鸭源鸡杆菌对HeLa细胞的黏附和侵袭特性

Adhesion and Invasion Characteristics of Gallibacterium anatis to HeLa Cells

为研究鸭源鸡杆菌(G.anatis)体外黏附和侵袭细胞的能力,选择HeLa细胞为感染模型,分别以细菌黏附和侵袭HeLa细胞数量为指标,借助革兰和姬姆萨染色及电镜观察来研究2株不同来源的鸭源鸡杆菌对HeLa细胞的黏附和侵袭特性。结果显示:黏附计数表明鸭源鸡杆菌Yu-PDS-RZ-1-SLG能高水平黏附HeLa细胞,菌株感染30、60、90、120、150min后每细胞黏附的细菌数分别为0.84、4.68、8.52、9.27、8.2个,而菌株F149T对HeLa细胞有较低的黏附作用。侵袭试验表明Yu-PDS-RZ-1-SLG有低度侵袭力,侵袭的细菌数在150min时达到最大,约8.27·1 000细胞-1,随后细胞破碎,细胞逐渐脱落,而菌株F149T未检测到侵袭力。染色及电镜试验进一步证实鸭源鸡杆菌Yu-PDS-RZ-1-SLG可黏附于HeLa细胞表面并侵入细胞内部。菌株经胰蛋白酶处理后,细菌的黏附率显著降低,表明细菌的表面蛋白参与黏附。黏附和入侵特性是鸭源鸡杆菌定殖在组织表面重要的能力,该细胞模型的建立为进一步研究其致病机制提供了条件。

In order to determine the cell adhesion and invasion characteristics of Gallibacterium anatis(G.anatis),an in vitro HeLa cell model was established.The adhesion and invasion characteristics of 2strains were evaluated by means of adhesion and invasion experiment,Gram and Giemsa staining,Scanning electron microscopy and Transmission electron microscopy.Adhesion counting assay demonstrated that the high adhesive of G.anatis Yu-PDS-RZ-1-SLG strain and adhering bacteria numbers were 0.84·cell-1,4.68·cell-1,8.52·cell-1,9.27·cell-1 and8.2 ·cell-1 at 30,60,90,120,150 min after infection,respectively.G.anatis F149 Tstrain revealed a weak adhesive level.Invasion assay showed weak invasive of G.anatis Yu-PDS-RZ-1-SLG strain and no invasion of G.anatis F149 Tstrain,and invading bacteria numbers of G.anatis Yu-PDS-RZ-1-SLG strain reached the maximum at 150 min,about 8.27 per 1 000 cells,but cell disruption,cell activity weakened subsequently.Staining results and electron microscopy experiments further confirmed that G.anatis Yu-PDS-RZ-1-SLG strain adhere to surface and invade Hela cells.Adherence was prevented by treating bacterial cells with trypsin,suggesting the participation of proteins in this process.The adhesive and invasive capacity could be an important ability for colonization of tissue surfaces of G.anatis.The establishment of cell model provided the basis for further research on pathogenic mechanism of G.anatis.