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Journal of Virology, August 2001, p. 7420-7428, Vol. 75, No. 16
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.16.7420-7428.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

High Numbers of Viral RNA Copies in the Central Nervous System of Mice during Persistent Infection with Theiler's Virus

Mark Trottier,1,2 Pat Kallio,1 Wei Wang,1,3 and Howard L. Lipton1,2,4,5,*

Department of Neurology, Evanston Hospital,1 and Departments of Neurology,5 Microbiology-Immunology,4 and Biochemistry, Molecular Biology and Cell Biology,2 and Neuroscience Graduate Program,3 Northwestern University, Evanston, Illinois

Received 16 January 2001/Accepted 16 May 2001

The low-neurovirulence Theiler's murine encephalomyelitis viruses (TMEV), such as BeAn virus, cause a persistent infection of the central nervous system (CNS) in susceptible mouse strains that results in inflammatory demyelination. The ability of TMEV to persist in the mouse CNS has traditionally been demonstrated by recovering infectious virus from the spinal cord. Results of infectivity assays led to the notion that TMEV persists at low levels. In the present study, we analyzed the copy number of TMEV genomes, plus- to minus-strand ratios, and full-length species in the spinal cords of infected mice and infected tissue culture cells by using Northern hybridization. Considering the low levels of infectious virus in the spinal cord, a surprisingly large number of viral genomes (mean of 3.0 × 109) was detected in persistently infected mice. In the transition from the acute (approximately postinfection [p.i.] day 7) to the persistent (beginning on p.i. day 28) phase of infection, viral RNA copy numbers steadily increased, indicating that TMEV persistence involves active viral RNA replication. Further, BeAn viral genomes were full-length in size; i.e., no subgenomic species were detected and the ratio of BeAn virus plus- to minus-strand RNA indicated that viral RNA replication is unperturbed in the mouse spinal cord. Analysis of cultured macrophages and oligodendrocytes suggests that either of these cell types can potentially synthesize high numbers of viral RNA copies if infected in the spinal cord and therefore account for the heavy viral load. A scheme is presented for the direct isolation of both cell types directly from infected spinal cords for further viral analyses.

* Corresponding author. Mailing address: Department of Neurology, Multiple Sclerosis Research Center, Evanston Hospital, Northwestern University, 2650 Ridge Ave., Evanston, IL 60201-1782. Phone: (847) 570-2168. Fax: (847) 570-1568. E-mail: hllipton{at}merle.acns.nwu.edu.

Journal of Virology, August 2001, p. 7420-7428, Vol. 75, No. 16
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.16.7420-7428.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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