Heterologous Expression of the Pathogen-Specific LIC11711 Gene in the Saprophyte L. biflexa Increases Bacterial Binding to Laminin and Plasminogen

Author(s)

Kochi, Leandro Toshio ^{[1, 2]} ; Fernandes, Luis Guilherme Virgilio ^[2] ; Nascimento, Ana Lucia Tabet Oller ^{[1, 2]}

Affiliation

^[1] Univ Sao Paulo, Inst Ciencias Biomed, BR-05508900 Sao Paulo - Brazil

^[2] Inst Butantan, Lab Desenvolvimento Vacinas, BR-05503900 Sao Paulo - Brazil

• Document type: Journal article
• Source: PATHOGENS; v. 9, n. 8 AUG 2020.
• Web of Science Citations: 0

Abstract

Leptospirosis is a febrile disease and the etiological agents are pathogenic bacteria of the genus Leptospira. 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 L. interrogans gene lic11711 using the saprophyte L. biflexa 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 lipL32 promoter. The protein was found mainly on the leptospiral outer surface, confirming its location. The lipL32 promoter enhanced the expression of LIC11711 in L. biflexa 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 L. biflexa 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 L. interrogans confers a virulence-associated phenotype on L. biflexa, pointing out possible mechanisms used by pathogenic leptospires. (AU)

• FAPESP's process: 18/09652-4 - Evaluation of the interaction of Leptospira interrogans proteins with host components by constructing recombinant Leptospira biflexa
• Grantee: Leandro Toshio Kochi
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 17/06731-8 - Application of CRISPR-interference for elucidating leptospirosis pathogenesis and development of novel strategies for knockout mutant's obtainment
• Grantee: Luis Guilherme Virgílio Fernandes
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/50981-0 - Search for surface proteins among the genome sequences of Leptospira interrogans: functional and immunological characterization to understanding mechanisms involved in the bacterial pathogenesis
• Grantee: Ana Lucia Tabet Oller do Nascimento
• Support Opportunities: Research Projects - Thematic Grants