title: Monoclonal antibodies against the leptospiral immunoglobulin-like proteins A and B conserved regions.
abstract: Leptospirosis is an infectious disease caused by pathogenic spirochetes of the genus Leptospira that affects humans and a wide variety of animals. Recently the genomes of Leptospira interrogans, Leptospira borgpetersenii and Leptospira biflexa species were sequenced allowing the identification of new virulence factors involved in survival and pathogenesis of bacteria. LigA and LigB are surface-exposed bacterial adhesins whose expression is correlated with the virulence of Leptospira strains. In this study, we produced and characterized five monoclonal antibodies (MAbs) against a recombinant fragment of LigB (rLigBrep) with approximately 54kDa that comprise the portions of LigA and LigB (domains 2-7). The 5 MAbs obtained were of the IgG1 (2) and IgG2b (3) isotypes and their affinity constants for rLigBrep ranged from 7×10(7) M(-1) to 4×10(8) M(-1). The MAbs were able to react with the native antigen on the L. interrogans, L. borgpetersenii and Leptospira noguchii surfaces by indirect immunofluorescence, immunoblotting and immunoelectron microscopy. These results demonstrate that the MAbs anti-rLigBrep can be useful to complement genetic studies and to aid studies aiming understanding the role of Lig proteins in Leptospira pathogenesis and the development of Lig-based vaccines and improved diagnostic tests for leptospirosis.
authors: Laboratório de Imunologia Aplicada, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, P.O. Box 354, 96010-900, Pelotas, RS, Brazil. leonardogmonte@hotmail.com
authors: Leonardo G
authors: LG
authors: Monte
authors: Fabricio R
authors: FR
authors: Conceição
authors: Mariana L
authors: ML
authors: Coutinho
authors: Fabiana K
authors: FK
authors: Seixas
authors: Everton F
authors: EF
authors: da Silva
authors: Flávia A
authors: FA
authors: Vasconcellos
authors: Luis A S
authors: LA
authors: deCastro
authors: Cláudia P
authors: CP
authors: Hartleben
authors: Odir A
authors: OA
authors: Dellagostin
authors: José A G
authors: JA
authors: Aleixo
mesh_terms: D018829:Adhesins, Bacterial / Q000276:immunology; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D058846:Antibodies, Monoclonal, Murine-Derived / Q000276:immunology* / Q000302:isolation & purification; D000942:Antigens, Bacterial / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology; D002846:Chromatography, Affinity; D018604:Epitope Mapping; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006825:Hybridomas / Q000276:immunology*; D015151:Immunoblotting; D007132:Immunoglobulin Isotypes / Q000276:immunology; D007921:Leptospira interrogans / Q000276:immunology* / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D046529:Microscopy, Electron, Transmission; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D058846:Antibodies, Monoclonal, Murine-Derived; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D007132:Immunoglobulin Isotypes; C487005:LigA-m protein, Leptospira interrogans; C487006:LigB-m protein, Leptospira interrogans; D011994:Recombinant Proteins