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Journal of Virology, June 2001, p. 4955-4963, Vol. 75, No. 11
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.11.4955-4963.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Kinetics of Murine Gammaherpesvirus 68 Gene Expression following Infection of Murine Cells in Culture and in Mice

Rosemary Rochford,1,* Mary L. Lutzke,1 Rosiane S. Alfinito,1 Anaira Clavo,1,dagger and Rhonda D. Cardin2

Department of Epidemiology, University of Michigan, Ann Arbor, Michigan 48109,1 and Pfizer Global Research and Development, Ann Arbor, Michigan 481052

Received 29 November 2000/Accepted 2 March 2001

A model system to study the pathogenesis of gammaherpesvirus infections is the infection of mice with murine gammaherpesvirus 68 (MHV-68). To define the kinetics of infection, we developed an RNase protection assay to quantitate gene expression from lytic (K3, Rta, M8, DNA polymerase [DNA pol], and gB) and candidate latency (M2, M3, M9, M11, ORF73, and ORF74) genes. All candidate latency genes were expressed during lytic infection of 3T3 cells. Four kinetic classes of transcripts were observed following infection of 3T3 cells: immediate-early (K3, Rta, M8, and ORF73), early (DNA pol), early-late (M3, M11, and ORF74), and late (M2, M9, and gB). To assess the kinetics of viral gene expression in vivo, lungs, spleens, and mediastinal lymph nodes (MLN) were harvested from MHV-68-infected mice. All transcripts were expressed between 3 and 6 days postinfection (dpi) in the lungs. In the spleen, K3, M3, M8, and M9 transcripts were expressed between 10 and 16 dpi when latency is established. The K3, M3, M8, M9, and M11 transcripts were detected in the MLN from 2 through 16 dpi. This is the first demonstration of MHV-68 gene expression in the MLN. Importantly, our data showed that MHV-68 has different kinetics of gene expression at different sites of infection. Furthermore, we demonstrated that K3, a gene recently shown to encode a protein that downregulates major histocompatibility complex class I on the surface of cells, is expressed during latency, which argues for a role of K3 in immune evasion during latent infection.

* Corresponding author. Mailing address: Department of Epidemiology, University of Michigan, 109 Observatory Rd., Ann Arbor, MI 48109-2029. Phone: (734) 764-5469. Fax: (734) 764-3192. E-mail: rochford{at}umich.edu.

dagger Present address: Center for Neurovirology and Cancer Biology, Temple University, Philadelphia, PA 19122.

Journal of Virology, June 2001, p. 4955-4963, Vol. 75, No. 11
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.11.4955-4963.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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