Naranatt Pramod P, Krishnan Harinivas H, Svojanovsky Stan R, Bloomer Clark, Mathur Sachin, Chandran Bala
Department of Microbiology, Molecular Genetics and Immunology, Bioinformatics Core, and Microarray Core, The University of Kansas Medical Center, Kansas City, Kansas 66160, USA.
Cancer Res. 2004 Jan 1;64(1):72-84. doi: 10.1158/0008-5472.can-03-2767.
Kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8) is etiologically linked to the endothelial tumor Kaposi's sarcoma and with two lymphoproliferatve disorders, primary effusion lymphoma and multicentric Castleman's disease. HHV-8 infects a variety of target cells both in vivo and in vitro, binds to the in vitro target cells via cell surface heparan sulfate, and uses the alpha(3)beta(1) integrin as one of the entry receptors. Within minutes of infection, HHV-8 induced the integrin-mediated signaling pathways and morphological changes in the target cells (S. M. Akula et al., Cell, 108: 407-419, 2002; P. P. Naranatt et al., J. Virol., 77: 1524-1539, 2003). As an initial step toward understanding the role of host genes in HHV-8 infection and pathogenesis, modulation of host cell gene expression immediately after infection was examined. To reflect HHV-8's broad cellular tropism, mRNAs collected at 2 and 4 h after infection of primary human endothelial [human adult dermal microvascular endothelial cells (HMVECd)] and foreskin fibroblast [human foreskin fibroblast (HFF)] cells and human B cell line (BJAB) were analyzed by oligonucleotide array with approximately 22,000 human transcripts. With a criteria of >2-fold gene induction as significant, approximately 1.72% of the genes were differentially expressed, of which, 154 genes were shared by at least two cells and 33 genes shared by all three cells. HHV-8-induced transcriptional profiles in the endothelial and fibroblast cells were closely similar, with substantial differences in the B cells. In contrast to the antiapoptotic regulators induced in HMVECd and HFF cells, proapoptotic regulators were induced in the B cells. A robust increase in the expression of IFN-induced genes suggestive of innate immune response induction was observed in HMVECd and HFF cells, whereas there was a total lack of immunity related protein inductions in B cells. These striking cell type-specific behaviors suggest that HHV-8-induced host cell gene modulation events in B cells may be different compared with the adherent endothelial and fibroblast target cells. Functional clustering of modulated genes identified several host molecules hitherto unknown to HHV-8 infection. These results indicate that early during infection, HHV-8 reprograms the host transcriptional machinery regulating a variety of cellular processes including apoptosis, transcription, cell cycle regulation, signaling, inflammatory response, and angiogenesis, all of which may play important roles in the biology and pathogenesis of HHV-8.
卡波西肉瘤相关疱疹病毒（KSHV/HHV - 8）在病因上与内皮肿瘤卡波西肉瘤以及两种淋巴增殖性疾病，即原发性渗出性淋巴瘤和多中心性Castleman病相关。HHV - 8在体内和体外均可感染多种靶细胞，通过细胞表面硫酸乙酰肝素与体外靶细胞结合，并将α(3)β(1)整合素作为一种进入受体。在感染后的数分钟内，HHV - 8诱导靶细胞中的整合素介导的信号通路和形态变化（S.M.阿库拉等人，《细胞》，第108卷：407 - 419页，2002年；P.P.纳拉纳特等人，《病毒学杂志》，第77卷：1524 - 1539页，2003年）。作为理解宿主基因在HHV - 8感染和发病机制中作用的第一步，研究了感染后立即对宿主细胞基因表达的调节。为反映HHV - 8广泛的细胞嗜性，对感染原代人内皮细胞[成人真皮微血管内皮细胞（HMVECd）]、包皮成纤维细胞[人包皮成纤维细胞（HFF）]和人B细胞系（BJAB）2小时和4小时后收集的mRNA，用约22,000个人类转录本的寡核苷酸阵列进行分析。以基因诱导>2倍为显著标准，约1.72%的基因差异表达，其中，至少两个细胞共有的基因有154个，三个细胞共有的基因有33个。HHV - 8在内皮细胞和成纤维细胞中诱导的转录谱非常相似，而在B细胞中有显著差异。与在HMVECd和HFF细胞中诱导的抗凋亡调节因子相反，B细胞中诱导的是促凋亡调节因子。在HMVECd和HFF细胞中观察到IFN诱导基因表达的强劲增加，提示先天免疫反应被诱导，而B细胞中完全缺乏免疫相关蛋白的诱导。这些显著的细胞类型特异性行为表明，与贴壁的内皮细胞和成纤维细胞靶细胞相比，HHV - 8在B细胞中诱导的宿主细胞基因调节事件可能不同。对调节基因的功能聚类鉴定出了一些迄今未知与HHV - 8感染相关的宿主分子。这些结果表明，在感染早期，HHV - 8对宿主转录机制进行重新编程，调节包括凋亡、转录、细胞周期调控、信号传导、炎症反应和血管生成在内的多种细胞过程，所有这些过程可能在HHV - 8的生物学和发病机制中发挥重要作用。
