Characterization of entry mechanisms of human herpesvirus 8 by using an Rta-dependent reporter cell line
Characterization of entry mechanisms of human herpesvirus 8 by using an Rta-dependent reporter cell line
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DOI：
10.1128/jvi.77.14.8147-8152.2003
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发表时间：
2003-07-01
影响因子：
5.4
通讯作者：
Koyano, S
中科院分区：
文献类型：
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作者：
Inoue, N;Winter, J;Koyano, S
为了分析人类疱疹病毒8 （HHV-8）的进入机制，我们建立了一株含有lacZ基因的报告细胞系T1H6，该细胞系在多腺苷化核RNA启动子的控制下，已知该启动子可被病毒反激活子Rta强烈激活。我们发现，感染无细胞病毒，以及与hhv -8阳性原发性积液淋巴瘤细胞系共培养，以敏感和剂量依赖的方式激活T1H6的lacZ基因。聚苯乙烯的加入和离心作用增强了HHV-8的传染性，但聚磺酸化合物抑制了HHV-8的传染性。含有rgd基序的多肽和整合素并没有降低传染性，这表明除了报道的受体外，还存在其他细胞受体。这种进入依赖于pH酸化，而不依赖于网格蛋白途径。虽然从人类免疫缺陷病毒（HIV）感染的细胞中获得的条件培养基对HHV-8感染的早期阶段没有任何影响，但细胞内表达一种前HIV 1型，而不是单独表达Tat，可以略微增加HHV-8依赖性报告基因的激活，这表明HIV介导的增强HHV-8感染的早期阶段的潜力。
To analyze the mechanisms of entry of human herpesvirus 8 (HHV-8), we established a reporter cell line T1H6 that contains the lacZ gene under the control of the polyadenylated nuclear RNA promoter, known to be strongly activated by a viral transactivator, Rta. We found that infection with cell-free virus, as well as cocultivation with HHV-8-positive primary effusion lymphoma cell lines, activated the lacZ gene of T1H6 in a sensitive and dose-dependent manner. Addition of Polybrene and centrifugation enhanced, but polysulfonate compounds inhibited, the HHV-8 infectivity. RGD-motif-containing polypeptides and integrins did not decrease the infectivity, suggesting the presence of an additional cellular receptor other than the reported one. The entry was dependent on pH acidification but not on the clathrin pathway. Although conditioned media obtained from human immunodeficiency virus (HIV)-infected cells did not have any effect on the early steps of HHV-8 infection, intracellular expression of a proviral HIV type 1, but not of Tat alone, increased the HHV-8-dependent reporter activation slightly, suggesting a potential of HIV-mediated enhancement of an early step of HHV-8 infection.