Heterologous expression of pathogen-specific genes ligA and ligB in the saprophyte Leptospira biflexaconfers enhanced adhesion to cultured cells and fibronectin

BMC Microbiology

Research Article

Henry A Choy¹  David A Haake¹  Mathieu Picardeau²  Cláudio Pereira Figueira³  Mitermayer G Reis³  Albert I Ko⁴  Julio Croda⁵ 

[1] Division of Infectious Diseases, Veterans Affairs Greater Los Angeles Health Care System, 111F, 11301 Wilshire Blvd, 90073, Los Angeles, CACalifornia, USA;Department of Medicine, University of California Los Angeles School of Medicine, Los Angeles, California, USA;Institut Pasteur, Unité de Biologie des Spirochètes, 28 rue du docteur Roux, 75724, Paris Cedex 15, France;Oswaldo Cruz Foundation, Brazilian Ministry of Health, Gonçalo Moniz Research Center, Rua Waldemar Falcão, 121, 40295-001, Salvador, Bahia, Brazil;Oswaldo Cruz Foundation, Brazilian Ministry of Health, Gonçalo Moniz Research Center, Rua Waldemar Falcão, 121, 40295-001, Salvador, Bahia, Brazil;Division of Epidemiology of Microbial Diseases, Department of Epidemiology and Public Health, Yale University School of Medicine, Division of Epidemiology of Microbial Disease, 319 LEPH - 60 College St, 06510, New Haven, CTNew Haven, USA;Oswaldo Cruz Foundation, Brazilian Ministry of Health, Gonçalo Moniz Research Center, Rua Waldemar Falcão, 121, 40295-001, Salvador, Bahia, Brazil;Faculty of Health Sciences, Federal University of Grande Dourados, Brazil;

关键词: Leptospirosis;    Madin Darby Canine Kidney;    Madin Darby Canine Kidney Cell;    Pathogenic Leptospira;    Leptospiral Protein;   

DOI  :  10.1186/1471-2180-11-129

 received in 2011-03-21, accepted in 2011-06-09,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundIn comparison to other bacterial pathogens, our knowledge of the molecular basis of the pathogenesis of leptospirosis is extremely limited. An improved understanding of leptospiral pathogenetic mechanisms requires reliable tools for functional genetic analysis. Leptospiral immunoglobulin-like (Lig) proteins are surface proteins found in pathogenic Leptospira, but not in saprophytes. Here, we describe a system for heterologous expression of the Leptospira interrogans genes ligA and ligB in the saprophyte Leptospira biflexa serovar Patoc.ResultsThe genes encoding LigA and LigB under the control of a constitutive spirochaetal promoter were inserted into the L. biflexa replicative plasmid. We were able to demonstrate expression and surface localization of LigA and LigB in L. biflexa. We found that the expression of the lig genes significantly enhanced the ability of transformed L. biflexa to adhere in vitro to extracellular matrix components and cultured cells, suggesting the involvement of Lig proteins in cell adhesion.ConclusionsThis work reports a complete description of the system we have developed for heterologous expression of pathogen-specific proteins in the saprophytic L. biflexa. We show that expression of LigA and LigB proteins from the pathogen confers a virulence-associated phenotype on L. biflexa, namely adhesion to eukaryotic cells and fibronectin in vitro. This study indicates that L. biflexa can serve as a surrogate host to characterize the role of key virulence factors of the causative agent of leptospirosis.

【 授权许可】

Unknown   
© Figueira et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.