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通过针对基质蛋白的单克隆抗体所定义的北美和欧洲猪繁殖与呼吸综合征病毒株之间的抗原变异性。

Antigenic variability among North American and European strains of porcine reproductive and respiratory syndrome virus as defined by monoclonal antibodies to the matrix protein.

作者信息

Dea S, Gagnon C A, Mardassi H, Milane G

机构信息

Centre de Recherche en Virologie, Institut Armand-Frappier, Université du Québec, Laval, Canada.

出版信息

J Clin Microbiol. 1996 Jun;34(6):1488-93. doi: 10.1128/jcm.34.6.1488-1493.1996.

Abstract

Two hybridoma cell lines producing monoclonal antibodies (Mabs) to the 19-kDa matrix (M) protein of porcine reproductive and respiratory syndrome virus (PRRSV) were obtained from BALB/c mice that were immunized with a reference Quebec tissue culture-adapted strain (strain IAF-Klop). The polypeptide specificities of the MAbs were determined by immunoblotting and immunoprecipitation tests with concentrated and purified preparations of the virus and by determining their reactivities with the Escherichia coli-expressed gene products of open reading frames 5 to 7. The two anti-M protein MAbs (MAbs IAFK3 and IAFK6) and another MAb (MAb IAFK8) directed to the 15-kDa nucleocapsid (N) protein were devoid of virus-neutralizing activity. A library of four anti-N protein MAbs (MAbs IAFK8, SDOW17, VO17, and EP147) and two anti-M protein MAbs (MAbs IAFK6 and IAFK3) was used to investigate, by an indirect fluorescent-antibody assay, the antigenic diversity of 15 Canadian PRRSV isolates, in comparison with those of the U.S. ATCC VR2332 attenuated vaccine strain and two reference European (Lelystad and Weybridge) PRRSV strains. The North American and European PRRSV isolates tested shared the epitopes recognized by anti-N protein MAbs IAFK8 and SDOW17, but three distinct patterns could be identified on the basis of their reactivities with the other anti-PRRSV MAbs. No reactivity to the anti-M protein MAbs was observed by either European PRRSV isolate or the attenuated U.S. vaccine strain.

摘要

从用魁北克参考组织培养适应株(IAF-Klop株)免疫的BALB/c小鼠中获得了两个产生针对猪繁殖与呼吸综合征病毒(PRRSV)19-kDa基质(M)蛋白的单克隆抗体(Mab)的杂交瘤细胞系。通过对浓缩和纯化的病毒制剂进行免疫印迹和免疫沉淀试验,并通过测定它们与大肠杆菌表达的开放阅读框5至7的基因产物的反应性,确定了这些单克隆抗体的多肽特异性。两种抗M蛋白单克隆抗体(单克隆抗体IAFK3和IAFK6)以及另一种针对15-kDa核衣壳(N)蛋白的单克隆抗体(单克隆抗体IAFK8)没有病毒中和活性。使用一个包含四种抗N蛋白单克隆抗体(单克隆抗体IAFK8、SDOW17、VO17和EP147)和两种抗M蛋白单克隆抗体(单克隆抗体IAFK6和IAFK3)的文库,通过间接荧光抗体试验,与美国ATCC VR2332减毒疫苗株以及两种欧洲参考PRRSV株(莱利斯塔德株和韦布里奇株)相比,研究了15株加拿大PRRSV分离株的抗原多样性。所测试的北美和欧洲PRRSV分离株共享抗N蛋白单克隆抗体IAFK8和SDOW17识别的表位,但根据它们与其他抗PRRSV单克隆抗体的反应性可识别出三种不同模式。欧洲PRRSV分离株或美国减毒疫苗株均未观察到与抗M蛋白单克隆抗体的反应性。

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