猪肺炎支原体对猪气管上皮细胞的黏附作用  

Adhesion of Mycoplasma hyopneumoniae to swine tracheal epithelial cells

作　　者：车巧林[1] 余姗姗[1] 刘茂军[1] 甘源[1] 孔猛[1] 邵国青[1] 

机构地区：[1]江苏省农业科学院兽医研究所/农业部兽用生物制品工程技术重点实验室/国家兽用生物制品工程技术研究中心,江苏南京210014

出　　处：《中国兽医学报》2015年第3期388-393,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31100136);中国博士后面上资助项目(2012M510175)

摘　　要：采用气管结扎、链霉蛋白酶灌注冷消化法分离猪气管上皮细胞,并以对数生长期的猪肺炎支原体(Mhp)感染猪气管上皮细胞,通过间接免疫荧光技术定性定量研究不同滴度的猪肺炎支原体对猪气管上皮细胞的黏附及相同滴度的猪肺炎支原体随时间变化对猪气管上皮细胞的黏附。结果表明,分离的猪气管上皮细胞存活率为95%以上,广谱角蛋白染色阳性;Mhp野毒株XLW-1与猪气管上皮细胞37℃作用30min就能看到少量的支原体黏附,作用4h以上时黏附更明显,并且,二者作用1、2、4、6、10h,XLW-1对猪气管上皮细胞的黏附与阴性对照相比差异极显著(P<0.01);1×107、2.5×106 CCU/mL的XLW-1与猪气管上皮细胞37℃作用6h,能看到支原体黏附于细胞表面,并且与阴性对照相比差异极显著(P<0.01),而其他低滴度的XLW-1则看不到黏附。This experiment was conducted to study adhesion of Mycoplasma hyopneumoniae to primary swine tracheal epithelial cells(STEC).The tracheal epithelium was digested with pronase by ligaturing and douching the trachea for overnight.Then STECs were infected with M.hyopneumoniae at exponential growth period.The adhesion of different titer M.hyopneumoniae to STECs was researched qualitatively and quantitatively by indirect immunofluorescence assay.The adhesion to STECs with time variation was also studied in the same way.It was indicated that the survival rate of STECs was over 95%,and pan cytokeratin staining was positive A bits of M.hyopneumoniae XLW-1adhered to STECs at 30 min interaction,and obviously at 4hor more.Differences of adherence to STECs between M.hyopneumoniae XLW-1and KM2 were significant(P<0.01)at 1,2,4,6and 10 h.M.hyopneumoniae XLW-1with a concentration of 1×107 and 2.5×106 CCU/mL could adhere to STECs after interaction for 6hat 37℃,but no adherence at other concentrations.The fluorescent intensity(standing for adherence rate)with XLW-1′s titer of 1×107 and 2.5×106 CCU/mL was significantly higher than other titers and negative control(P<0.01).

关 键 词：猪肺炎支原体 猪气管上皮细胞 黏附 

分 类 号：S858.28[农业科学—临床兽医学]