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Journal of Virology, April 2006, p. 3644-3649, Vol. 80, No. 7
0022-538X/06/$08.00+0 doi:10.1128/JVI.80.7.3644-3649.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.
Infection and Persistence of Rhesus Monkey Rhadinovirus in Immortalized B-Cell Lines
John P. Bilello,1 Sabine M. Lang,1,
Fred Wang,2
Jon C. Aster,3 and
Ronald C. Desrosiers1*
New England Primate Research Center, Harvard Medical School, Southborough, Massachusetts 01772,1 Channing Laboratory,2 Pathology Department, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 021153
Received 7 July 2005/ Accepted 18 January 2006
Similar to its close relative human herpesvirus 8, rhesus monkey rhadinovirus (RRV) persists predominantly in B cells of its natural host. Rhesus monkey B-cell lines immortalized by the Epstein-Barr-related virus from rhesus monkeys (rhEBV) were used as targets for infection by RRV. These cultured B cells were susceptible to infection by RRV and continued to produce low titers of RRV for months of continuous culture. Infection by RRV did not detectably alter the growth rates of these B-cell lines when it was measured at standard or reduced serum concentrations. Depending on the cell line, 5 to 40% of the B cells stained positive for the RRV genome by fluorescence in situ hybridization (FISH). Most RRV-positive cells showed a fine punctate nuclear staining pattern consistent with latent infection, while a small minority of cells (0.2 to 1%) contained large, intensely staining nuclear foci consistent with productive, replicative infection. Greater than 90% of the cells were rhEBV genome positive in a pattern consistent with latent infection, and again only a small minority of cells showed a productive, replicative staining pattern. Dual, two-color FISH staining revealed coinfection of numerous cells with both RRV and rhEBV, but productive replication of RRV and rhEBV was always observed in separate cells, never in the same cell. Thus, productive replication of RRV is unlinked to that of rhEBV; factors that influence activation to productive replication act separately on RRV and rhEBV, even within the same cell. The percentage of B cells expressing green fluorescent protein (GFP) early after infection with a recombinant RRV containing a GFP reporter gene was dose dependent and at a low multiplicity of infection increased progressively over time until 14 to 17 days after infection. These results establish a naturalistic cell culture system for the study of infection and persistence by RRV in rhesus monkey B cells.
* Corresponding author. Mailing address: New England Primate Research Center, Harvard Medical School, One Pine Hill Drive, Box 9102, Southborough, MA 01772-9102. Phone: (508) 624-8160. Fax: (508) 624-8190. E-mail: ronald_desrosiers{at}hms.harvard.edu.
Present address: Department of Pathology, Yale University Medical School, New Haven, CT 06510.
Journal of Virology, April 2006, p. 3644-3649, Vol. 80, No. 7
0022-538X/06/$08.00+0 doi:10.1128/JVI.80.7.3644-3649.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.
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