Cannon J S, Ciufo D, Hawkins A L, Griffin C A, Borowitz M J, Hayward G S, Ambinder R F
Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
J Virol. 2000 Nov;74(21):10187-93. doi: 10.1128/jvi.74.21.10187-10193.2000.
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.
一株原发性渗出性淋巴瘤(PEL)细胞系JSC-1,它能产生具有高度传染性的卡波西肉瘤疱疹病毒(KSHV)上清液,该细胞系是从一名人类免疫缺陷病毒阳性患者的腹水中建立的。流式细胞术显示CD45呈强表达且存在λ轻链限制性。Southern印迹杂交显示肿瘤及由此产生的细胞系中免疫球蛋白重链基因重排,与B细胞谱系一致。通过逆转录PCR和免疫组织化学评估病毒基因的表达。仅检测到潜伏性EB病毒(EBV)基因表达,且水平较低。相比之下,检测到了KSHV的裂解性和潜伏性基因表达。十四酰佛波醇乙酸酯和丁酸盐上调了KSHV的裂解性表达,但未上调EBV的裂解性表达。与BC-3或BCP-1 PEL细胞系的上清液相比,JSC-1的病毒上清液在用KSHV感染原代人真皮微血管内皮细胞(DMVECs)方面效率更高。对PEL细胞系产生的病毒产量进行定量分析显示,与BC-3或BCP-1的上清液相比,JSC-1上清液中对脱氧核糖核酸酶I有抗性的KSHV DNA至少多2个数量级。DMVECs中的KSHV感染与细胞从鹅卵石样形态转变为纺锤形、LANA表达以及有丝分裂数量增加有关。