Determination of the genus-specific antigens in outer membrane proteins from the strains ofLeptospira interrogans andLeptospira biflexa with different virulence

Objective: To determine the existence of genus-specific antigens in outer membrane proteins (OMPs) ofleptospira with different virulence. Methods: Microscope agglutination test (MAT) was applied to detect the agglutination between commercial rabbit antiserum against leptospiral genus-specific TR/Patoc I antigen and 17 strains ofLeptospira interrongans belonging to 15 serogroups and 2 strains ofLeptospira biflexa belonging to 2 serogroups. The outer envelopes (OEs) ofLinterrogans serogroupIcterohaemorrhagiae serovarlai strainlai (56601) with strong virulence and serogroupPomona serovarpomona strainLuo (56608) with low virulence, andL. biflexa serogroupSemaranga serovarpatoc strainPatoc I without virulence were prepared by using the method reported in Auranet al. (1972). OMPs in the OEs were obtained by treatment with sodium deoxycholate. SDS-PAGE and western blot were used for analyzing the features of the OMPs on electrophoretic pattern and the immunoreactivity to the antiserum against TR/Patoc I antigen, respectively. Results: All the tested strains belonging to different leptospiral serogroups agglutinated to the antiserum against leptospiral genus-specific TR/Patoc I antigen with agglutination titers ranging from 1:256–1:512. A similar SDS-PAGE pattern of the OMPs from the three strains ofleptospira with different virulence was shown and the molecular weight of a major protein fragment in the OMPs was found to be approximately 60 Kda. A positive protein fragment with approximately 32 KDa confirmed by Western blot, was able to react with the antiserum against leptospiral genus-specific TR/Patoc I antigen, and was found in each the OMPs of the three stains ofleptospira. Conclusion: There are genus-specific antigens on the surface ofL.interrogans andL.biflexa. The OMP with molecular weight of 32 KDa may be one of the genus-specific protein antigens ofleptospira. Project (No. 39970678) supported by the National Natural Science Foundation of China