Characterization of monoclonal antibodies to the hemagglutinin-esterase glycoprotein of a bovine coronavirus associated with winter dysentery and cross-reactivity to field isolates
Characterization of monoclonal antibodies to the hemagglutinin-esterase glycoprotein of a bovine coronavirus associated with winter dysentery and cross-reactivity to field isolates
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DOI：
10.1128/jcm.35.1.33-40.1997
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发表时间：
1997-01-01
影响因子：
9.4
通讯作者：
Dea, S
中科院分区：
文献类型：
--
作者：
Milane, G;Kourtesis, AB;Dea, S
用冬痢（WD）相关菌株BCQ.2590富集聚体部分免疫BALB/c小鼠，获得7株产生牛冠状病毒（BCV）血凝素-酯酶（HE）糖蛋白单克隆抗体（mab）的杂交瘤细胞株。这些单克隆抗体对二聚体（140-kDa）或单体（65-kDa）形式的HE糖蛋白的特异性通过纯化病毒的免疫印迹实验和[S-35]蛋氨酸标记的感染细胞提取物的免疫沉淀试验来确定。这些抗he单克隆抗体中有四种抑制病毒的血凝活性，三种在体外弱中和其传染性。此外，竞争结合分析允许为wd相关菌株的HE糖蛋白定义两个独立的抗原结构域（结构域A和D）和两个重叠的抗原结构域（结构域B和C）；抗原结构域A内的表位与血凝抑制（HAI）和病毒中和活性无关。在HAI试验中，四种抗ha单克隆抗体在24个BCV分离株中定义了两个不同的抗原亚群，这些分离株与1986年至1996年魁北克奶牛群中典型的WD或新生儿小牛腹泻（NCD）暴发有关。魁北克省登革热相关的BCV毒株，以及自1991年以来分离的一些非传染性疾病相关毒株，属于与Mebus原型毒株不同的一个亚群。
Seven hybridoma cell lines producing monoclonal antibodies (MAbs) to the hemagglutinin-esterase (HE) glycoprotein of bovine coronavirus (BCV) were obtained from BALB/c mice that were immunized with an enriched peplomeric fraction of the winter dysentery (WD)-associated strain BCQ.2590. The specificities of these MAbs to either the dimeric (140-kDa) or the monomeric (65-kDa) form of the HE glycoprotein were determined by Western immunoblotting experiments with purified virus and immunoprecipitation tests with [S-35] methionine-labelled infected cell extracts. Four of these anti-HE MAbs inhibited the hemagglutinating activity of the virus and three weakly neutralized its infectivity in vitro. In addition, competition binding assays allowed for the definition of two independent antigenic domains (domains A and D) and two overlapping antigenic domains (domains B and C) for the HE glycoprotein of the WD-associated strain; epitopes located within antigenic domain A were not associated with hemagglutination inhibition (HAI) and virus neutralization activities. In HAI tests, the four anti-HA MAbs defined two distinct antigenic subgroups among 24 BCV field isolates that have been associated with either typical outbreaks of WD or neonatal calf diarrhea (NCD) in Quebec dairy herds from 1986 to 1996. The Quebec WD-associated strains of BCV, as well as some of the NCD-associated strains isolated since 1991, fell within a subgroup distinct from that of the prototype Mebus strain.