A 50-kDa membrane protein mediates sialic acid-independent binding and infection of conjunctival cells by adenovirus type 37
A 50-kDa membrane protein mediates sialic acid-independent binding and infection of conjunctival cells by adenovirus type 37
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DOI：
10.1006/viro.2000.0703
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发表时间：
2001-01-05
期刊：
影响因子：
3.7
通讯作者：
Nemerow, GR
中科院分区：
文献类型：
--
作者：
Wu, E;Fernandez, J;Nemerow, GR
腺病毒37型(AD37)的嗜眼性与46 kDa柯萨奇病毒和腺病毒受体(CAR)的广泛分布无关，CAR是大多数腺病毒血清型的主要受体。我们先前发现AD37能很好地感染和结合结膜细胞(Chang C)，但不能与表达CAR的肺上皮细胞(A549)结合，并假设该血清型使用一种独特的受体，该受体选择性地表达在结膜细胞上。为了测试这一点，我们制造了含有AD37纤维蛋白的纤维缺失Ad5载体的颗粒。伪型“载体通过非CAR途径感染Chang C细胞，比感染A549细胞效果好。AD37的结合是钙依赖的，并被细胞表面蛋白的蛋白酶消化所取消。利用病毒重叠蛋白印迹分析(VOPBA)，我们检测到钙依赖的AD37与50和60 kDa的膜蛋白结合。相反，在不允许的A549细胞上只检测到60 kDa的蛋白与钙依赖的结合。Ad19p是一种与结膜细胞结合失败的密切相关的血清型，它能识别60 kDa的蛋白，但不能识别50 kDa的蛋白。据报道，AD37用唾液酸代替CAR作为A549细胞的细胞受体。神经氨酸酶可阻断AD37与60 kDa蛋白的结合，提示唾液酸介导了AD37与60 kDa蛋白的结合。假型AD37载体也能感染神经氨酸酶处理的Chang C细胞。因此，D亚群腺病毒与50 kDa蛋白的结合是钙依赖的，并具有细胞类型和血清型特异性，而与60 kDa蛋白的结合不是结膜细胞感染所必需的。在一起。这些数据表明，50 kDa蛋白是结膜细胞上AD37的主要受体。(C)2001年学术出版社。
The ocular tropism of adenovirus type 37 (Ad37) does not correlate with the wide distribution of the 46-kDa coxsackievirus and adenovirus receptor (CAR), the major receptor for most adenovirus serotypes. We previously found that Ad37 infects and binds well to conjunctival cells (Chang C), but poorly to lung epithelial (A549) cells that express CAR and hypothesized that this serotype uses a distinct receptor that is selectively expressed on conjunctival cells. To test this, we produced particles of a fiber-deleted Ad5 vector containing the Ad37 fiber protein. The pseudotyped" vector infected Chang C cells better than A549 cells using a CAR-independent pathway. Ad37 binding was calcium-dependent and was abolished by protease digestion of cell surface proteins. Using a virus overlay protein blot assay (VOPBA), we detected calcium-dependent Ad37 binding to 50- and 60-kDa membrane proteins on permissive Chang C cells. In contrast, calcium-dependent binding was detected with only the 60-kDa protein on nonpermissive A549 cells. Ad19p, a closely related serotype that failed to bind to conjunctival cells, recognized the 60-kDa, but not the 50-kDa, protein. Ad37 has been reported to use sialic acid instead of CAR as a cell receptor on A549 cells. Pretreatment of Chang C cells with neuraminidase abolished Ad37 binding to only the 60-kDa protein, suggesting that sialic acid mediates Ad37 binding to the 60-kDa protein. The pseudotyped Ad37 vector was also able to infect neuraminidase-treated Chang C cells. Thus, subgroup D adenoviral binding to the 50-kDa protein is calcium-dependent and cell type- and serotype-specific, whereas binding to the 60-kDa protein is not necessary for infection of conjunctival cells. Together. these data suggest that the 50-kDa protein is the major receptor for Ad37 on conjunctival cells. (C) 2001 Academic Press.