Qualitative and Quantitative Analysis of the Binding of GII.4 Norovirus Variants onto Human Blood Group Antigens
Qualitative and Quantitative Analysis of the Binding of GII.4 Norovirus Variants onto Human Blood Group Antigens
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DOI：
10.1128/jvi.02077-10
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发表时间：
2011-05-01
影响因子：
5.4
通讯作者：
Belliot, G.
中科院分区：
文献类型：
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作者：
de Rougemont, A.;Ruvoen-Clouet, N.;Belliot, G.
诺如病毒（NoV）是儿童和成人胃肠炎的主要原因之一。在过去的20年中，基因组II基因型4（GII.4）NoV已经在世界范围内传播。GII.4 NoV可分为多种变体，自2002年以来，它们在每2或3年更换一次之前在人群中传播，这引起了关于其组织血型抗原（HBGA）配体在其进化中的作用的问题。为了阐明这些问题，我们对代表性GII.4变体和HBGAs之间的相互作用进行了分析，并确定了所选氨基酸在结合谱中的作用。通过诱变，我们表明，有一个严格的结构要求的氨基酸，直接涉及与HBGAs的相互作用。然而，在位置395处的苏氨酸残基（Δ T395）的消融（2002年后变体的流行病学特征）对病毒样颗粒（VLP）与H抗原的结合无害，而与A和B抗原的结合严重受阻。然而，与野生型VLP相比，Delta T395 VLP获得了与刘易斯x和唾液酸-刘易斯x抗原结合的能力，表明HBGA结合位点外的氨基酸残基可以改变NoV的结合性质。我们还分析了从1987年至2007年分离的6种变异体（布里斯托、US 95/96、Hunter、Yerseke、Den Haag和Osaka）中杆状病毒表达的VLP与表型唾液样本和合成HBGA的附着。我们发现，这六种变体都可以附着在分泌者的唾液上，而不管ABO表型和分泌者表型的寡糖特征。有趣的是，海牙和大坂变异体还与刘易斯阳性非分泌者唾液中存在的碳水化合物结合。碳水化合物结合特征以及遗传和诱变分析表明，GII. 4与刘易斯x和唾液酸-刘易斯x抗原的结合可能是位于结合位点附近的氨基酸遗传变异的副产物。通过表面等离子体共振对六种变体的结合特性的分析表明，只有2002年后的变体（即，Hunter、Yerseke、Den Haag和Osaka）与A和B抗原的结合力很强，这表明GII. 4进化可能与2002年后变异体对HBGA的亲和力增加有关。ABH抗原的亲和力增加和新获得的识别来自路易斯阳性非分泌者的聚糖的能力的结合可能有助于菌株如海牙GII.4亚型的流行病学重要性。
Noroviruses (NoVs) are one of the leading causes of gastroenteritis in children and adults. For the last 2 decades, genogroup II genotype 4 (GII.4) NoVs have been circulating worldwide. GII.4 NoVs can be divided into variants, and since 2002 they have circulated in the population before being replaced every 2 or 3 years, which raises questions about the role of their histo-blood group antigen (HBGA) ligands in their evolution. To shed light on these questions, we performed an analysis of the interaction between representative GII.4 variants and HBGAs, and we determined the role of selected amino acids in the binding profiles. By mutagenesis, we showed that there was a strict structural requirement for the amino acids, directly implicated in interactions with HBGAs. However, the ablation of the threonine residue at position 395 (Delta T395), an epidemiological feature of the post-2002 variants, was not deleterious to the binding of the virus-like particle (VLP) to the H antigen, while binding to A and B antigens was severely hampered. Nevertheless, the Delta T395 VLPs gained the capacity to bind to the Lewis x and sialyl-Lewis x antigens in comparison with the wild-type VLP, demonstrating that amino acid residues outside the HBGA binding site can modify the binding properties of NoVs. We also analyzed the attachment of baculovirus-expressed VLPs from six variants (Bristol, US95/96, Hunter, Yerseke, Den Haag, and Osaka) that were isolated from 1987 to 2007 to phenotyped saliva samples and synthetic HBGAs. We showed that the six variants could all attach to saliva of secretors irrespective of the ABO phenotype and to oligosaccharides characteristic of the secretor phenotype. Interestingly, Den Haag and Osaka variants additionally bound to carbohydrates present in the saliva of Lewis-positive nonsecretors. The carbohydrate binding profile and the genetic and mutagenesis analysis suggested that GII.4 binding to Lewis x and sialyl-Lewis x antigens might be a by-product of the genetic variation of the amino acids located in the vicinity of the binding site. Analysis of the binding properties for the six variants by surface plasmon resonance showed that only post-2002 variants (i.e., Hunter, Yerseke, Den Haag, and Osaka) presented strong binding to A and B antigens, suggesting that the GII.4 evolution could be related to an increased affinity for HBGAs for the post-2002 variants. The combination of increased affinity for ABH antigens and of a newly acquired ability to recognize glycans from Lewis-positive nonsecretors could have contributed to the epidemiological importance of strains such as the Den Haag GII.4 subtype.