A new primary effusion lymphoma-derived cell line yields a highly infectious Kaposi's sarcoma herpesvirus-containing supernatant.
A primary effusion lymphoma (PEL) cell line, JSC-1, that yields highly infectious Kaposi's sarcoma herpesvirus (KSHV) supernatants was established from the ascitic fluid of a human immunodeficiency virus-positive patient. Flow cytometry showed strong expression of CD45 and lambda light-chain restriction. Southern blot hybridization showed immunoglobulin heavy-chain gene rearrangements in the tumor and the resultant cell line consistent with B-cell lineage. Expression of viral genes was assessed by reverse transcription-PCR and immunohistochemistry. Only latent Epstein-Barr virus (EBV) gene expression was detected, and this was at a low level. In contrast, lytic and latent KSHV gene expression were detected. Tetradecanoyl phorbol acetate and butyrate upregulated KSHV lytic expression, but not EBV lytic expression. Viral supernatant from JSC-1 was much more efficient at infecting primary human dermal microvascular endothelial cells (DMVECs) with KSHV than supernatants from BC-3 or BCP-1 PEL cell lines. Quantitation of viral yields produced by the PEL lines showed at least 2 orders of magnitude more DNase I-resistant KSHV DNA in the JSC-1 supernatant compared to BC-3 or BCP-1 supernatants. KSHV infection in DMVECs was associated with a change from a cobblestone to a spindle shape, LANA expression, and an increased number of mitoses.
Cannon
J S
JS
Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Ciufo
D
D
Hawkins
A L
AL
Griffin
C A
CA
Borowitz
M J
MJ
Hayward
G S
GS
Ambinder
R F
RF
D001202:Ascitic Fluid / Q000166:cytology* / Q000821:virology; D004730:Endothelium, Vascular / Q000166:cytology / Q000821:virology; D005455:Fluorescent Antibody Technique; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D016483:Lymphoma, AIDS-Related / Q000821:virology*; D008297:Male; D008875:Middle Aged; D012867:Skin / Q000098:blood supply; D014407:Tumor Cells, Cultured; D014771:Virion / Q000502:physiology