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Journal of Virology, June 1999, p. 5149-5155, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Characterization of Monoclonal Antibodies Raised against the Latent Nuclear Antigen of Human Herpesvirus 8

Paul Kellam,^1,* Dimitra Bourboulia,¹ Nicolas Dupin,¹ Chris Shotton,¹ Cyril Fisher,² Simon Talbot,^1, Chris Boshoff,^1,3 and Robin A. Weiss¹

Institute of Cancer Research, Chester Beatty Laboratories, London SW3 6JB,¹ Royal Marsden Hospital, London SW3 6JJ,² and Department of Oncology, Royal Free and University College Medical School, University College London, London W1P 6BT,³ United Kingdom

Received 8 October 1998/Accepted 16 February 1999

Human herpesvirus 8 (HHV-8; also designated Kaposi's sarcoma-associated herpesvirus) is the likely etiological agent of Kaposi's sarcoma (KS). HHV-8 encodes a latent nuclear antigen (LNA) which is the product of the viral gene orf 73. LNA is recognized by most infected patient sera and is the basis of current immunofluorescence assays used in epidemiological studies of HHV-8 infection. Here we describe the characterization of four monoclonal antibodies raised to the C-terminal third of LNA-glutathione S-transferase fusion proteins. These monoclonal antibodies recognized discrete linear epitopes within the C terminus and repetitive region of LNA, detected antigen in primary effusion lymphoma (PEL) cells, and precipitated a 220- to 230-kDa protein doublet corresponding to LNA from HHV-8-infected PEL cell lines. In situ immunocytochemistry of KS lesions with these antibodies show that LNA is extensively expressed in KS spindle cells.

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^* Corresponding author. Present address: Windeyer Institute of Medical Sciences, University College London, 46 Cleveland St., London W1P 6DB, United Kingdom. Phone: 44-171-504-9343. Fax: 44-171-387-3310. E-mail: p.Kellam{at}ucl.ac.uk.

Present address: Department of Medical Microbiology, Edinburgh University Medical School, Edinburgh EH8 9AG, United Kingdom.

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Journal of Virology, June 1999, p. 5149-5155, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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