Hepatitis B virus infection is dependent on cholesterol in the viral envelope
Hepatitis B virus infection is dependent on cholesterol in the viral envelope
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DOI：
10.1111/j.1462-5822.2008.01250.x
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发表时间：
2009-02-01
影响因子：
3.4
通讯作者：
Glebe, Dieter
中科院分区：
文献类型：
--
作者：
Bremer, Corinna M.;Bung, Christiane;Glebe, Dieter
导致B型肝炎病毒（HBV）结合和进入的病毒和细胞决定簇仍然没有完全理解。我们发现，HBV感染的原代肝细胞培养是依赖于胆固醇的存在下，在病毒包膜。用甲基-β-环糊精（M β CD）从HBV感染患者血浆中纯化的HBV中提取胆固醇，可大大降低感染水平。胆固醇耗尽的病毒粒子显示出更高的浮力密度（1.23比1.17 g ml（-1）），更小的直径（39比48 nm），但保持颗粒完整性，抗原性和结合肝细胞的能力。虽然添加外源性胆固醇和胆固醇类似物恢复了胆固醇耗尽的病毒粒子的物理外观，但只有通过胆固醇回加才能恢复感染性。在存在胆固醇合成抑制剂的情况下，从细胞培养物产生的HBV的免疫力严重受损。有趣的是，从细胞膜中提取胆固醇，与菲律宾蛋白和蛋白酪氨酸激酶抑制剂染料木黄酮孵育显示对HBV感染没有影响，排除了脂筏在HBV感染过程中的作用。总之，病毒包膜内胆固醇的存在对于病毒结合并不重要，但对于HBV的进入过程是必不可少的，并且对于病毒摄取的后续步骤（例如在未知隔室中融合）可能是重要的。
The viral and cellular determinants leading to binding and entry of hepatitis B virus (HBV) are still not fully understood. We found that HBV infection of primary hepatocyte cultures is dependent on the presence of cholesterol in the viral envelope. Extraction of cholesterol from HBV purified from plasma of HBV-infected patients with methyl-beta-cyclodextrin (M beta CD) leads to a strongly reduced level of infection. The cholesterol-depleted virions showed higher buoyant density (1.23 versus 1.17 g ml(-1)), a smaller diameter (39 versus 48 nm), but maintained particle integrity, antigenicity and ability to bind to hepatocytes. Although addition of exogenous cholesterol and cholesterol analogues restored the physical appearance of cholesterol-depleted virions, infectivity was only regained by cholesterol add-back. Infectivity of HBV produced from cell culture in the presence of inhibitors of cholesterol-synthesis is severely impaired. Interestingly, cholesterol extraction from cellular membranes, incubation with filipin and the protein tyrosine kinase inhibitor genistein showed no effect on HBV infection, excluding a role of lipid rafts for the infection process of HBV. In summary, presence of cholesterol within the viral envelope is not important for viral binding, but indispensable for the entry process of HBV and might be important for a later step in viral uptake, e.g. fusion in a yet unknown compartment.