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一种新的原发性渗出性淋巴瘤衍生细胞系产生了含有高传染性卡波西肉瘤疱疹病毒的上清液。

A new primary effusion lymphoma-derived cell line yields a highly infectious Kaposi's sarcoma herpesvirus-containing supernatant.

作者信息

Cannon J S, Ciufo D, Hawkins A L, Griffin C A, Borowitz M J, Hayward G S, Ambinder R F

机构信息

Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.

出版信息

J Virol. 2000 Nov;74(21):10187-93. doi: 10.1128/jvi.74.21.10187-10193.2000.

Abstract

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.

摘要

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。

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