不同毒力猪肺炎支原体侵入宿主细胞的观察  

Invasion of Host Cells by Different Virulent Mycoplasma hyopneumoniae

作　　者：车巧林[1] 刘蓓蓓[1] 刘茂军[1] 熊祺琰[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,南京210014

出　　处：《畜牧兽医学报》2014年第6期981-988,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

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

摘　　要：旨在研究不同毒力猪肺炎支原体对宿主细胞的侵入以及侵入后在宿主细胞内的分布位置。首先用猪肺炎支原体野毒株XLW-1感染PK15细胞不同时间后,间接免疫荧光染色,激光扫描共聚焦显微镜对PK15细胞进行断层扫描,以观察XLW-1能否侵入细胞以及感染时间对XLW-1侵入细胞的影响。然后,在能侵入细胞的基础上,用不同毒力的猪肺炎支原体感染宿主细胞STEC、PK15、SJPL和3D4/21,观察它们对宿主细胞的侵入情况。结果表明,猪肺炎支原体野毒株XLW-1与PK15细胞作用8h时不能侵入PK15,作用10h以上时可以侵入PK15,并且随着作用时间的延长侵入的猪肺炎支原体也在增加。研究还表明猪肺炎支原体野毒株XLW-1、168致弱株和168疫苗株均能侵入STEC、PK15、SJPL和3D4/21。并且,STEC中3个菌株均随机分布在细胞质中;PK15中3个菌株均主要分布在靠近细胞膜和细胞两端狭长的细胞质中;SJPL中XLW-1和168疫苗株主要分布在贴近细胞膜内侧的细胞质中,168致弱株则散布在细胞质中;3D4/21中XLW-1和168致弱株主要分布在贴近细胞膜内侧的细胞质中,168疫苗株则在整个细胞质中分布较多。结果提示不同毒力的猪肺炎支原体均能侵入宿主细胞STEC、PK15、SJPL和3D4/21。This experiment was conducted to study the invasion and location in host cells of different virulent Mycoplasma hyopneumoniae.The ability of wild strain XLW-1to invade PK15cells at different infection time interval was investigated firstly by using indirect immunofluorescence technique and laser scanning confocal microscope.Then,the invasion to swine tracheal epithelial cells(STEC),PK15,SJPL and 3D4/21of different virulent M.hyopneumoniae strains was observed.The results showed that 1)M.hyopneumoniae wild strains XLW-1can intrude into PK15cells up to 10hpost infection and no internalization was observed before 8h;2)The amount of intracellular M.hyopneumoniae XLW-1was increased as infection time;3)M.hyopneumoniae wild strainsXLW-1,168attenuated strains and vaccine strains all could invade STEC,PK15,SJPL and 3D4/21cells;4)The ingested wild strainsXLW-1,168attenuated strains and vaccine strains were randomly distributed in the cytoplasm of STEC,and mainly distributed in the cytoplasm oflong and narrow two polars and cytoplasm near cytomembrane in PK15;The internalized wild strainsXLW-1and 168vaccine strains principally concentrated in the cytoplasm close to medial surface of SJPL cells membrane,but 168attenuated strains randomly interspersed in the cytoplasm of SJPL cells;The incursive wild strainsXLW-1and 168attenuated strains principally distributed in the cytoplasm close to inboard surface of 3D4/21cytomembrane,and 168vaccine strains randomly spreaded in the cytoplasm of 3D4/21cells.These results demonstated that M.hyopneumoniae of different virulence all can invade host cells STEC,PK15,SJPL and 3D4/21.

关 键 词：猪肺炎支原体 不同毒力 宿主细胞 侵入 

分 类 号：S852.62[农业科学—基础兽医学]