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Updated: May 12, 2026

Immuno-fluorescence Assay of Leptospiral Surface-exposed Proteins
Published on: July 1, 2011
[Expression changes of major outer membrane protein antigens in Leptospira interrogans during infection and its
Linli Zheng1, Yumei Ge, Weilin Hu
1Department of Medical Microbiology and Parasitology, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China.
Objective:
To determine expression changes of major outer membrane protein(OMP) antigens of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai during infection of human macrophages and its mechanism.
Methods:
OmpR encoding genes and OmpR-related histidine kinase (HK) encoding gene of L.interrogans strain Lai and their functional domains were predicted using bioinformatics technique. mRNA level changes of the leptospiral major OMP-encoding genes before and after infection of human THP-1 macrophages were detected by real-time fluorescence quantitative RT-PCR. Effects of the OmpR-encoding genes and HK-encoding gene on the expression of leptospiral OMPs during infection were determined by HK-peptide antiserum block assay and closantel inhibitive assays.
Results:
The bioinformatics analysis indicated that LB015 and LB333 were referred to OmpR-encoding genes of the spirochete, while LB014 might act as a OmpR-related HK-encoding gene. After the spirochete infecting THP-1 cells, mRNA levels of leptospiral lipL21, lipL32 and lipL41 genes were rapidly and persistently down-regulated (P <0.01), whereas mRNA levels of leptospiral groEL, mce, loa22 and ligB genes were rapidly but transiently up-regulated (P<0.01). The treatment with closantel and HK-peptide antiserum partly reversed the infection-based down-regulated mRNA levels of lipL21 and lipL48 genes (P <0.01). Moreover, closantel caused a decrease of the infection-based up-regulated mRNA levels of groEL, mce, loa22 and ligB genes (P <0.01).
Conclusion:
Expression levels of L.interrogans strain Lai major OMP antigens present notable changes during infection of human macrophages. There is a group of OmpR-and HK-encoding genes which may play a major role in down-regulation of expression levels of partial OMP antigens during infection.
Insights
Leptospira interrogans outer membrane protein (OMP) antigen expression changes during human macrophage infection were identified. OmpR and histidine kinase (HK) encoding genes significantly influence OMP antigen expression during infection.
Area of Science:
- Microbiology
- Molecular Biology
- Immunology
Background:
- Leptospira interrogans is a pathogenic bacterium causing leptospirosis.
- Outer membrane proteins (OMPs) are crucial for Leptospira pathogenesis and host immune response.
- Understanding OMP expression dynamics during host infection is vital for developing effective treatments.
Purpose of the Study:
- To investigate the expression changes of major OMP antigens of Leptospira interrogans serovar Lai during human macrophage infection.
- To elucidate the underlying molecular mechanisms, focusing on OmpR and histidine kinase (HK) encoding genes.
Main Methods:
- Bioinformatic prediction of OmpR and HK encoding genes in L. interrogans strain Lai.
- Real-time fluorescence quantitative RT-PCR to quantify mRNA levels of OMP-encoding genes before and after THP-1 macrophage infection.
- HK-peptide antiserum block and closantel inhibitive assays to assess the role of OmpR and HK genes in OMP expression.
Main Results:
- Bioinformatics identified LB015 and LB333 as OmpR-encoding genes and LB014 as a potential OmpR-related HK-encoding gene.
- Infection led to rapid and persistent down-regulation of lipL21, lipL32, and lipL41 mRNA, and transient up-regulation of groEL, mce, loa22, and ligB mRNA.
- Closantel and HK-peptide antiserum partially reversed the down-regulation of lipL21 and lipL48, and closantel decreased the up-regulation of other genes.
Conclusions:
- Major OMP antigen expression in L. interrogans strain Lai undergoes significant alterations during human macrophage infection.
- OmpR- and HK-encoding genes play a crucial role in modulating the expression of specific OMP antigens during infection.
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