The Kaposi's sarcoma-associated herpesvirus G protein-coupled receptor has broad signaling effects in primary effusion lymphoma cells
The Kaposi's sarcoma-associated herpesvirus G protein-coupled receptor has broad signaling effects in primary effusion lymphoma cells
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DOI：
10.1128/jvi.77.1.57-67.2003
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发表时间：
2003-01-01
影响因子：
5.4
通讯作者：
Cesarman, E
中科院分区：
文献类型：
--
作者：
Cannon, M;Philpott, NJ;Cesarman, E
卡波西肉瘤相关疱疹病毒（KSHV/人疱疹病毒8 [HHV-8]）是一种导致卡波西肉瘤和原发性积液性淋巴瘤（PEL）的γ -2疱疹病毒。KSHV是一种嗜淋巴病毒，它窃取了许多与炎症、细胞周期控制和血管生成有关的哺乳动物基因。其中包括早期裂解病毒G蛋白偶联受体（vGPCR），它是人类白细胞介素-8 （IL-8）受体的同源物。当表达时，vGPCR具有组成性活性，可以通过丝裂原和应激激活激酶发出信号。在某些模型中，它激活NF-kappaB和激活蛋白1 （AP-1）的转录潜能，诱导血管内皮生长因子（VEGF）的产生。尽管vGPCR对所有kshv介导的疾病的发病机制很重要，但对造血细胞中的vGPCR活性知之甚少。为了研究vGPCR在这些细胞中的信号传导潜能和下游作用，我们开发了在诱导启动子控制下表达vGPCR的PEL细胞系。将四环素介导诱导所需的序列克隆到含有腺相关病毒2型元件的质粒中，以提高整合效率。这种新型质粒允许在自然感染的kshv阳性淋巴细胞中研究vGPCR活性。我们发现vGPCR可以激活PEL细胞中的ERK-2和p38。此外，它在AP-1、NF-kappaB、CREB和NFAT的控制下增加了报告基因的转录，NFAT是一种Ca2+依赖性转录因子，对KSHV裂解基因的表达很重要。vGPCR还增加了KSHV开放阅读框50和57的转录，从而显示出影响病毒转录模式的广泛潜力。最后，vGPCR信号导致KSHV- 6和VEGF的PEL细胞细化增加，这两种生长因子参与KSHV介导的疾病发病机制。
Kaposi's sarcoma-associated herpesvirus (KSHV/human herpesvirus 8 [HHV-8]) is a gamma-2-herpesvirus responsible for Kaposi's sarcoma as well as primary effusion lymphoma (PEL). KSHV is a lymphotropic virus that has pirated many mammalian genes involved in inflammation, cell cycle control, and angiogenesis. Among these is the early lytic viral G protein-coupled receptor (vGPCR), a homologue of the human interleukin-8 (IL-8) receptor. When expressed, vGPCR is constitutively active and can signal via mitogen- and stress-activated kinases. In certain models it activates the transcriptional potential of NF-kappaB and activator protein 1 (AP-1) and induces vascular endothelial growth factor (VEGF) production. Despite its importance to the pathogenesis of all KSHV-mediated disease, little is known about vGPCR activity in hematopoietic cells. To study the signaling Potential and downstream effects of vGPCR in such cells, we have developed PEL cell lines that express vGPCR under the control of an inducible promoter. The sequences required for tetracycline-mediated induction were cloned into a plasmid containing adeno-associated virus type 2 elements to enhance integration efficiency. This novel plasmid permitted studies of vGPCR activity in naturally infected KSHV-positive lymphocytes. We show that vGPCR activates ERK-2 and p38 in PEL cells. In addition, it increases the transcription of reporter genes under the control of AP-1, NF-kappaB, CREB, and NFAT, a Ca2+-dependent transcription factor important to KSHV lytic gene expression. vGPCR also increases the transcription of KSHV open reading frames 50 and 57, thereby displaying broad potential to affect viral transcription patterns. Finally, vGPCR signaling results in increased PEL cell elaboration of KSHV vIL-6 and VEGF, two growth factors involved in KSHV-mediated disease pathogenesis.