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Leptospira interrogans Bat proteins impair host hemostasis by fibrinogen cleavage and platelet aggregation inhibition

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Abstract

Leptospirosis is a worldwide spread zoonosis, caused by pathogenic Leptospira. Evidences suggest that compromised hemostasis might be involved in the leptospirosis pathophysiology. In the genome of L. interrogans serovar Copenhageni, we found two genes coding for proteins which comprise von Willebrand factor (VWF) A domains (BatA and BatB). As VWF A domains exhibit multiple binding sites which contributes to human VWF hemostatic functions, we hypothesized that the L. interrogans BatA and BatB proteins could be involved in the hemostatic impairment during leptospirosis. We have cloned, expressed in Escherichia coli, and purified recombinant BatA and BatB. The influence of recombinant BatA and BatB on different in vitro hemostatic assays evaluating the enzymatic activity, platelet aggregation and fibrinogen integrity was investigated. We describe BatB as a new serine protease which is able to cleave thrombin chromogenic substrate, fibrin, fibrinogen, gelatin and casein; while BatA is active only towards fibrinogen. BatA and BatB interfere with the platelet aggregation induced by VWF/ristocetin and thrombin. Our results suggest an important role of the L. interrogans serovar Copenhageni Bat proteins in the hemostasis dysfunction observed during leptospirosis and contribute to the understanding of the leptospirosis pathophysiological mechanisms.

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Acknowledgements

We thank Gisele de Souza for excellent technical assistance.

Funding

This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Brazil [2017/00236-5; 2014/50981-0; 2018/07054-2 to M.L.V.; 2017/01102-2 to F.J.P.] and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) to M.B.H. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Author notes

  1. Mônica Larucci Vieira

    Present address: Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil

  2. Ana Lucia T. O. Nascimento and Mônica Larucci Vieira share senior authorship.

Authors and Affiliations

  1. Lab. Desenvolvimento de Vacinas, Instituto Butantan, São Paulo, Brazil

    Felipe José Passalia, Ana Lucia T. O. Nascimento & Mônica Larucci Vieira

  2. Programa de Pós-Graduação Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil

    Felipe José Passalia & Ana Lucia T. O. Nascimento

  3. Lab. de Zoonoses Bacterianas, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, Brazil

    Marcos Bryan Heinemann

  4. Lab. Especial de Dor e Sinalização Celular, Instituto Butantan, São Paulo, Brazil

    Sonia Aparecida de Andrade

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  1. Felipe José Passalia
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Contributions

Conceived and designed the experiments: MV and FP. Performed the experiments: MV and FP. Analyzed the data: MV and FP. Contributed with reagents/materials/analysis tools: MV, MH, SA and AN. Wrote the paper: MV. Revised the paper: all authors.

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Correspondence to Mônica Larucci Vieira.

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Passalia, F.J., Heinemann, M.B., de Andrade, S.A. et al. Leptospira interrogans Bat proteins impair host hemostasis by fibrinogen cleavage and platelet aggregation inhibition. Med Microbiol Immunol 209, 201–213 (2020). https://doi.org/10.1007/s00430-020-00664-4

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