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Journal of Virology, August 2006, p. 7873-7884, Vol. 80, No. 16
0022-538X/06/$08.00+0     doi:10.1128/JVI.00529-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Dissemination of Bovine Leukemia Virus-Infected Cells from a Newly Infected Sheep Lymph Node

B. E. Fulton Jr.,^1,2, M. Portella,^2, and K. Radke^1,2*

Department of Animal Science,² Graduate Group in Microbiology, University of California, Davis, California 95616-8521¹

Received 14 March 2006/ Accepted 3 June 2006

To investigate the early establishment of bovine leukemia virus (BLV) infection, we injected BLV-infected or mock-infected allogeneic cells into the shoulder of sheep in which an efferent lymphatic duct of the draining prescapular lymph node had been cannulated. Rare mononuclear cells acting as centers of BLV infection in culture were present within 4 to 6 days in efferent lymph and within 6 to 10 days in blood. Soon after BLV injection, immunoglobulin M⁺ (IgM⁺) and CD8⁺ cells increased in efferent lymph and oscillated reciprocally in frequency. CD8⁺ blasts increased on days 4 to 6, when infectious centers increased 100-fold in lymph. On days 6 and 7, both lymph and blood were enriched with CD8⁺ cells that were labeled late on day 5 with an intravenous pulse of 5-bromo-2'-deoxyuridine (BrdU). Lymph, but not blood, was enriched with BrdU⁺ B cells on day 7. Capsid-specific antibodies became detectable in efferent lymph on days 6 to 8 and surface glycoprotein-specific antibodies on day 9, preceding their detection in serum by 9 to 14 days. Systemic dissemination of BLV-infected cells was thus accompanied by an increase in proliferating CD8⁺ cells and the onset of BLV-specific antibodies in lymph. Infectious centers reached maximum frequencies of 0.2% in lymph by days 11 to 13, and then their frequencies increased by 5- to 40-fold in blood cells, suggesting that many infected blood cells do not recirculate back into lymph. Beginning on days 10 to 13, a subpopulation of B cells having high levels of surface IgM increased sharply in peripheral blood. Such cells were not present in lymph. After a day 16 pulse of BrdU, recently proliferated cells that stained intensely for surface IgM appeared in blood within 15 h. Predominantly B lymphocytes contained the viral capsid protein when lymph and blood cells were cultured briefly to allow BLV expression. However, both early in lymph and later in blood, BrdU⁺ B cells greatly exceeded productively infected cells, indicating that new BLV infections stimulate proliferation of two different populations of B cells.

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* Corresponding author. Mailing address: Department of Animal Science, University of California, One Shields Ave., Davis, CA 95616-8521. Phone: (530) 752-9025. Fax: (530) 752-0175. E-mail: KLRadke{at}ucdavis.edu.

Present address: Intrexon Corporation, Blacksburg, VA 24060.

Present address: BioPlex Division, Bio-Rad Laboratories, Benicia, CA 94510.

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Journal of Virology, August 2006, p. 7873-7884, Vol. 80, No. 16
0022-538X/06/$08.00+0     doi:10.1128/JVI.00529-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.