高致病性猪繁殖与呼吸综合征病毒GP5蛋白B表位诱导中和抗体能力的研究  

The ability of B epitope in GP5 protein of highly pathogenic PRRSV to induce neutralizing antibody

作　　者：冷超粮[1] 安同庆[1] 陈家锃[1] 宫大庆[1] 李登云[1] 杨永倩[1] 彭金美[1] 张祎[1] 郭娟娟[1] 吴江[1] 童光志[2] 田志军[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国预防兽医学报》2011年第4期297-300,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家重点基础研究发展计划(973)项目(2005CB523202);兽医生物技术国家重点实验室自主研究项目(SKLVBP201011)

摘　　要：为证实GP5蛋白B表位是否具有诱导抗猪繁殖与呼吸综合征病毒(PRRSV)中和抗体的能力,本实验合成了PRRSV CH-1a株和高致病性PRRSV(HP-PRRSV)HuN4株B表位的短肽,分别免疫家兔制备两份特异性多肽抗血清。此外,将12头PRRSV及其抗体均为阴性的健康仔猪以HP-PRRSV HuN4疫苗毒株(HuN4-F112)进行基础免疫,然后用强毒株(HuN4-F5)进行多次接种,制备猪高免血清。间接免疫荧光检测(IFA)结果显示,两份多肽抗血清均能与PRRSV经典株和变异株发生特异性反应,IFA抗体效价无差异,表明两份血清没有显著差异。病毒中和试验结果表明,两份多肽抗血清均不具有中和PRRSV活性。间接ELISA和病毒中和试验结果表明,猪高免血清中针对B表位肽的ELISA抗体水平与血清抗PRRSV中和抗体之间没有正相关关系,而且B表位肽不能抑制中和抗体。以上结果表明HP-PRRSV B表位不能诱导产生中和抗体。B epitope was considered as a major linear neutralizing epitope in the GP5 protein of porcine reproductive and respiratory syndrome virus （PRRSV） classical strains. Compared with the classical strains, however, one amino acid mutation （L39I） was found in the B epitope of highly pathogenic PRRSV （HP-PRRSV） strains. To study the ability of the B epitope to induce neutralizing antibody （NA） against HP-PRRSV, the B epitope peptides with and without the mutation （L39I） were synthesized and immunized in rabbits to prepare the anti-peptide serums, respectively. Pig hyperimmune serums with high titer of NAs against PRRSV were prepared by hyperimmunization of swine with HP-PRRSV HuN4 vaccine strain （HuN4-F112） and virulent strain （HuN4-F5）. Indirect immunofluorescence assay （IFA） showed that both rabbit anti-peptide serums had positive reactivity to PRRSV CH-1a strain and HP-PRRSV HuN4 strain in the infected cells, but didn＇t have virus neutralizing activity. Indirect ELISA and virus neutralization results indicated that there was no significant correlation between the anti-B epitope peptide antibodies and NA in pig hyperimmune serums. Base on these findings, we conclude that the B epitope of GP5 protein in HP-PRRSV can not induce NA.

关 键 词：HP-PRRSV GP5 B表位 中和抗体 

分 类 号：S852.65[农业科学—基础兽医学]