Antigenic mapping of the recombinant Norwalk virus capsid protein using monoclonal antibodies
Antigenic mapping of the recombinant Norwalk virus capsid protein using monoclonal antibodies
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DOI：
10.1006/viro.1996.0112
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发表时间：
1996-03-01
期刊：
影响因子：
3.7
通讯作者：
Estes, MK
中科院分区：
文献类型：
--
作者：
Hardy, ME;Tanaka, TN;Estes, MK
诺瓦克病毒（Norwalk virus，NV）是一类不可培养的杯状病毒的原型毒株，可感染人类并引起急性非细菌性胃肠炎的暴发流行。NV病毒体由180个拷贝的单一结构蛋白组成，当在用重组杆状病毒感染的昆虫细胞中表达时，所述单一结构蛋白组装成空的重组诺瓦克病毒样颗粒（rNV VLP），所述空的重组诺瓦克病毒样颗粒在形态学和抗原性上与天然NV相似。我们已经开始使用针对rNV VLP制备的单克隆抗体来剖析rNV颗粒的抗原结构。针对rNV颗粒制备的10种MAb的反应性通过ELISA表征为检测抗体，在ELISA中表征为捕获抗体以检测粪便中的NV，通过Western印迹和通过免疫沉淀表征。七个单克隆抗体识别不连续的表位，需要rNV衣壳蛋白保持至少部分折叠，而其他三个识别连续的表位。八个单克隆抗体映射到C-末端一半的衣壳蛋白，因为它们通过Western印迹和通过免疫沉淀与全长58 K衣壳蛋白的32 K胰蛋白酶切割产物反应，表明C-末端一半的衣壳蛋白可能含有免疫显性表位。识别连续表位的三种单克隆抗体映射到衣壳蛋白的最末端C末端，在氨基酸457和530之间，在不同人杯状病毒衣壳蛋白之间相对保守的区域中。这些单克隆抗体被分配到三个抗原组，将是有用的工具，以进一步剖析NV病毒粒子的结构和抗原拓扑结构，并作为无限的试剂，以检测NV的诊断测定。(C)出版社：Academic Press，Inc.
Norwalk virus (NV) is the prototype strain of a group of noncultivatable caliciviruses that infect humans and cause outbreaks of epidemic acute nonbacterial gastroenteritis. The NV virion is composed of 180 copies of a single structural protein that, when expressed in insect cells infected with a recombinant baculovirus, assembles into empty recombinant Norwalk virus-like particles (rNV VLPs) which are morphologically and antigenically similar to native NV. We have begun to dissect the antigenic structure of the rNV particles using monoclonal antibodies made to the rNV VLPs. Ten MAbs made to rNV particles were characterized for their reactivity as detector antibodies by ELISA, as capture antibodies in an ELISA to detect NV in stools, by Western blot, and by immunoprecipitation. Seven of the MAbs recognize discontinuous epitopes, requiring the rNV capsid protein to remain at least partially folded, while the other three recognize continuous epitopes. Eight of the MAbs map to the C-terminal half of the capsid protein as they react by Western blot and by immunoprecipitation with a 32K trypsin cleavage product of the full-length 58K capsid protein, suggesting that the C-terminal half of the capsid protein may contain the immunodominant epitopes. The three MAbs that recognize continuous epitopes map to the extreme C terminus of the capsid protein, between amino acids 457 and 530, in a region that is relatively conserved among different human calicivirus capsid proteins. These MAbs which were assigned into three antigenic groups will be useful as tools to further dissect the structural and antigenic topography of the NV virion, and as unlimited reagents to detect NV in diagnostic assays. (C) 1996 Academic Press, Inc.