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Evaluation of the Expression and Protective Potential of Leptospiral Sphingomyelinases

Current Microbiology volume 60, pages 134–142 (2010)Cite this article

Abstract

Leptospirosis is a zoonotic disease of global distribution, which affects both animals and humans. Pathogenic leptospires, the bacteria that cause this disease, require iron for their growth, and these spirochetes probably use their hemolysins, such as the sphingomyelinases, as a way to obtain this important nutrient from host red blood cells during infection. We expressed and purified the leptospiral sphingomyelinases Sph1, Sph2, Sph4, and SphH in a heterologous system. However, the recombinant proteins were not able to lyse sheep erythrocytes, despite having regular secondary structures. Transcripts for all sphingomyelinases tested were detected by RT-PCR analyses, but only Sph2 and SphH native proteins could be detected in Western blot assays using Leptospira whole extracts as well as in renal tubules of infected hamsters. Moreover, antibodies present in the serum of a human patient with laboratory-confirmed leptospirosis recognized Sph2, indicating that this sphingomyelinase is expressed and exposed to the immune system during infection in humans. However, in an animal challenge model, none of the sphingomyelinases tested conferred protection against leptospirosis.

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Acknowledgments

We thank Dr. Albert Leyva for English editing of the manuscript and FAPESP, CNPq, and Fundação Butantan for financial support.

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Affiliations

  1. Centro de Biotecnologia, Instituto Butantan, 05503-900, São Paulo, SP, Brazil

    Eneas Carvalho, Maria L. S. Oliveira & Paulo L. Ho

  2. Departamento de Genética e Biologia Evolutiva, Instituto de Biociências, Universidade de São Paulo, São Paulo, SP, Brazil

    Eneas Carvalho & Paulo L. Ho

  3. Laboratório de Parasitologia, Instituto Butantan, São Paulo, SP, Brazil

    Eneas Carvalho

  4. Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brazil

    Angela S. Barbosa

  5. Instituto de Biotecnología y Biología Molecular, Universidad Nacional de La Plata, Centro Científico Tecnológico CONICET, La Plata, Argentina

    Ricardo M. Gómez

  6. Seção de Bacteriologia, Instituto Adolfo Lutz, São Paulo, SP, Brazil

    Eliete C. Romero

  7. Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP, Brazil

    Amane P. Gonçales, Zenaide M. Morais & Sílvio A. Vasconcellos

Authors
  1. Eneas Carvalho
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  2. Angela S. Barbosa
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  3. Ricardo M. Gómez
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  4. Maria L. S. Oliveira
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  5. Eliete C. Romero
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  6. Amane P. Gonçales
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  7. Zenaide M. Morais
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  8. Sílvio A. Vasconcellos
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  9. Paulo L. Ho
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Correspondence to Paulo L. Ho.

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Carvalho, E., Barbosa, A.S., Gómez, R.M. et al. Evaluation of the Expression and Protective Potential of Leptospiral Sphingomyelinases. Curr Microbiol 60, 134–142 (2010). https://doi.org/10.1007/s00284-009-9519-3

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  • DOI: https://doi.org/10.1007/s00284-009-9519-3

Keywords

  • Recombinant Protein
  • Hemolytic Activity
  • Leptospirosis
  • Regular Secondary Structure
  • Pathogenic Leptospire