不同毒力猪肺炎支原体侵入宿主细胞的观察

doi:10.11843/j.issn.0366-6964.2014.06.017

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (6): 981-988.doi: 10.11843/j.issn.0366-6964.2014.06.017

不同毒力猪肺炎支原体侵入宿主细胞的观察

车巧林，刘蓓蓓，刘茂军，熊祺琰，邵国青^*

（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）

收稿日期:2013-10-14 出版日期:2014-06-23 发布日期:2014-06-23
通讯作者: 邵国青，研究员，E-mail：gqshaojaas@gmail.com
作者简介:车巧林（1982-），女，山西吕梁人，助理研究员，博士，主要从事猪肺炎支原体致病机理研究，E-mail：cheqiaolin@163.com
基金资助:
国家自然科学基金（31100136）；中国博士后面上项目（2012M510175）

Invasion of Host Cells by Different Virulent Mycoplasma hyopneumoniae

CHE Qiao-lin，LIU Bei-bei，LIU Mao-jun，XIONG Qi-yan，SHAO Guo-qing^*

(Institute of Veterinary Medicine，Jiangsu Academy of Agricultural Sciences，Key Laboratory of Veterinary Biological Engineering and Technology of Ministry of Agriculture，National Center for Engineering Research of Veterinary Bio-products，Nanjing 210014 ， China)

Received:2013-10-14 Online:2014-06-23 Published:2014-06-23

摘要/Abstract

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8 h时不能侵入PK15，作用10 h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。

关键词: 猪肺炎支原体, 不同毒力, 宿主细胞, 侵入

Abstract:

This experiment was conducted to study the invasion and location in host cells of different virulent Mycoplasma hyopneumoniae.The ability of wild strain XLW-1 to invade PK15 cells 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 3D4/21 of different virulent M.hyopneumoniae strains was observed.The results showed that 1) M.hyopneumoniae wild strains XLW-1 can intrude into PK15 cells up to 10 h post infection and no internalization was observed before 8 h; 2) The amount of intracellular M.hyopneumoniae XLW-1 was increased as infection time; 3) M.hyopneumoniae wild strainsXLW-1，168 attenuated strains and vaccine strains all could invade STEC，PK15，SJPL and 3D4/21 cells; 4) The ingested wild strainsXLW-1，168 attenuated strains and vaccine strains were randomly distributed in the cytoplasm of STEC，and mainly distributed in the cytoplasm of long and narrow two polars and cytoplasm near cytomembrane in PK15; The internalized wild strainsXLW-1 and 168 vaccine strains principally concentrated in the cytoplasm close to medial surface of SJPL cells membrane，but 168 attenuated strains randomly interspersed in the cytoplasm of SJPL cells; The incursive wild strainsXLW-1 and 168 attenuated strains principally distributed in the cytoplasm close to inboard surface of 3D4/21 cytomembrane，and 168 vaccine strains randomly spreaded in the cytoplasm of 3D4/21 cells.These results demonstated that M.hyopneumoniae of different virulence all can invade host cells STEC，PK15，SJPL and 3D4/21.

Key words: Mycoplasma hyopneumoniae, different virulence, host cells, invasion

中图分类号:

S852.62

车巧林，刘蓓蓓，刘茂军，熊祺琰，邵国青. 不同毒力猪肺炎支原体侵入宿主细胞的观察[J]. 畜牧兽医学报, 2014, 45(6): 981-988.

CHE Qiao-lin，LIU Bei-bei，LIU Mao-jun，XIONG Qi-yan，SHAO Guo-qing. Invasion of Host Cells by Different Virulent Mycoplasma hyopneumoniae[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(6): 981-988.

参考文献

［1］LO S C，DAWSON M S，WONG D M，et al.Identification of Mycoplasma incognitus infection in patients with AIDS：an immunohistochemical，in situ hybridization and ultrastructural study ［J］.Am J Trop Med Hyg，1989，41(5):601-616.
［2］STADTLÄNDER C T，WATSON H L，SIMECKA J W，et al.Cytopathogenicity of Mycoplasma fermentans (including strain incognitus) ［J］.Clin Infect Dis，1993，17 (Suppl 1):S289-S301.
［3］TAYLOR-ROBINSON D，DAVIES H A，SARATHCHANDRA P，et al.Intracellular location of mycoplasmas in cultured cells demonstrated by immunocytochemistry and electron microscopy ［J］.Int J Exp Pathol，1991，72(6):705-714.
［4］KORNSPAN J D，TARSHIS M，ROTTEM S.Invasion of melanoma cells by Mycoplasma hyorhinis：enhancement by protease treatment ［J］.Infect Immun，2010，78(2):611-617.
［5］YAVLOVICH A，KATZENELL A，TARSHIS M，et al.Mycoplasma fermentans binds to and invades HeLa cells：involvement of plasminogen and urokinase ［J］.Infect Immun，2004，72(9):5004-5011.
［6］BOROVSKY Z，TARSHIS M，ZHANG P，et al.Protein kinase C activation and vacuolation in HeLa cells invaded by Mycoplasma penetrans ［J］.J Med Microbiol，1998，47(10):915-922.
［7］VOGL G，PLAICKNER A，SZATHMARY S，et al.Mycoplasma gallisepticum invades chicken erythrocytes during infection ［J］.Infect Immun，2008，76(1):71-77.
［8］LO S C，HAYES M M，KOTANI H，et al.Adhesion onto and invasion into mammalian cells by Mycoplasma penetrans：a newly isolated mycoplasma from patients with AIDS［J］.Mod Pathol，1993，6(3)：276-280.
［9］ANDREEV J，BOROVSKY Z，ROSENSHINE I，et al.Invasion of HeLa cells by Mycoplasma penetrans and the induction of tyrosine phosphorylation of a 145 kDa host cell protein ［J］.FEMS Microbiol Lett，1995,132(3)：189-194.
［10］YAVLOVICH A，TARSHIS M，ROTTEM S.Internalization and intracellular survival of Mycoplasma pneumoniae by nonphagocytic cells ［J］.FEMS Microbiol Lett，2004，233(2):241-246.
［11］JENSEN J G，BLOM J，LIND K.Intracellular location of Mycoplasma genitalium in cultured vero cells as demonstrated by electron microscopy ［J］.Int J Exp Pathol,1994，75(2)：91-98.
［12］WINNER F，ROSENGARTEN R，CITTI C.In vitro cell invasion of Mycoplasma gallisepticum ［J］. Infect Immun，2000，68(7):4238-4244.
［13］CITTI C，BROWNING G F，ROSENGARTEN R.Phenotypic diversity and cell invasion in host subversion by pathogenic mycoplasmas［M］//Blanchard A，Browning GF.Mycoplasmas：pathogenesis，molecular biology，and emerging strategies for control.Wymondham，Norfolk，United Kingdom：Horizon Bioscience，2005：439-483.
［14］DAXBOECK F，KHANAKAH G，BAUER C，et al.Detection of Mycoplasma pneumoniae in serum specimens from patients with mycoplasma pneumonia by PCR ［J］.Int J Med Microbiol，2005，295(4):279-285.
［15］MUCH P，WINNER F，STIPKOVITS L，et al.Mycoplasma gallisepticum：influence of cell invasiveness on the outcome of experimental infection in chickens ［J］.FEMS Immunol Med Microbiol，2002，34(3):181-186.
［16］ORTIZ DE SOLÓRZANO C，GARCÍA R E，JONES A，et al.Segmentation of confocal microscope images of cell nuclei in thick tissue sections ［J］ .J Microsc，1999,193(Pt 3):212-226.
［17］IBÁÑEZ-LÓPEZ C，ESCOBAR I，SAAVEDRA G，et al.Optical-sectioning improvement in two-color excitation scanning microscopy［J］.Microsc Res Tech，2004，64(2):96-102.
［18］车巧林.猪肺炎支原体体外感染模型建立及强弱毒株感染差异研究［D］.南京：江苏省农业科学院，2013.

[1]	刘璐, 郭杰, 朱可蒙, 王宁, 辛九庆, 刘玉芬, 刘恒贵. 利用酵母随机展示技术对猪肺炎支原体P97上B细胞表位的筛选和鉴定[J]. 畜牧兽医学报, 2018, 49(7): 1492-1499.
[2]	刘璐, 郭杰, 朱可蒙, 王宁, 辛九庆, 刘玉芬, 刘恒贵. 利用酵母随机展示技术对猪肺炎支原体P97上B细胞表位的筛选和鉴定[J]. 畜牧兽医学报, 2018, 49(7): 1492-1499.
[3]	冯艳艳, 王海燕, 刘蓓蓓, 韦艳娜, 张珍珍, 白昀, 倪博, 邵国青, 雷治海, 冯志新. 猪肺炎支原体不同毒力菌株对猪气管上皮3D细胞的损伤差异与机制分析[J]. 畜牧兽医学报, 2018, 49(2): 378-387.
[4]	冯艳艳, 王海燕, 刘蓓蓓, 韦艳娜, 张珍珍, 白昀, 倪博, 邵国青, 雷治海, 冯志新. 猪肺炎支原体不同毒力菌株对猪气管上皮3D细胞的损伤差异与机制分析[J]. 畜牧兽医学报, 2018, 49(2): 378-387.
[5]	武昱孜, 熊祺琰, 刘蓓蓓, 张珍珍, 王佳, 华利忠, 韦艳娜, 邵国青. 猪肺炎支原体和猪鼻支原体双重荧光定量PCR检测方法的建立[J]. 畜牧兽医学报, 2017, 48(8): 1491-1498.
[6]	武昱孜, 熊祺琰, 刘蓓蓓, 张珍珍, 王佳, 华利忠, 韦艳娜, 邵国青. 猪肺炎支原体和猪鼻支原体双重荧光定量PCR检测方法的建立[J]. 畜牧兽医学报, 2017, 48(8): 1491-1498.
[7]	武昱孜, 邢宪平, 韦艳娜, 熊祺琰, 冯志新, 邵国青. 不同猪支原体肺炎活疫苗菌株P46和P97基因序列差异性分析[J]. 畜牧兽医学报, 2017, 48(12): 2365-2373.
[8]	武昱孜, 邢宪平, 韦艳娜, 熊祺琰, 冯志新, 邵国青. 不同猪支原体肺炎活疫苗菌株P46和P97基因序列差异性分析[J]. 畜牧兽医学报, 2017, 48(12): 2365-2373.
[9]	吴彩艳，王祯，李娟，林栩慧，廖申权，戚南山，吕敏娜，孙铭飞. 宿主细胞F-actin聚集在顶复门原虫入侵过程中的作用研究[J]. 畜牧兽医学报, 2016, 47(3): 423-428.
[10]	吴彩艳，王祯，李娟，林栩慧，廖申权，戚南山，吕敏娜，孙铭飞. 宿主细胞F-actin聚集在顶复门原虫入侵过程中的作用研究[J]. 畜牧兽医学报, 2016, 47(3): 423-428.
[11]	张妍，宋纪伟，曾范利，时坤，李健明，刘杨，刘菲，孙凡婷，杜锐. 不同毒力结核分枝杆菌对THP-1细胞凋亡及细胞因子表达的影响[J]. 畜牧兽医学报, 2015, 46(9): 1600-1605.
[12]	张妍，宋纪伟，曾范利，时坤，李健明，刘杨，刘菲，孙凡婷，杜锐. 不同毒力结核分枝杆菌对THP-1细胞凋亡及细胞因子表达的影响[J]. 畜牧兽医学报, 2015, 46(9): 1600-1605.
[13]	李彦伟，刘茂军，刘蓓蓓，武昱孜，倪博，白方方，纪燕，张映，邵国青. 猪肺炎支原体对上皮细胞氧化损伤的分析[J]. 畜牧兽医学报, 2014, 45(7): 1202-1206.
[14]	李彦伟，刘茂军，刘蓓蓓，武昱孜，倪博，白方方，纪燕，张映，邵国青. 猪肺炎支原体对上皮细胞氧化损伤的分析[J]. 畜牧兽医学报, 2014, 45(7): 1202-1206.
[15]	车巧林，刘蓓蓓，刘茂军，熊祺琰，邵国青. 不同毒力猪肺炎支原体侵入宿主细胞的观察[J]. 畜牧兽医学报, 2014, 45(6): 981-988.

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

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价