The Use of Chimeric Venezuelan Equine Encephalitis Viruses as an Approach for the Molecular Identification of Natural Virulence Determinants

doi:10.1128/JVI.74.9.4258-4263.2000

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Journal of Virology, May 2000, p. 4258-4263, Vol. 74, No. 9
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

The Use of Chimeric Venezuelan Equine Encephalitis Viruses as an Approach for the Molecular Identification of Natural Virulence Determinants

Ann M. Powers,¹^,^* Aaron C. Brault,¹ Richard M. Kinney,² and Scott C. Weaver¹

Center for Tropical Diseases and Department of Pathology, University of Texas Medical Branch, Galveston, Texas 77555-0609,¹ and Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Services, Fort Collins, Colorado 80522²

Received 6 October 1999/Accepted 31 January 2000

Venezuelan equine encephalitis (VEE) virus antigenic subtypes and varieties are considered either epidemic/epizootic or enzootic.In addition to epidemiological differences between the epidemicand enzootic viruses, several in vitro and in vivo laboratorymarkers distinguishing the viruses have been identified, includingdifferential plaque size, sensitivity to interferon (IFN), andvirulence for guinea pigs. These observations have been shownto be useful predictors of natural, equine virulence and epizooticpotential. Chimeric viruses containing variety IAB (epizootic)nonstructural genes with variety IE (enzootic) structural genes(VE/IAB-IE) or IE nonstructural genes and IAB structural genes(IE/IAB) were constructed to systematically analyze and map viralphenotype and virulence determinants. Plaque size analysis showedthat both chimeric viruses produced a mean plaque diameter thatwas intermediate between those of the parental strains. Additionally,both chimeric viruses showed intermediate levels of virus replicationand virulence for guinea pigs compared to the parental strains.However, IE/IAB produced a slightly higher viremia and an averagesurvival time 2 days shorter than the VE/IAB-IE virus. Finally,IFN sensitivity assays revealed that only one chimera, VE/IAB-IE,was intermediate between the two parental types. The second chimera,containing the IE nonstructural genes, was at least five timesmore sensitive to IFN than the IE parental virus and greater than50 times more sensitive than the IAB parent. These results implicateviral components in both the structural and nonstructural portionsof the genome in contributing to the epizootic phenotype and indicatethe potential for epidemic emergence from the IE enzootic VEEviruses.

^* Corresponding author. Mailing address: Department of Pathology & Center for Tropical Diseases, University of Texas MedicalBranch, 301 University Blvd., Galveston, TX 77555-0609. Phone:(409) 747-2440. Fax: (409) 747-2415. E-mail: ampowers{at}utmb.edu.

Journal of Virology, May 2000, p. 4258-4263, Vol. 74, No. 9
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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