Kochi Leandro Toshio, Fernandes Luis Guilherme Virgílio, Nascimento Ana Lucia Tabet Oller
Laboratório de Desenvolvimento de Vacinas, Instituto Butantan, São Paulo 05503-900, Brazil.
Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo 05508-900, Brazil.
Pathogens. 2020 Jul 22;9(8):599. doi: 10.3390/pathogens9080599.
Leptospirosis is a febrile disease and the etiological agents are pathogenic bacteria of the genus . The leptospiral virulence mechanisms are not fully understood and the application of genetic tools is still limited, despite advances in molecular biology techniques. The leptospiral recombinant protein LIC11711 has shown interaction with several host components, indicating a potential function in virulence. This study describes a system for heterologous expression of the gene using the saprophyte serovar Patoc as a surrogate, aiming to investigate its possible activity in bacterial virulence. Heterologous expression of LIC11711 was performed using the pMaOri vector under regulation of the promoter. The protein was found mainly on the leptospiral outer surface, confirming its location. The promoter enhanced the expression of LIC11711 in compared to the pathogenic strain, indicating that this strategy may be used to overexpress low-copy proteins. The presence of LIC11711 enhanced the capacity of to adhere to laminin (Lam) and plasminogen (Plg)/plasmin (Pla) in vitro, suggesting the involvement of this protein in bacterial pathogenesis. We show for the first time that the expression of LIC11711 protein of confers a virulence-associated phenotype on , pointing out possible mechanisms used by pathogenic leptospires.
钩端螺旋体病是一种发热性疾病，病原体是钩端螺旋体属的致病细菌。尽管分子生物学技术取得了进展，但钩端螺旋体的毒力机制尚未完全了解，遗传工具的应用仍然有限。钩端螺旋体重组蛋白LIC11711已显示与几种宿主成分相互作用，表明其在毒力方面具有潜在功能。本研究描述了一种利用腐生型帕托斯血清型作为替代物进行该基因异源表达的系统，旨在研究其在细菌毒力中的可能活性。使用pMaOri载体在启动子调控下进行LIC11711的异源表达。发现该蛋白主要位于钩端螺旋体外表面，证实了其定位。与致病菌株相比，启动子增强了LIC11711在中的表达，表明该策略可用于过表达低拷贝蛋白。LIC11711的存在增强了在体外黏附层粘连蛋白（Lam）和纤溶酶原（Plg）/纤溶酶（Pla）的能力，提示该蛋白参与细菌致病过程。我们首次表明，钩端螺旋体LIC11711蛋白的表达赋予了一种与毒力相关的表型，指出了致病钩端螺旋体可能使用的机制。
