Heparan sulfate glycosaminoglycans are receptors sufficient to mediate the initial binding of adenovirus types 2 and 5
Heparan sulfate glycosaminoglycans are receptors sufficient to mediate the initial binding of adenovirus types 2 and 5
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DOI：
10.1128/jvi.75.18.8772-8780.2001
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发表时间：
2001-09-01
影响因子：
5.4
通讯作者：
Cabrini, G
中科院分区：
文献类型：
--
作者：
Dechecchi, MC;Melotti, P;Cabrini, G
腺病毒血清型2和血清型5 （Ad2/5）的细胞感染开始于Ad纤维附着在柯萨奇病毒和Ad受体（CAR）上，然后是α (v)整合素介导的进入。我们最近证实在细胞表面表达的硫酸肝素糖胺聚糖（HS GAGs）参与了Ad2/5的结合和感染（m.c. Dechecchi， A. Tamanini， A. Bonizzato，和G. Cabrini, Virology 268:382- 339,2000）。利用CAR的胞外可溶性结构域1 （CAR- d1）和肝素作为CAR和HS GAGs的可溶性受体类似物，在A549和重组CHO细胞系中研究了HS GAGs的作用，这两种受体的表达水平不同，培养密度不同。Ad2/5与肝素（10马克杯/毫升）和sCAR-D1（200马克杯/毫升）在A549细胞中预孵生，获得了完全的结合和感染抑制。肝素和sCAR-D1分别与Ad预孵育时，观察到部分抑制作用。稀疏A549细胞中肝素敏感[H-3]Ad2/5结合水平（50 ~ 70000细胞/cm(2)）比融合生长细胞（200 ~ 300000细胞/cm(m2)）增加了一倍，同时HS gag表达增加。[H-3]Ad2结合到表达HS gag （CHO K1）的稀疏car阴性CHO细胞上。在CHO KI细胞和HS gag缺陷的CHO A745、D677和E606克隆中，通过肝素的竞争性抑制，观察到No [H-3]Ad2结合。在CAR阴性的稀疏的CHO K1细胞中发现了hs敏感的Ad2感染，但在CHO A745细胞中没有，CHO A745细胞只有在转染CAR后才允许感染。这些结果表明，HS gag足以介导Ad2/5的初始结合。
Cell infection by adenovirus serotypes 2 and 5 (Ad2/5) initiates with the attachment of Ad fiber to the coxsackievirus and Ad receptor (CAR) followed by alpha (v) integrin-mediated entry. We recently demonstrated that heparan sulfate glycosaminoglycans (HS GAGs) expressed on cell surfaces are involved in the binding and infection of Ad2/5 (M. C. Dechecchi, A. Tamanini, A. Bonizzato, and G. Cabrini, Virology 268:382-390, 2000). The role of HS GAGs was investigated using extracellular soluble domain 1 of CAR (sCAR-D1) and heparin as soluble receptor analogues of CAR and HS GAGs in A549 and recombinant CHO cell lines with differential levels of expression of the two receptors and cultured to various densities. Complete inhibition of binding and infection was obtained by preincubating Ad2/5 with both heparin (10 mug/ml) and sCAR-D1 (200 mug/ml) in A549 cells. Partial inhibition was observed when heparin and sCAR-D1 were preincubated separately with Ad. The level of heparin-sensitive [H-3]Ad2/5 binding doubled in sparse A549 cells (50 to 70,000 cells/cm(2)) with respect to that of cells grown to confluence (200 to 300,000 cells/cm2), in parallel with increased expression of HS GAGs. [H-3]Ad2 bound to sparse CAR-negative CHO cells expressing HS GAGs (CHO K1). No [H-3]Ad2 binding was observed in CHO KI cells upon competitive inhibition with heparin and in HS GAG-defective CHO A745, D677, and E606 clones. HS-sensitive Ad2 infection was obtained in CAR-negative sparse CHO K1 cells but not in CHO A745 cells, which were permissive to infection only upon transfection with CAR. These results demonstrate that HS GAGs are sufficient to mediate the initial binding of Ad2/5.