口蹄疫病毒受体猪源整联蛋白αvβ6的细胞定位及组织分布

Cellular Localization and Tissue Profile of Porcine Integrin αvβ6 as FMDV Receptor

利用pcDNA3.1(+)-β6p真核表达载体稳定转染CHOK1细胞,间接免疫荧光法(IFA)检测整联蛋白αvβ6在转染细胞中的表达分布。以间接免疫组化法(IHA)检测整联蛋白αvβ6在健康猪舌、唇、气管等组织的表达分布。采用抗猪源整联蛋白β6亚基的单克隆抗体作为一抗,FITC标记山羊抗小鼠IgG、辣根过氧化物酶标记山羊抗小鼠IgG分别作为二抗。结果表明,pcDNA3.1(+)-β6p真核表达载体稳定转染的CHOK1细胞的细胞膜及细胞质内可见致密的绿色荧光(IFA),猪舌、唇、气管等组织细胞的细胞膜及细胞质内均出现了棕黄色的特异阳性反应物,表明整联蛋白αvβ6受体广泛分布于猪体的器官组织细胞中,为深入研究整联蛋白αvβ6在口蹄疫病毒感染过程中的作用奠定了基础。

CHOK1 cell was stablly transfected by pcDNA3.1（＋）-β6p eucaryon expression vector,then the expressing disposition of integrin αvβ6 was detected by indirect immunofluoresence assay（IFA）.Tissue profile of porcine integrin αvβ6 in porcine tongue,lip,trachea and so on,was detected by indirect immunohistochemistry assay（IHA）.The monoclonal antibody against porcine integrin β6 subunit was used as first antibody,two antibodies of goat against mice IgG labelled with fluorescein isothiocyanate（FITC） and horseradish peroxidase（HRP） respectively were used as secondary antibody.RESULTS： Viridis fluorescence was appeared in cell membrane and cytoplasm of CHOK1 cell stablly transfected by pcDNA3.1（＋）-β6p eucaryon expression vector,buffy idio-positive reaction was appeared in cell membrane and cytoplasm of porcine tongue,lip,trachea and so on,which indicates integrin αvβ6 receptor is generally expressed in porcine tissue cell,which lay a foundation for further understanding of the roles of integrin αvβ6 in FMDV infection.