Effect And Diversity Of Leptospira Interrongan Adhering Host Cells And Major Extracellular Matrix Molecules

Objective: To determine the ability and diversity of Leptospira interrongans adhering to extracellular matrix (ECM) of host cells, and to determine the distribution and content of diferent extracellular matrix (ECM) molecules in various host cells of Leptospira interrongan.Methods Models of L.interrongans serogroup Icterohaemorrhagiae serovar lai strain 56601 infecting J774A.1, L929 and Vero cells were established, and Fontana silver impregnation method plus microscopy were applied to demonstrate the effect of L.interrongans strain 56601 adhere extracellular matrix (ECM) of the cells. ELISAs were established to detect the effects of L.interrongans strain 56601 adhere 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 of adhesion mentioned above. Immunity fluorescence dyeing method and confocal laser microscopy were used to observe the distribution of FN, LN, DEN and COL1～4 in THP-1, J774A.1, HEK293, NRK, COS-7, NRK-49F, L-02, IAR-20 and HUVEC cells. Different kinds of ELISAs were established to verify the results and detect the content of these ECM molecules in different cells.Results L.interrongans strain 56601 could use its one or two bodies to adhere the positions where were the ECMs of L929, J774A.1 and Vero cells. L.interrongans strain 56601 could adhere each the six ECM molecules in which the adhering effects to COL1, LN, COL4 were relatively stronger. The effects of L.interrongans strain 56601 adhering to the six ECM molecules were remarkably decreased after the microbe was pre-incubated with each the molecules. The distribution and content of different ECM molecules in host cells are various. THP-1 and J774A.1 cells contain FN only. HEK293 cells cotain FN and COL4. NRK contain FN and LN, the content of FN is high.. COS-7 contain LN, COL3 and COL4, the content of LN and COL4 is high. NRK-49F contain FN, LN, DEN, COO and COL4, the content of FN, DEN and COL4 is high. L-02 contain FN, LN, COL3 and COL4, the content of FN and COL4 is high. IAR-2 contain COL4, HUVEC contain FN, LN, COL3 and COL4, the content of FN and COL4 is high.Conclusion ECM is the position of L.interrongans adhering to host cells. LN, FN, DEN, COL1, COL2, COL 4 are the receptor molecules of adhesion secreted by L.interrongans, but the different ECM molecules show different adhered effects. The distribution of different ECM molecules in host cells of Leptospira interrongan is quite different, and the content is also distinguishing.