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Adaptation of Coronavirus JHM to Persistent Infection of Murine Sac(-) Cells

Herbert N. Baybutt

Michael J. Carter

Institut für Virologie und Immunbiologie der Universität Würzburg, Versbacher Strasse 7, D-8700 Würzburg, Federal Republic of Germany

Coronaviruses can establish persistent infections in the central nervous system of rodents, and these are associated with demyelinating encephalomyelitis. The effects of persistence on the virus are difficult to study in vivo but may have a crucial influence on the course of infection. We therefore produced a persistent infection in vitro using the neurotropic coronavirus JHM, in order to investigate the events underlying the establishment of such an infection and the adaptation of the virus to persistence. The persistent infection was maintained for over 115 passages and continued to release high levels of infectious virus. During the 18 months of culture the number of cells expressing virus antigen detected by indirect immune fluorescence decreased to 40%. Analysis showed that the carried virus contained a significant proportion of heterogeneous temperature-sensitive mutants. All virus clones isolated possessed the capacity to induce a more productive growth cycle, a less pronounced cytopathic effect and showed a much reduced neurovirulence when inoculated into newborn and weanling rats. Evidence for structural changes involving the surface peplomer protein (E2) was obtained using hybridoma antibodies, which neutralized the parental JHM virus but not the JHM-Pi virus. Defective interfering particles and interferon activities have been excluded as possible agents instrumental in the establishment and maintenance of the chronic infection, and we suggest that the emergence of virus variants of lowered virulence is central to these processes.

Keywords: coronavirus JHM, persistent infection, adaptation, virulence

Present address: Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.

Present address: Department of Virology, Royal Victoria Infirmary, Newcastle-upon-Tyne, U.K.

Received 24 November 1983; accepted 1 February 1984.

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