Generation of a doxycycline-inducible KSHV producer cell line of endothelial origin: maintenance of tight latency with efficient reactivation upon induction.
Generation of a doxycycline-inducible KSHV producer cell line of endothelial origin: maintenance of tight latency with efficient reactivation upon induction.
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DOI：
10.1016/j.jviromet.2011.03.012
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发表时间：
2011-06
影响因子：
3.1
通讯作者：
Ganem D
中科院分区：
文献类型：
--
作者：
Myoung J;Ganem D
卡波西肉瘤相关疱疹病毒（KSHV）是卡波西肉瘤（KS）以及至少两种B细胞淋巴增生性疾病的病原体：原发性渗出性淋巴瘤（PEL）和多中心型卡斯特曼病（MCD）。源自PEL的B细胞处于潜伏感染状态，可用诸如佛波醇12-十四酸酯13-乙酸酯（TPA）或丁酸盐等化学试剂处理，诱导其进入裂解性复制，这些化学试剂对宿主细胞信号传导和染色质结构具有多效性作用。大多数此类细胞系还表现出中等程度的自发裂解诱导，这使得潜伏期分析变得复杂。在此，我们描述了源自SLK内皮细胞的潜伏感染细胞系的构建，这些细胞系具有以下特点：（i）对KSHV潜伏期具有严格控制，几乎无自发再激活现象；（ii）可通过强力霉素高效诱导，无需使用具有多效性的诱导剂。这些细胞可产生大量有感染性的KSHV，对于研究潜伏 - 裂解转换以及裂解性复制对宿主细胞生物学的影响应具有实用价值。
Kaposi’s sarcoma-associated herpes virus (KSHV) is the etiological agent of Kaposi’s sarcoma (KS) and at least two B cell lymphoproliferative diseases: primary effusion lymphoma (PEL) and multicentric Castleman’s disease (MCD). B cells derived from PEL are latently infected, and can be induced to lytic replication by treatment with chemical agents like TPA or butyrate, which have pleiotropic effects on host cell signaling and chromatin structure. Most of these lines also display moderate levels of spontaneous lytic induction, which complicates analysis of latency. Here we describe the creation of latently-infected cell lines derived from SLK endothelial cells that (i) display tight control of KSHV latency, with little spontaneous reactivation and (ii) are efficiently inducible by doxycycline, avoiding the need for pleiotropic inducing agents. These cells produce substantial quantities of infectious KSHV, and should be useful for studies of the latent-lytic switch and the impact of lytic replication on host cell biology.