问号钩端螺旋体对主要细胞外基质分子黏附作用及其差异性研究

Adhering effect of Leptospira interrongan to major extracellular matrix molecules

目的:了解问号钩端螺旋体(简称钩体)对细胞外基质(ECM)分子的黏附作用及其差异。方法:建立问号钩体黄疸出血群赖型56601株对J774A.1、L929和Vero细胞感染模型,采用Fontana镀银染色法及显微镜检查法,了解问号钩体56601株黏附细胞ECM的效果。建立多种ELISAs,检测问号钩体56601株黏附ECM分子层粘连蛋白(LN)、纤维纤连蛋白(FN)、核心蛋白多糖(DEN)及胶原蛋白1、2、4(COL1,2,4)的作用,采用竞争性抑制试验对上述实验结果进行验证。结果:问号钩体56601株能以一端或两端黏附于L929、J774A.1和Vero细胞的ECM部位。问号钩体56601株可与上述6种ECM分子发生黏附,其中对COL1、LN、COL4的黏附作用较强。问号钩体56601株分别与上述6种ECM分子预孵育后,对相应ECM分子的黏附作用明显下降。结论:ECM是问号钩体黏附宿主细胞的主要部位。LN、FN、DEN、COL1、COL2、COL4均是问号钩体黏附的受体分子,但问号钩体对不同ECM分子的黏附效果有一定差异。

Objective： To determine the adhering ability of Leptospira interrongans to extracellular matrix molecules （ECM） of host cells and its diversity. Methods： The infection models of L. interrongans serogroup lcterohaemorrhagiae serovar lai strain 56601 to J774A. 1, L929 and Vero cells were established. Fontana silver impregnation method was applied to demonstrate the adhering effect of mocrobes to extracellular matrix （ECM） of the cells. ELISA methods were established to detect the adhering effects of L. interrongans to ECM molecules laminin （LN）, fibronectin （FN）, decorin （DEN） and collagenl, 2,4 （COL1,2,4）. A competitive inhibition test was performed to verify the results. Results： L. interrongans strain 56601 adhered the ECMs of L929,J774A. 1 and Vero cells with its one or two sites. L. interrongans strain 56601 adhered all six ECM molecules; the adhering effects to COL1, LN, COL4 were relatively stronger. The adhering effects were markedly decreased after the microbes were pre-incubated with corresponding ECM molecules. Conclusion： L. interrongans adheres to host ceils through ECM molerules ; LN ,FN ,DEN ,COL1 ,COL2 and COL4 are the receptor molecules with different adhesion intensity.