title: Adhesin activity of Leptospira interrogans lipoprotein identified by in vivo and in vitro shotgun phage display.
abstract: Leptospira interrogans causes leptospirosis, one of the most common zoonotic diseases in the world. This pathogenic spirochete is able to bind to extracellular matrix, to express virulent factors and to cause host death. Until now, there is no effective human vaccine for the disease. Shotgun phage display genomic libraries of L. interrogans were constructed and used for in vivo biopanning in hamsters and screened for ligands able to bind to LLC-PK1 epithelial cells. In both panning procedures, clones coding for the putative lipoprotein LIC12976 were identified and, in order to confirm its adhesin activity, a recombinant protein was produced in Escherichia coli and showed to interact with A31 fibroblasts, LLC-PK1 and Vero epithelial cells in vitro. Moreover, rLIC12976 was shown to bind to laminin, indicating an adhesin function. This protein was also detected in extracts of L. interrogans from different serovars and it was found to be conserved among pathogenic leptospires. Further, the protein was tested as vaccine candidate and immunization of hamsters with LIC12976 did not confer protection against a lethal challenge with the homologous L. interrogans serovar Copenhageni. Nevertheless, LIC12976 seems to act as an adhesin, and may be important for the host-pathogen interaction, so that its study can contribute to the understanding of the virulence mechanisms in pathogenic leptospires.
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, CEP 05503-900 São Paulo, Brazil.
authors.forename: Swiany Silveira
authors.initials: SS
authors.lastname: Lima
authors.forename: Ana Tung Ching
authors.initials: AT
authors.lastname: Ching
authors.forename: Regiane Degan
authors.initials: RD
authors.lastname: Fávaro
authors.forename: Josefa B
authors.initials: JB
authors.lastname: Da Silva
authors.forename: Maria L S
authors.initials: ML
authors.lastname: Oliveira
authors.forename: Eneas
authors.initials: E
authors.lastname: Carvalho
authors.forename: Patrícia A E
authors.initials: PA
authors.lastname: Abreu
authors.forename: Sílvio A
authors.initials: SA
authors.lastname: Vasconcellos
authors.forename: Paulo Lee
authors.initials: PL
authors.lastname: Ho
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics* / Q000502:physiology; D000818:Animals; D002522:Chlorocebus aethiops; D006224:Cricetinae; D054884:Host-Pathogen Interactions; D006801:Humans; D007797:Laminin / Q000378:metabolism; D007921:Leptospira interrogans / Q000235:genetics / Q000472:pathogenicity*; D007922:Leptospirosis / Q000382:microbiology*; D008074:Lipoproteins / Q000235:genetics* / Q000502:physiology; D051379:Mice; D019151:Peptide Library; D014709:Vero Cells
chemical_list: D018829:Adhesins, Bacterial; D007797:Laminin; D008074:Lipoproteins; D019151:Peptide Library