Characterization of Kaposi sarcoma-associated herpesvirus/human herpesvirus-8 infection of human vascular endothelial cells: early events
Characterization of Kaposi sarcoma-associated herpesvirus/human herpesvirus-8 infection of human vascular endothelial cells: early events
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DOI：
10.1182/blood.v100.3.888
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发表时间：
2002-08-01
期刊：
影响因子：
20.3
通讯作者：
Fingeroth, JD
中科院分区：
文献类型：
--
作者：
Dezube, BJ;Zambela, M;Fingeroth, JD
卡波西肉瘤相关疱疹病毒（KSHV）/人疱疹病毒-8（HHV-8）与卡波西肉瘤（KS）有因果关系。然而，缺乏一种细胞培养系统，有效地再现复合机制的启动和维持KS内皮细胞中的HHV-8感染（裂解和潜伏），留下了重要的问题没有答案。在这里，我们报告了一种培养系统，其中伴随HHV-8感染的最早事件可以在原代内皮细胞中进行调查。将HHV-8与微血管真皮内皮细胞（MVDEC）的结合直接与HHV-8相关疾病中涉及的其他主要靶细胞进行比较。通过电子显微镜监测病毒在MVDEC内的附着、融合、内化和运输。对基因组构型的研究表明，病毒DNA在进入时发生快速环化，尽管在感染后72小时，线性DNA积累，并且可以检测到早期和晚期裂解RNA（T1.1，K8.1）。在第8天首次检测到潜伏期转录本（LT 1/LT 2），表明裂解和潜伏感染均启动。虽然大多数裂解转录累积，直到通过，开放阅读框-74 RNA波动与一个固定的周期性，这表明早期复制感染后的MVDEC是同步的。(C)2002年，美国血液学会。
Kaposi sarcoma-associated herpesvirus (KSHV)/human herpesvirus-8 (HHV-8) is causally associated with Kaposi sarcoma (KS). The absence of a cell culture system that effectively reproduces the composite mechanisms governing initiation and maintenance of HHV-8 infection (lytic and latent) in KS endothelial cells, however, has left important questions unanswered. Here, we report a culture system in which the earliest events that accompany HHV-8 infection could be surveyed in primary endothelial cells. Binding of HHV-8 to microvascular dermal endothelial cells (MVDECs) was directly compared with other primary target cells implicated in HHV-8-associated diseases. Virus attachment, fusion, internalization and transport within MVDECs was monitored by electron microscopy. Studies of genome configuration revealed that rapid circularization of the viral DNA occurred on entry, though by 72 hours after infection linear DNAs accumulated and early as well as late lytic RNAs (T1.1, K8.1) could be detected. The latency transcripts (LT1/LT2) were first detected on day 8, demonstrating that both lytic and latent infection were initiated. Although most lytic transcripts accrued until passage, open-reading frame-74 RNAs fluctuated with a fixed periodicity, suggesting that early replication after infection of MVDECs was synchronous. (C) 2002 by The American Society of Hematology.