Invivo and Invitro studies on carboxy terminal domains of leptospiral immunoglobulins of L. interrogans Icterohaemorrhagiae

ABSTRACT

Carboxy terminal domains of leptospiral immunoglobulins of rLigA and rLigB were assessed its hemolytic, cytotoxicity effects invitro and its protective efficacy was studied in hamster models. Experimental recombinant proteins of Leptospira interrogans Icterohaemorrhagiae were individually cloned, expressed in prokaryotic system.  Purified expressed proteins confirmed by western blot, assessed for its protective efficacy using hamster models.  100% and 93% protection on homologues challenge was observed with rLigA and rLigB with adjuvant respectively which is evident with increased levels of antibody levels of rLigA and rLigB with 98% and 95% specificity.  Cytotoxicity was maximum with 14% in Daudi cells with with rLigB and 7% and rLigA.  17% and 4% hemolytic activity with rLigA, and rLigB protein and 11% and 10% in pH-7.0 respectively. E. coli (harboring pET15b) membrane fraction used as placebo neither exerted cytotoxicity nor hemolysis was used to compare the leptospiral proteins.  Leptospiral immunoglobulins showed least cytotoxicity and hemolytic activity with increased homologues protection in experimental animals

Keywords: Leptospira recombinant protein, Leptospiral immunoglobulins, immune response, challenge studies, cytotoxic and hemolytic activity.

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