In LipL32, the Major Leptospiral Lipoprotein, the C Terminus Is the Primary Immunogenic Domain and Mediates Interaction with Collagen IV and Plasma Fibronectin

Abstract

LipL32 is the major leptospiral outer membrane lipoprotein expressed during infection and is the immu-

Calcium Binding to Leptospira Outer Membrane Antigen LipL32 Is Not Necessary for Its Interaction with Plasma Fibronectin, Collagen Type IV, and Plasminogen

“…Wild-type LipL32 with or without an N-terminal His tag fusion and a recombinant C-terminal fragment corresponding to residues 185-272 plus an N-terminal His tag (LipL32 185-272_His tag ) were produced as described (12,14). Mutations were introduced into the lipL32 gene by a PCR-driven overlap extension protocol (17) using as template the previously described plasmid lipL32ϩpAE (14).…”

“…LipL32 binding to these macromolecules was performed according to a previously published protocol with some modifications (12). Briefly, enzyme-linked immunosorbent assay (ELISA) plate wells (Nunc-Immuno Plate MaxiSorp Surface) were coated with 1 g of plasma fibronectin, F30, collagen type IV, or human plasminogen in 100 l of PBS and incubated overnight at 4°C.…”

“…Studies performed with recombinant LipL32 revealed that this highly immunogenic protein can induce a robust inflammatory response in renal proximal tubule cells (9,10). Furthermore, LipL32 may contribute to host colonization, because it has been demonstrated that it interacts with extracellular matrix (ECM) 3 components, such as collagen type IV, plasma fibronectin, and laminin (11,12) and binds human plasminogen (13).…”

Avaliação e caracterização de candidatos vacinais voltados para o controle da leptospirose.

“…Apesar disso, a ligação de LipL32 com laminina e colágeno foi demonstrado (HAUK et al, 2008;HOKE et al, 2008). Outra proteína interessante presente na membrana das leptospiras consiste na família de proteínas que apresentam domínios do tipo imunoglobulinas (Ig), nomeadas como LigA, LigB e LigC.…”

“…Várias proteínas de leptospiras com propriedades de ligação específica a laminina e a outros componentes da ECM foram identificadas pelo nosso grupo (ATZINGEN et al, 2008;ATZINGEN et al, 2009;BARBOSA et al, 2006;DOMINGOS et al, 2015;DOMINGOS et al, 2012;FERNANDES et al, 2014;FERNANDES et al, 2012;LONGHI et al, 2009;MENDES et al, 2011;OLIVEIRA et al, 2011;OLIVEIRA et al, 2010; __________________________________________________________Resultados e Discussão 113 2013; SOUZA et al, 2012;VIEIRA et al, 2010b). Outros grupos também identificaram proteínas com capacidade de adesão (CARVALHO et al, 2009;CHOY et al, 2007;HAUK et al, 2008;HOKE et al, 2008;LIN et al, 2009;STEVENSON et al, 2007). De maneira geral, esses dados demonstram o potencial que as leptospiras patogênicas apresentam em aderir nas células do hospedeiro a fim de estabelecer uma infecção.…”