Iduronic acid-containing glycosaminoglycans on target cells are required for efficient respiratory syncytial virus infection
Iduronic acid-containing glycosaminoglycans on target cells are required for efficient respiratory syncytial virus infection
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DOI：
10.1006/viro.2000.0293
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发表时间：
2000-06-05
期刊：
影响因子：
3.7
通讯作者：
Peeples, ME
中科院分区：
文献类型：
--
作者：
Hallak, LK;Collins, PL;Peeples, ME
呼吸道合胞病毒（RSV）是一种重要的人类呼吸道病原体，特别是在婴儿中。糖胺聚糖（GAG）与RSV感染培养细胞的起始有关，但尚不清楚涉及何种类型的GAG和GAG组分，重要的GAG是在病毒上还是在细胞上，或者它们对感染的贡献程度如何。我们构建并拯救了一种表达重组绿色荧光蛋白（GFP）的RSV（rgRSV），并使用该病毒开发了一种敏感的系统，通过流式细胞术评估和定量感染。对一组GAG合成各方面遗传缺陷的突变型中国仓鼠卵巢细胞系进行的评价显示，根据GAG缺陷的类型，感染降低高达80%。从HEp-2细胞表面酶促去除硫酸乙酰肝素和/或硫酸软骨素也减少了感染，去除两者甚至进一步减少了感染。将RSV与各种可溶性GAG预孵育的阻断实验显示相对阻断顺序为：肝素>硫酸乙酰肝素>硫酸软骨素B。艾杜糖醛酸是这些GAG共有的组分，不含艾杜糖醛酸的GAG，即硫酸软骨素A和C以及透明质酸，不抑制感染。碱性成纤维细胞生长因子阻断感染的能力证实了含艾杜糖醛酸的GAG在RSV感染中的作用，因为碱性成纤维细胞生长因子与含艾杜糖醛酸的GAG结合。用硫酸鱼精蛋白预处理细胞，可以结合和阻断GAG，也可以减少感染。在这些实施例中，通过用可溶性GAG预处理病毒、用GAG结合分子预处理细胞、用GAG破坏酶预处理细胞或在GAG遗传缺陷的细胞中减少感染。这些结果证实参与RSV感染的GAG存在于细胞上而不是病毒颗粒上。因此，含有艾杜糖醛酸的细胞表面GAG如硫酸乙酰肝素和硫酸软骨素B的存在是细胞培养物中有效RSV感染所必需的。(C)北京大学出版社.
Respiratory syncytial virus (RSV) is an important human respiratory pathogen, particularly in infants. Glycosaminoglycans (GAGs) have been implicated in the initiation of RSV infection of cultured cells, but it is not clear what type of GAGs and GAG components are involved, whether the important GAGs are on the virus or the cell, or what the magnitude is of their contribution to infection. We constructed and rescued a recombinant green fluorescent protein (GFP)-expressing RSV (rgRSV) and used this virus to develop a sensitive system to assess and quantify infection by flow cytometry. Evaluation of a panel of mutant Chinese hamster ovary cell lines that are genetically deficient in various aspects of GAG synthesis showed that infection was reduced up to 80% depending on the type of GAG deficiency. Enzymatic removal of heparan sulfate and/or chondroitin sulfate from the surface of HEp-2 cells also reduced infection, and the removal of both reduced infection even further. Blocking experiments in which RSV was preincubated with various soluble GAGs revealed the relative blocking order of: heparin > heparan sulfate > chondroitin sulfate B. Iduronic acid is a component common to these GAGs, GAGs that do not contain iduronic acid, namely, chondroitin sulfate A and C and hyaluronic acid, did not inhibit infection. A role for iduronic acid-containing GAGs in RSV infection was confirmed by the ability of basic fibroblast growth factor to block infection, because basic fibroblast growth factor binds to GAGs containing iduronic acid. Pretreatment of cells with protamine sulfate, which binds and blocks GAGs, also reduced infection. In these examples, infection was reduced by pretreatment of the virus with soluble GAGs, pretreatment of the cells with GAG-binding molecules, pretreatment of the cells with GAG-destroying enzymes or in cells genetically deficient in GAGs. These results establish that the GAGs involved in RSV infection are present on the cell rather than on the virus particle. Thus, the presence of cell surface GAGs containing iduronic acid, like heparan sulfate and chondroitin sulfate B, is required for efficient RSV infection in cell culture. (C) 2000 Academic Press.