278:55-9
Bernard, K A; Klimstra, W B; Johnston, R E (2000) Mutations in the E2 glycoprotein of Venezuelan equine encephalitis virus confer heparan sulfate interaction, low morbidity, and rapid clearance from blood of mice. Virology 276:93-103
Davis, N L; Caley, I J; Brown, K W et al. (2000) Vaccination of macaques against pathogenic simian immunodeficiency virus with Venezuelan equine encephalitis virus replicon particles. J Virol 74:371-8
Aronson, J F; Grieder, F B; Davis, N L et al. (2000) A single-site mutant and revertants arising in vivo define early steps in the pathogenesis of Venezuelan equine encephalitis virus. Virology 270:111-23
MacDonald, G H; Johnston, R E (2000) Role of dendritic cell targeting in Venezuelan equine encephalitis virus pathogenesis. J Virol 74:914-22
Charles, P C; Brown, K W; Davis, N L et al. (1997) Mucosal immunity induced by parenteral immunization with a live attenuated Venezuelan equine encephalitis virus vaccine candidate. Virology 228:153-60
Davis, N L; Brown, K W; Johnston, R E (1996) A viral vaccine vector that expresses foreign genes in lymph nodes and protects against mucosal challenge. J Virol 70:3781-7

Showing the most recent 10 out of 27 publications