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 infe ctious Kaposi's sarcoma herpesvirus (KSHV) supernatants was established fro m the ascitic fluid of a human immunodeficiency virus-positive patient. Flo w cytometry showed strong expression of CD45 and lambda light-chain restric tion, Southern blot hybridization showed immunoglobulin heavy-chain gene re arrangements in the tumor and the resultant cell line consistent with B-cel l lineage. Expression of viral genes was assessed by reverse transcription- PCR and immunohistochemistry. Only latent Epstein-Barr virus (EBV) gene exp ression was detected, and this was at a low level. In contrast, lytic and l atent 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 h uman dermal microvascular endothelial cells (DMVECs) with KSHV than superna tants from BC-3 or BCP-1 PEL cell lines. Quantitation of viral yields produ ced by the PEL lines showed at least 2 orders of magnitude more DNase I-res istant KSHV DNA in the JSC-1 supernatant compared to BC-3 or BCP-1 supernat ants. KSHV infection in DMVECs was associated with a change from a cobblest one to a spindle shape, LANA expression, and an increased number of mitoses .