title: Leptospira interrogans Lsa23 protein recruits plasminogen, factor H and C4BP from normal human serum and mediates C3b and C4b degradation.
abstract: It has been reported that pathogenic Leptospira are resistant to normal human serum (NHS) due to their ability to evade the complement immune system by interacting with factor H (FH) and C4b-binding protein (C4BP) regulators. Moreover, plasmin generation on the leptospiral surface diminishes C3b and IgG deposition, decreasing opsonophagocytosis by immune competent cells. We have previously reported that Lsa23 (LIC11360) is a multipurpose protein capable of binding purified extracellular matrix molecules, FH, C4BP and plasminogen (PLG)/plasmin in the presence of PLG activators. In this work, we provide further evidence that Lsa23 is located at the bacterial surface by using immunofluorescence microscopy. We show that Lsa23 has the ability to acquire FH, C4BP and PLG from NHS, and use these interactions to evade innate immunity. The binding with the complement regulators FH and C4BP preserves factor I (FI) activity, leading to C3b and C4b degradation products, respectively. C3b and C4b alpha-chain cleavage was also observed when Lsa23 bound to PLG generating plasmin, an effect blocked by the protease inhibitor aprotinin. Lsa23 also inhibited lytic activity by NHS mediated by both classical and alternative complement pathways. Thus, Lsa23 has the ability to block both pathways of the complement system, and may help pathogenic Leptospira to escape complement-mediated clearance in human hosts. Indeed, NHS treated with Lsa23 confers a partial serum resistance phenotype to Leptospira biflexa, whereas blocking this protein with anti-Lsa23 renders pathogenic L. interrogans more susceptible to complement-mediated killing. Thus, Lsa23 is a multifunctional protein involved in many pathways, featuring C4b cleavage by plasmin, knowledge that may help in the development of preventive approaches to intervene with human complement escape by this versatile pathogen.
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil.|Instituto de Ciências Biomédicas, Universidade de São Paulo, Avenida Professor Lineu Prestes, 1730, 05508-900, São Paulo, SP, Brazil.
authors.forename: Gabriela H
authors.initials: GH
authors.lastname: Siqueira
authors.affiliation: Instituto Adolfo Lutz, Avenida Doutor Arnaldo, 355, 01246-000, São Paulo, SP, Brazil.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Atzingen
authors.affiliation: Laboratório de Zoonoses Bacterianas do VPS, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Avenida Professor Dr Orlando Marques de Paiva, 87, 05508-270, São Paulo, SP, Brazil.
authors.forename: Gisele O
authors.initials: GO
authors.lastname: de Souza
authors.affiliation: Laboratório de Zoonoses Bacterianas do VPS, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Avenida Professor Dr Orlando Marques de Paiva, 87, 05508-270, São Paulo, SP, Brazil.
authors.forename: Silvio A
authors.initials: SA
authors.lastname: Vasconcellos
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil.|Instituto de Ciências Biomédicas, Universidade de São Paulo, Avenida Professor Lineu Prestes, 1730, 05508-900, São Paulo, SP, Brazil.
authors.forename: Ana L T O
authors.initials: ALTO
authors.lastname: Nascimento
mesh_terms: D001426:Bacterial Proteins / Q000276:immunology*; D003179:Complement C3b / Q000378:metabolism*; D015935:Complement C4b / Q000378:metabolism*; D050716:Complement C4b-Binding Protein / Q000378:metabolism*; D017242:Complement Factor H / Q000378:metabolism*; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D007921:Leptospira interrogans / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D008565:Membrane Proteins / Q000276:immunology*; D008856:Microscopy, Fluorescence; D010587:Phagocytosis / Q000276:immunology; D010958:Plasminogen / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D050716:Complement C4b-Binding Protein; D008565:Membrane Proteins; D003179:Complement C3b; D015935:Complement C4b; D017242:Complement Factor H; D010958:Plasminogen; D005341:Fibrinolysin