Long-term-infected telomerase-immortalized endothelial cells: a model for Kaposi's sarcoma-associated herpesvirus latency in vitro and in vivo
Long-term-infected telomerase-immortalized endothelial cells: a model for Kaposi's sarcoma-associated herpesvirus latency in vitro and in vivo
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DOI：
10.1128/jvi.80.10.4833-4846.2006
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发表时间：
2006-05-01
影响因子：
5.4
通讯作者：
Renne, Rolf
中科院分区：
文献类型：
--
作者：
An, Feng-Qi;Folarin, Hope Merlene;Renne, Rolf
卡波西肉瘤相关疱疹病毒(KSHV)与卡波西肉瘤(KS)、原发渗出性淋巴瘤(PEL)和多中心性Castleman病有关。大多数KS肿瘤细胞潜伏感染KSHV，并且是内皮源性的。虽然PEL来源的细胞系无限期地保持KSHV，但到目前为止，所有KS肿瘤来源的细胞在体外培养时都失去了病毒基因组。为了研究KSHV潜伏期和内皮细胞中的肿瘤发生，我们产生了端粒酶永生化的人脐静脉内皮细胞。感染后48h，活性细胞表达所有KSHV潜伏基因，RTA/Orf50可诱导有效的裂解复制。与以前的模型类似，受感染的培养物逐渐失去了病毒的异构体。然而。我们还首次获得了两个内皮细胞系，在没有选择的情况下，KSHV的内切体可以无限期地保持。KSHV的长期维持与RTA-Orf50反应的重新激活丧失和完全致癌转化相关。长期感染细胞(LTC)在软琼脂中生长，在无血清条件下增殖。在裸鼠体内形成肿瘤的是LTC，而不是亲代活性细胞。这些肿瘤表达高水平的潜伏期相关核抗原(LANA)，并表达KS(Lyve-1)中发现的淋巴管内皮细胞特异性抗原。此外，宿主基因。与编码白介素6、血管内皮生长因子和碱性成纤维细胞生长因子的基因一样，已知在KS皮损中高表达的那些基因在LTC来源的肿瘤中也被诱导表达。KSHV感染的LTCs是第一个KS异种移植模型，可用于KS发病机制的研究和抗KS候选药物的验证。
Kaposi's sarcoma-associated herpesvirus (KSHV) is associated with Kaposi's sarcoma (KS) primary effusion lymphoma (PEL), and multicentric Castleman's disease. Most KS tumor cells are latently infected with KSHV and are of endothelial origin. While PEL-derived cell lines maintain KSHV indefinitely, all KS tumor-derived cells to date have lost viral genomes upon ex vivo cultivation. To study KSHV latency and tumorigenesis in endothelial cells, we generated telomerase-immortalized human umbilical rein endothelial (TIVE) cells. TIVE cells express all KSHV latent genes 48 h postinfection, and productive lytic replication could be induced by, RTA/Orf50. Similar to prior models, infected cultures gradually lost viral episomes. However. we also obtained, for the first time, two endothelial cell lines in which KSHV episomes were maintained indefinitely in the absence of selection. Long-term KSHV maintenance correlated with loss of reactivation in response to RTA-Orf50 and complete oncogenic transformation. Long-term-infected TIVE cells (LTC) grew in soft agar and proliferated under reduced-serum conditions. LTC, but not parental TIVE cells, formed tumors in nude mice. These tumors expressed high levels of the latency-associated nuclear antigen (LANA) and expressed lymphatic endothelial specific antigens as found in KS (LYVE-1). Furthermore, host genes. like those encoding interleukin 6, vascular endothelial growth factor, and basic fibroblast growth factor, known to be highly expressed in KS lesions were also induced in LTC-derived tumors. KSHV-infected LTCs represent the first xenograft model for KS and should be of use to study KS pathogenesis and for the validation of anti-KS drug candidates.