[Membrane location and naturally antibody responses of genus-specific antigen OmpL1s of Leptospira interrogans].
The major aim of this study is to determine the location on outer envelope of the genus-specific antigen OmpL1s of Leptospira interrogans, and the inducement of naturally antibody response and types of the antigen, which will offer the evidences to use OmpL1s as the antigen candidate for developing universal genetic engineering vaccine and detection kit. The serum samples from 156 leptospirosis patients in Sichuan area were detected using microscope agglutination test (MAT). By using PCR plus nucleotide sequence analysis, the genotypes of the dominant L. interrogans serogroups in China were demonstrated. Routine genetic engineering technique was applied to construct the prokaryotic expression systems of genotypes ompL1/1 and ompL1/2, and Ni-NTA affinity chromatography was performed to extract the target recombinant products rOmpL1/1 and rOmpL1/2. Immune aurosol electron microscopy was selected to locate the position of OmpL1s on leptospiral envelope. ELISAs based on rOmpL1s were established to confirm the level and types of specific antibody. The results indicated that L. interrogans serogroup Icterohaemorrhagiae remains to be the most important dominant leptospiral serogroup in Sichuan area. There are two ompL1 genotypes of ompL1/1 and ompL1/2 in the dominant leptospiral serogroup in China. And remarkable differences of the nucleotide and putative amino acid sequence similarities between the two genotypes are present. OmpL1s are the protein molecular that located on the external surface of leptospiral envelope. In the 156 cases of leptospirosis patients' serum samples using different dilutions, the positive rates for rOmpL1/1 or rOmpL1/2 specific IgM are 67.9%-79.5% and 75.0%-75.6%, while for for rOmpL1/1 or rOmpL1/2 specific IgG are 71.8%-79.5% and 75.0%-76.9%, respectively. All the results mentioned above lead to the conclusions that OmpL1s is the leptospiral genus-specific superficial protein antigen of L. interrogans and both rOmpL1/1 and rOmpL1/2 can induce humoral response in individuals naturally infected with L. interrogans as well as produce two serum antibodies IgM and IgG with extensive antigenic-cross reaction. Therefore, rOmpL1/1 and rOmpL1/2 can be used as the antigen candidate for developing universal genetic engineering vaccine and detection kit.
Sun
Bai-li
BL
College of Land and Environment, Shenyang Agriculture Universiy, Shenyang 110161, China. sunbali791012@163.com
Guo
Zong-qi
ZQ
Luo
Dong-jiao
DJ
Yan
Jie
J
D000907:Antibodies, Bacterial / Q000097:blood*; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D007921:Leptospira interrogans / Q000276:immunology*; D016253:Microscopy, Immunoelectron; D016133:Polymerase Chain Reaction
D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; C081904:ompL1 protein, Leptospira