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通过代表 T 辅助细胞和 B 细胞表位的嵌合肽诱导针对西尼罗河病毒和日本脑炎病毒的特异性中和免疫应答。

Induction of virus-specific neutralizing immune response against West Nile and Japanese encephalitis viruses by chimeric peptides representing T-helper and B-cell epitopes.

机构信息

Encephalitis Group, National Institute of Virology, Sus Road Campus, 130/1, Pashan, Pune 411021, India.

出版信息

Virus Res. 2012 Jan;163(1):40-50. doi: 10.1016/j.virusres.2011.08.008. Epub 2011 Aug 27.

Abstract

West Nile virus (WNV) and Japanese encephalitis virus (JEV), the members of JEV serocomplex group are pathogens of global health concern. The co-circulation of these viruses poses challenges in effective diagnostics due to antigenic similarity between the E-protein of these viruses. The present study aimed to design chimeric peptides and study the immune response against the same. B-cell epitopes were predicted on structural proteins of WNV and JEV based on bioinformatics tools. The peptides representing to these B-cell epitopes were synthesized and subjected to ELISA. Two peptides, one each from WNV (named WE147) and JEV (named JE40) E-protein, showed virus-specific and strong reactivity to the immune mice sera and human clinical samples. The chimeric peptides for WNV and JEV were constructed by synthesizing the B-cell epitope of WNV (WE147) or JEV (JE40) with T-helper epitope (JM17) separated by diglycine spacer in between. The immune response generated against these chimeric peptides was found to be specific to the respective B-cell epitopes. The anti-peptide sera showed virus-specific reactivity in ELISA and in immunofluorescence assay with no cross-reactivity. Also, the anti-peptide sera could neutralize JE and WN viruses in an in vitro virus neutralization assay. The B-cell epitopes identified in the present study may be used as diagnostic markers for differentiating between WN and JE virus infections. The present study can form a basis for future design of vaccines.

摘要

西尼罗河病毒(WNV)和日本脑炎病毒(JEV)是 JEV 血清复合物群的成员,是全球健康关注的病原体。由于这些病毒的 E 蛋白之间存在抗原相似性,因此这些病毒的共同循环给有效诊断带来了挑战。本研究旨在设计嵌合肽并研究针对这些肽的免疫反应。根据生物信息学工具,预测了 WNV 和 JEV 结构蛋白上的 B 细胞表位。合成了代表这些 B 细胞表位的肽,并进行了 ELISA 分析。两种肽,一种来自 WNV(命名为 WE147),另一种来自 JEV(命名为 JE40)的 E 蛋白,对免疫小鼠血清和人类临床样本表现出特异性和强反应。通过合成 WNV(WE147)或 JEV(JE40)的 B 细胞表位,并用二肽间隔子将 T 辅助表位(JM17)分隔开来,构建了针对 WNV 和 JEV 的嵌合肽。针对这些嵌合肽产生的免疫反应被发现是针对各自 B 细胞表位的特异性反应。抗肽血清在 ELISA 和免疫荧光测定中表现出针对病毒的特异性反应,没有交叉反应。此外,抗肽血清可以在体外病毒中和测定中中和 JE 和 WN 病毒。本研究中鉴定的 B 细胞表位可用作区分 WN 和 JE 病毒感染的诊断标志物。本研究可以为未来的疫苗设计奠定基础。

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