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Immuno-fluorescence Assay of Leptospiral Surface-exposed Proteins
Published on: July 1, 2011
The OmpL37 surface-exposed protein is expressed by pathogenic Leptospira during infection and binds skin and vascular
Marija Pinne1, Henry A Choy, David A Haake
1Research Service, 151, Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles, California, USA. mpinne@ucla.edu
Abstract:
Pathogenic Leptospira spp. shed in the urine of reservoir hosts into freshwater can be transmitted to a susceptible host through skin abrasions or mucous membranes causing leptospirosis. The infection process involves the ability of leptospires to adhere to cell surface and extracellular matrix components, a crucial step for dissemination and colonization of host tissues. Therefore, the elucidation of novel mediators of host-pathogen interaction is important in the discovery of virulence factors involved in the pathogenesis of leptospirosis. In this study, we assess the functional roles of transmembrane outer membrane proteins OmpL36 (LIC13166), OmpL37 (LIC12263), and OmpL47 (LIC13050), which we recently identified on the leptospiral surface. We determine the capacity of these proteins to bind to host tissue components by enzyme-linked immunosorbent assay. OmpL37 binds elastin preferentially, exhibiting dose-dependent, saturating binding to human skin (K(d), 104±19 nM) and aortic elastin (K(d), 152±27 nM). It also binds fibrinogen (K(d), 244±15 nM), fibrinogen fragment D (K(d), 132±30 nM), plasma fibronectin (K(d), 359±68 nM), and murine laminin (K(d), 410±81 nM). The binding to human skin elastin by both recombinant OmpL37 and live Leptospira interrogans is specifically enhanced by rabbit antiserum for OmpL37, suggesting the involvement of OmpL37 in leptospiral binding to elastin and also the possibility that host-generated antibodies may promote rather than inhibit the adherence of leptospires to elastin-rich tissues. Further, we demonstrate that OmpL37 is recognized by acute and convalescent leptospirosis patient sera and also by Leptospira-infected hamster sera. Finally, OmpL37 protein is detected in pathogenic Leptospira serovars and not in saprophytic Leptospira. Thus, OmpL37 is a novel elastin-binding protein of pathogenic Leptospira that may be promoting attachment of Leptospira to host tissues.
Insights
Pathogenic Leptospira outer membrane protein OmpL37 binds host tissue components like elastin, aiding bacterial attachment and leptospirosis pathogenesis. This protein is recognized by patient sera and found only in pathogenic Leptospira strains.
Area of Science:
- Microbiology
- Immunology
- Pathogenesis
Background:
- Leptospirosis is caused by pathogenic Leptospira spp. transmitted through contaminated water.
- Bacterial adherence to host tissues is crucial for leptospiral dissemination and colonization.
- Identifying host-pathogen interaction mediators is key to understanding leptospirosis pathogenesis.
Purpose of the Study:
- To investigate the functional roles of novel leptospiral outer membrane proteins (OmpL36, OmpL37, OmpL47) in host-pathogen interactions.
- To determine the binding capabilities of these proteins to host tissue components.
- To assess the potential role of OmpL37 in the pathogenesis of leptospirosis.
Main Methods:
- Enzyme-linked immunosorbent assay (ELISA) to assess protein binding to host matrix components.
- Characterization of binding kinetics (K(d) values) for OmpL37 with various host ligands.
- Detection of OmpL37 in pathogenic Leptospira serovars and its recognition by patient and animal sera.
Main Results:
- OmpL37 demonstrated preferential and dose-dependent binding to elastin, fibrinogen, fibronectin, and laminin.
- OmpL37 binding to human skin elastin was enhanced by specific antiserum, suggesting a role in adherence.
- OmpL37 was recognized by sera from leptospirosis patients and infected hamsters, and detected in pathogenic Leptospira strains.
Conclusions:
- OmpL37 is a novel elastin-binding protein expressed by pathogenic Leptospira.
- OmpL37 likely contributes to the attachment of Leptospira to elastin-rich host tissues, playing a role in leptospirosis pathogenesis.
- OmpL37 represents a potential target for diagnostic and therapeutic strategies against leptospirosis.
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