Enterovirus 71 viral capsid protein linear epitopes: identification and characterization.
Enterovirus 71 viral capsid protein linear epitopes: identification and characterization.
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DOI：
10.1186/1743-422x-9-26
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发表时间：
2012-01-20
期刊：
影响因子：
4.8
通讯作者：
Wang JZ
中科院分区：
文献类型：
--
作者：
Gao F;Wang YP;Mao QY;Yao X;Liu S;Li FX;Zhu FC;Yang JY;Liang ZL;Lu FM;Wang JZ
研究人类对肠病毒71型（EV71）感染的体液免疫反应，并在病毒衣壳蛋白上绘制人类抗原表位。合成了横跨BJ08菌株（基因组C4）衣壳蛋白（VP1, VP2, VP3）的256个肽段。采用间接酶联免疫吸附试验（ELISA）检测患儿急性期和恢复期血清中抗ev71型病毒IgM和IgG的含量。部分重叠肽含有12个氨基酸，作为抗原包被于板上（0.1 μg/μl）。用灭活BJ08病毒免疫家兔血清筛选多肽面板。在急性期血清中共鉴定出10个人抗ev71 IgM表位（VP1中的VP1 -14， VP2中的VP2 -6、21、40和50，VP3中的VP3 -10、12、15、24和75）。而恢复期血清中仅检测到VP1抗ev71 IgG表位（VP1 -15）。4个兔抗ev71 IgG表位（VP1 - 14,31,54和71）被鉴定并定位到VP1上。这些数据表明，人类IgM表位主要定位于VP2和VP3，急性感染时发生多表位反应，而位于VP1蛋白上的IgG表位在恢复期血清中被激活。不同感染阶段体液免疫反应的动态变化可能对EV71疫苗免疫原性评价和诊断试剂的临床应用具有公共卫生意义。
To characterize the human humoral immune response against enterovirus 71 (EV71) infection and map human epitopes on the viral capsid proteins. A series of 256 peptides spanning the capsid proteins (VP1, VP2, VP3) of BJ08 strain (genomic C4) were synthesized. An indirect enzyme-linked immunosorbent assay (ELISA) was carried out to detect anti-EV71 IgM and IgG in sera of infected children in acute or recovery phase. The partially overlapped peptides contained 12 amino acids and were coated in the plate as antigen (0.1 μg/μl). Sera from rabbits immunized with inactivated BJ08 virus were also used to screen the peptide panel. A total of 10 human anti-EV71 IgM epitopes (vp1-14 in VP1; vp2-6, 21, 40 and 50 in VP2 and vp3-10, 12, 15, 24 and 75 in VP3) were identified in acute phase sera. In contrast, only one anti-EV71 IgG epitope in VP1 (vp1-15) was identified in sera of recovery stage. Four rabbit anti-EV71 IgG epitopes (vp1-14, 31, 54 and 71) were identified and mapped to VP1. These data suggested that human IgM epitopes were mainly mapped to VP2 and VP3 with multi-epitope responses occurred at acute infection, while the only IgG epitope located on protein VP1 was activated in recovery phase sera. The dynamic changes of humoral immune response at different stages of infection may have public health significance in evaluation of EV71 vaccine immunogenicity and the clinical application of diagnostic reagents.