Leptospira Hemolysin Biological Functions And Activity Of Lipase Hydrolysis Explore

Pathogenic genome sequencing of Leptospira interrogans has been completed by Chinese National Human Genome Center at Shanghai, China. The ORFs scanning results of Leptospira interrogans serogroup Icterohaemorrhagiae serovar type strain (Lai) show that there are eleven genes which show high similarity to hemolysin genes reported in NCBI/Genbank database. The amino acid sequences encoded by these genes also show high similarity to hemolysins reported in SwissProt/TrEMBL database. Eight of them were cloned, expressed and their hemolytic activities were studied. In this work, hemolytic and cytotoxic activities of Leptospira interrogans hemolysin LA3937 and LA1029 were studied further because of their marked hemolytic activity from different levels, such as cellular level, subcellular level, molecular level, histological level.Ⅰ .Cellular and subcellular level : BALB/C mouse lymphocytes and macrophages, human liver L-02 cells and human kidney 293 cells were exposed to hemolysins and changes of cells were studied through convert microscopy, transmission electron microscopy, scanning electron microscopy and fluorescence confocal microscopy. Ⅱ .Molecular level : A. Toxins are considered to be principal virulence factors contributing to many inflammatory diseases in humans and animals. Sub-lytic concentrations of toxins cause a cellular activation to generate various proinflammatory mediators such as cytokines. B. Apoptosis is the process of programmed cell death or cell suicide, which can be detected by flow cytometry . C. Microarray assay analysis : Hemolysin LA3937 was added into human liver L-02 cells at two concentrations, lower (10ug/ml) and higher (30ug/ml). We screened gene expressin by microarray consisting of 13824 focused genes to examine the holistic gene expression. Ⅲ .Histological level : Hemolysin LA3937 was injected into KM mouse through intravenous injection, and the histological changes of lung, liver...