Hepatitis B and D Viruses Exploit Sodium Taurocholate Co-transporting Polypeptide for Species-Specific Entry into Hepatocytes
Hepatitis B and D Viruses Exploit Sodium Taurocholate Co-transporting Polypeptide for Species-Specific Entry into Hepatocytes
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DOI：
10.1053/j.gastro.2013.12.024
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发表时间：
2014-04-01
期刊：
影响因子：
29.4
通讯作者：
Urban, Stephan
中科院分区：
文献类型：
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作者：
Ni, Yi;Lempp, Florian A.;Urban, Stephan
背景与目的：乙型和丁型肝炎病毒（HBV和HDV）是一种宿主范围有限、对肝细胞具有高选择性的人类病原体；HBV l -包膜蛋白与这些细胞上的受体特异性相互作用。我们的目的是鉴定这种受体，并分析它是否是最近描述的由SLC10A1基因编码的牛磺酸胆酸钠共转运蛋白多肽（NTCP）。方法：为了鉴定候选受体，我们比较了表达受体的分化HepaRG细胞和不表达受体的幼稚细胞之间的基因表达模式。在HepaRG细胞中通过小发夹RNA沉默来评估候选受体；在转导的肝癌细胞系中测试了受体表达赋予结合和感染的能力。我们使用种间结构域交换来鉴定受体介导的HBV和HDV结合和感染宿主识别的基序。结果：比较表达阵列的生物信息学分析证实，之前通过生化方法鉴定的NTCP是HBV和HDV的真正受体。来自大鼠、小鼠和人的NTCPs结合Myrcludex B，一种从HBV l蛋白衍生的肽配体。Myrcludex B阻断胆汁盐NTCP转运；在HepaRG细胞中，小发夹rna介导的NTCP敲低可防止HBV或HDV的感染。人类而非小鼠NTCP在HepG2和HuH7细胞中的表达赋予了有限的细胞类型相关和病毒依赖的感染易感性；当用二甲亚砜培养细胞时，这些限制被克服了。我们在人类NTCP中发现了2个短序列基序，它们是HBV和HDV的物种特异性结合和感染所必需的。结论：人NTCP是HBV和HDV的特异性受体。表达ncpp的细胞系可以有效地感染这些病毒，并可用于基础研究和高通量筛选研究。nntcp中基序的定位增加了我们对HBV和HDV物种特异性的理解，并可能为研究病毒感染和复制提供小动物模型。
BACKGROUND & AIMS: Hepatitis B and D viruses (HBV and HDV) are human pathogens with restricted host ranges and high selectivity for hepatocytes; the HBV L-envelope protein interacts specifically with a receptor on these cells. We aimed to identify this receptor and analyze whether it is the recently described sodium-taurocholate co-transporter polypeptide (NTCP), encoded by the SLC10A1 gene. METHODS: To identify receptor candidates, we compared gene expression patterns between differentiated HepaRG cells, which express the receptor, and naive cells, which do not. Receptor candidates were evaluated by small hairpin RNA silencing in HepaRG cells; the ability of receptor expression to confer binding and infection were tested in transduced hepatoma cell lines. We used interspecies domain swapping to identify motifs for receptor-mediated host discrimination of HBV and HDV binding and infection. RESULTS: Bioinformatic analyses of comparative expression arrays confirmed that NTCP, which was previously identified through a biochemical approach is a bona fide receptor for HBV and HDV. NTCPs from rat, mouse, and human bound Myrcludex B, a peptide ligand derived from the HBV L-protein. Myrcludex B blocked NTCP transport of bile salts; small hairpin RNA-mediated knockdown of NTCP in HepaRG cells prevented their infection by HBV or HDV. Expression of human but not mouse NTCP in HepG2 and HuH7 cells conferred a limited cell-type-related and virus-dependent susceptibility to infection; these limitations were overcome when cells were cultured with dimethyl sulfoxide. We identified 2 short-sequence motifs in human NTCP that were required for species-specific binding and infection by HBV and HDV. CONCLUSIONS: Human NTCP is a specific receptor for HBV and HDV. NTCP-expressing cell lines can be efficiently infected with these viruses, and might be used in basic research and high-throughput screening studies. Mapping of motifs in NTCPs have increased our understanding of the species specificities of HBV and HDV, and could lead to small animal models for studies of viral infection and replication.