Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/7498
Title: Heterologous expression of pathogen-specific genes ligA and ligB in the saprophyte Leptospira biflexa confers enhanced adhesion to cultured cells and fibronectin.
Authors: Figueira, Cláudio Pereira
Croda, Julio Henrique Rosa
Choy, Henry A
Haake, David A
Reis, Mitermayer Galvão dos
Ko, Albert Icksang
Picardeau, Mathieu
Affilliation: Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil
Veterans Affairs Greater Los Angeles Health Care System. Division of Infectious Diseases. California, USA / University of California Los Angeles School of Medicine. Department of Medicine. Los Angeles, California, USA
Veterans Affairs Greater Los Angeles Health Care System. Division of Infectious Diseases. California, USA / University of California Los Angeles School of Medicine. Department of Medicine. Los Angeles, California, USA
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil / University of California Los Angeles School of Medicine. Department of Medicine. Los Angeles, California, USA
Institut Pasteur. Unité de Biologie des Spirochètes. Paris, France
Abstract: BACKGROUND: In 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. RESULTS: The 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. CONCLUSIONS: This 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.
DeCS: Antígenos de Bactérias/metabolismo
Aderência Bacteriana
Células Epiteliais/microbiologia
Fibronectinas/metabolismo
Leptospira/patogenicidade
Fatores de Virulência/metabolismo
Animais
Antígenos de Bactérias/genética
Linhagem Celular
Cães
Expressão Gênica
Vetores Genéticos
Leptospira/genética
Plasmídeos
Proteínas Recombinantes/genética
Proteínas Recombinantes/metabolismo
Fatores de Virulência/genética
Issue Date: 2011
Publisher: Public Library of Science
Citation: FIGUEIRA, C. et al. Heterologous expression of pathogen-specific genes ligA and ligB in the saprophyte Leptospira biflexa confers enhanced adhesion to cultured cells and fibronectin. BMC Microbiology, v. 11, p. 129, 2011.
ISSN: 1471-2180
Copyright: open access
Appears in Collections:BA - IGM - Artigos de Periódicos

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