Human coronavirus 229E binds to CD13 in rafts and enters the cell through caveolae
Human coronavirus 229E binds to CD13 in rafts and enters the cell through caveolae
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DOI：
10.1128/jvi.78.16.8701-8708.2004
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发表时间：
2004-08-01
影响因子：
5.4
通讯作者：
Fujimoto, T
中科院分区：
文献类型：
--
作者：
Nomura, R;Kiyota, A;Fujimoto, T
CD13 是人类冠状病毒 229E (HCoV-229E) 的受体，被确定为人类成纤维细胞中 Triton X-100 抗性膜微结构域的主要成分。将活成纤维细胞与抗 CD13 抗体在冰上孵育，得到均匀分布在细胞表面的点状标记，但在固定前将温度升高至 37°C 导致标记聚集。在大多数细胞中，CD13 的聚集标记与 Caveolin-1 共定位。 HCoV-229E病毒颗粒表现出与抗CD13抗体类似的结合和重新分布模式：在冰上孵育时，病毒均匀地与细胞结合，但在37℃时与caveolin-1共定位；重要的是，该病毒还导致 CD13 被隔离到 Caveolin-1 阳性区域。电子显微镜证实，在 37°C 下孵育后，HCoV-229E 位于小窝孔口附近或处。用甲基 β-环糊精消耗质膜胆固醇可显着减少 HCoV-229E 的重新分布和随后的感染。通过 RNA 干扰敲低 Caveolin-1 也大大减少了 HCoV-229E 感染。结果表明，HCoV-229E首先与Triton X-100抗性微域中的CD13结合，然后通过交联聚集CD13，从而在进入细胞之前到达小凹区域。
CD13, a receptor for human coronavirus 229E (HCoV-229E), was identified as a major component of the Triton X-100-resistant membrane microdomain in human fibroblasts. The incubation of living fibroblasts with an anti-CD13 antibody on ice gave punctate labeling that was evenly distributed on the cell surface, but raising the temperature to 37degreesC before fixation caused aggregation of the labeling. The aggregated labeling of CD13 colocalized with caveolin-1 in most cells. The HCoV-229E virus particle showed a binding and redistribution pattern that was similar to that caused by the anti-CD13 antibody: the virus bound to the cell evenly when incubated on ice but became colocalized with caveolin-1 at 37degreesC; importantly, the virus also caused sequestration of CD13 to the caveolin-1-positive area. Electron microscopy confirmed that HCoV-229E was localized near or at the orifice of caveolae after incubation at 37degreesC. The depletion of plasmalemmal cholesterol with methyl beta-cyclodextrin significantly reduced the HCoV-229E redistribution and subsequent infection. A caveolin-1 knockdown by RNA interference also reduced the HCoV-229E infection considerably. The results indicate that HCoV-229E first binds to CD13 in the Triton X-100-resistant microdomain, then clusters CD13 by cross-linking, and thereby reaches the caveolar region before entering cells.