Impact of Intra- and Interspecies Variation of Occludin on Its Function as Coreceptor for Authentic Hepatitis C Virus Particles
Impact of Intra- and Interspecies Variation of Occludin on Its Function as Coreceptor for Authentic Hepatitis C Virus Particles
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DOI：
10.1128/jvi.00212-11
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发表时间：
2011-08-01
影响因子：
5.4
通讯作者：
von Hahn, Thomas
中科院分区：
文献类型：
--
作者：
Ciesek, Sandra;Westhaus, Sandra;von Hahn, Thomas
丙型肝炎病毒(HCV)在临床感染过程中具有宿主范围窄、个体间变异性高的特点。这两个特征被认为在很大程度上是由于物种之间和个体寄主之间的遗传差异。紧密连接蛋白(OCLN)是丙型肝炎病毒进入宿主细胞所必需的，人和小鼠OCLN之间的差异被认为是丙型肝炎病毒不能感染小鼠的部分原因，从而排除了将其用作方便的小动物模型的可能性。本研究使用Huh-7.5细胞系的新产生和特征的OCLN(低)亚克隆(其中内源性OCLN的表达被短发夹状RNA下调的Huh-7.5亚克隆)来评估OCLN的遗传变异对细胞培养生长的丙型肝炎病毒(HCVcc)的影响。我们报道了人类群体中编码非同义单核苷酸多态的频率，即导致氨基酸交换的多态，并确定了它们作为丙型肝炎病毒(Co)受体的能力。此外，我们发现小鼠OCLN可以维持HCVcc的进入，尽管与人OCLN相比效率降低了约5倍。这种效率的降低完全是由于以前其他人使用丙型肝炎病毒伪品方法鉴定的两个氨基酸残基。最后，我们使用Huh-7.5/OCLN(LOW)细胞株来表明，丙型肝炎病毒在相邻细胞之间的传播严格依赖于OCLN。
Hepatitis C virus (HCV) is characterized by a narrow host range and high interindividual variability in the clinical course of infection. Both of these traits are thought to be largely due to genetic variation between species and between individual hosts. The tight junction component occludin (OCLN) is essential for HCV entry into host cells, and the differences between human and murine OCLN are thought to account in part for the inability of HCV to infect mice and hence preclude their use as a convenient small-animal model. This study assesses the impact of genetic variation in OCLN on cell culture-grown HCV (HCVcc) using a newly generated and characterized OCLN(low) subclone of the Huh-7.5 cell line (Huh-7.5 subclone in which endogenous OCLN expression has been downregulated by a short hairpin RNA). We report the frequency of coding nonsynonymous single nucleotide polymorphisms, i.e., polymorphisms resulting in amino acid exchanges, present in the human population and determine their ability to function as HCV (co)receptors. Moreover, we show that murine OCLN can sustain HCVcc entry, albeit with about 5-fold reduced efficiency compared to that of human OCLN. This reduction in efficiency is due solely to two amino acid residues previously identified by others using an HCV pseudoparticle approach. Finally, we use the Huh-7.5/OCLN(low) cell line to show that HCV spread between neighboring cells is strictly dependent on OCLN.