Lefebvre D J, Costers S, Van Doorsselaere J, Misinzo G, Delputte P L, Nauwynck H J
Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Department of Health Care and Biotechnology, KATHO Catholic University College of South-West Flanders, Wilgenstraat 32, 8800 Roeselare, Belgium.
J Gen Virol. 2008 Jan;89(Pt 1):177-187. doi: 10.1099/vir.0.83280-0.
This study examined whether antigenic differences among porcine circovirus type 2 (PCV-2) strains could be detected using monoclonal antibodies (mAbs). A subtractive immunization protocol was used for the genotype 2 post-weaning multisystemic wasting syndrome (PMWS)-associated PCV-2 strain Stoon-1,010. Sixteen stable hybridomas that produced mAbs with an immunoperoxidase monolayer assay (IPMA) titre of 1,000 or more to Stoon-1,010 were obtained. Staining of recombinant PCV-2 virus-like particles demonstrated that all mAbs were directed against the PCV-2 capsid protein. Cross-reactivity of mAbs was tested by IPMA and neutralization assay for genotype 1 strains 48,285, 1,206, VC2,002 and 1,147, and genotype 2 strains 1,121 and 1,103. Eleven mAbs (9C3, 16G12, 21C12, 38C1, 43E10, 55B1, 63H3, 70A7, 94H8, 103H7 and 114C8) recognized all strains in the IPMA and demonstrated neutralization of Stoon-1,010, 48,285, 1,206 and 1,103, but not VC2,002, 1,147 and 1,121. mAbs 31D5, 48B5, 59C6 and 108E8 did not react with genotype 1 strains or had a reduced affinity compared with genotype 2 strains in the IPMA and neutralization assay. mAb 13H4 reacted in the IPMA with PMWS-associated strains Stoon-1,010, 48,285, 1,206 and VC2,002, and the porcine dermatitis and nephropathy syndrome-associated strain 1,147, but not with reproductive failure-associated strains 1,121 and 1,103. mAb 13H4 did not neutralize any of the tested strains. It was concluded that, despite the high amino acid identity of the capsid protein (>or=91 %), antigenic differences at the capsid protein level are present among PCV-2 strains with a different genetic and clinical background.
本研究检测了是否可以使用单克隆抗体(mAb)来检测2型猪圆环病毒(PCV-2)毒株之间的抗原差异。对与2型断奶后多系统消耗综合征(PMWS)相关的PCV-2毒株Stoon-1,010采用了消减免疫方案。获得了16个稳定的杂交瘤,其产生的单克隆抗体通过免疫过氧化物酶单层试验(IPMA)对Stoon-1,010的效价达到1000或更高。重组PCV-2病毒样颗粒的染色表明,所有单克隆抗体均针对PCV-2衣壳蛋白。通过IPMA和中和试验检测了单克隆抗体对1型毒株48,285、1,206、VC2,002和1,147以及2型毒株1,121和1,103的交叉反应性。11种单克隆抗体(9C3、16G12、21C12、38C1、43E10、55B1、63H3、70A7、94H8、103H7和114C8)在IPMA中识别所有毒株,并对Stoon-1,010、48,285、1,206和1,103具有中和作用,但对VC2,002、1,147和1,121没有中和作用。单克隆抗体31D5、48B5、59C6和108E8在IPMA和中和试验中与1型毒株不反应,或与2型毒株相比亲和力降低。单克隆抗体13H4在IPMA中与PMWS相关毒株Stoon-1,010、48,285、1,206和VC2,002以及猪皮炎和肾病综合征相关毒株1,147反应,但与繁殖障碍相关毒株1,121和1,103不反应。单克隆抗体13H4对任何测试毒株均无中和作用。得出的结论是,尽管衣壳蛋白的氨基酸同一性很高(≥91%),但在具有不同遗传和临床背景的PCV-2毒株之间,衣壳蛋白水平存在抗原差异。