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Abstract

Leptospirosis is a widespread zoonosis caused by invasive spirochaetes belonging to the genus . Pathogenic leptospires disseminate via the bloodstream to colonize the renal tubules of reservoir hosts. Little is known about leptospiral outer-membrane proteins expressed during the dissemination stage of infection. In this study, a novel surface-exposed lipoprotein is described; it has been designated LipL46 to distinguish it from a previously described 31 kDa peripheral membrane protein, P31, which is exported as a 45 kDa probable lipoprotein. The gene encodes a 412 aa polypeptide with a 21 aa signal peptide. Lipid modification of cysteine at the lipoprotein signal peptidase cleavage site FSISC is supported by the finding that intrinsically labels LipL46 during incubation in medium containing [C]palmitate. LipL46 appears to be exported to the leptospiral outer membrane as a 46 kDa lipoprotein, based on Triton X-114 solubilization and phase partitioning studies, which included the outer and inner membrane controls LipL32 and LipL31, respectively. Surface immunoprecipitation and whole-cell ELISA experiments indicate that LipL46 is exposed on the leptospiral surface. Immunohistochemistry studies demonstrated expression of LipL46 by leptospires found in the bloodstream of acutely infected hamsters. Leptospires expressing LipL46 were also found in the intercellular spaces of the liver, within splenic phagocytes, and invading the glomerular hilum of the kidney. Infection-associated expression is supported by the finding that LipL46 is a major antigen recognized by sera from infected hamsters. These findings indicate that LipL46 may be important in leptospiral dissemination, and that it may serve as a useful serodiagnostic antigen.

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2006-12-01
2019-10-15
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References

  1. Babudieri, B. ( 1958; ). Animal reservoirs of leptospires. Ann N Y Acad Sci 70, 393–413.[CrossRef]
    [Google Scholar]
  2. Barnett, J. K., Barnett, D., Bolin, C. A., Summers, T. A., Wagar, E. A., Cheville, N. F., Hartskeerl, R. A. & Haake, D. A. ( 1999; ). Expression and distribution of leptospiral outer membrane components during renal infection of hamsters. Infect Immun 67, 853–861.
    [Google Scholar]
  3. Branger, C., Sonrier, C., Chatrenet, B., Klonjkowski, B., Ruvoen-Clouet, N., Aubert, A., Andre-Fontaine, G. & Eloit, M. ( 2001; ). Identification of the hemolysis-associated protein 1 as a cross-protective immunogen of Leptospira interrogans by adenovirus-mediated vaccination. Infect Immun 69, 6831–6838.[CrossRef]
    [Google Scholar]
  4. Cullen, P. A., Cordwell, S. J., Bulach, D. M., Haake, D. A. & Adler, B. ( 2002; ). Global analysis of outer membrane proteins from Leptospira interrogans serovar Lai. Infect Immun 70, 2311–2318.[CrossRef]
    [Google Scholar]
  5. Cullen, P. A., Haake, D. A., Bulach, D. M., Zuerner, R. L. & Adler, B. ( 2003; ). LipL21 is a novel surface-exposed lipoprotein of pathogenic Leptospira species. Infect Immun 71, 2414–2421.[CrossRef]
    [Google Scholar]
  6. Cullen, P. A., Xu, X., Matsunaga, J., Sanchez, Y., Ko, A. I., Haake, D. A. & Adler, B. ( 2005; ). Surfaceome of Leptospira spp. Infect Immun 73, 4853–4863.[CrossRef]
    [Google Scholar]
  7. Faine, S., Adler, B., Bolin, C. & Perolat, P. ( 1999; ). Leptospira and Leptospirosis, 2nd edn. Melbourne, Australia: MediSci.
  8. Guerreiro, H., Croda, J., Flannery, B., Mazel, M., Matsunaga, J., Galvao Reis, M., Levett, P. N., Ko, A. I. & Haake, D. A. ( 2001; ). Leptospiral proteins recognized during the humoral immune response to leptospirosis in humans. Infect Immun 69, 4958–4968.[CrossRef]
    [Google Scholar]
  9. Haake, D. A. ( 2000; ). Spirochaetal lipoproteins and pathogenesis. Microbiology 146, 1491–1504.
    [Google Scholar]
  10. Haake, D. A. & Matsunaga, J. ( 2002; ). Characterization of the leptospiral outer membrane and description of three novel leptospiral membrane proteins. Infect Immun 70, 4936–4945.[CrossRef]
    [Google Scholar]
  11. Haake, D. A., Walker, E. M., Blanco, D. R., Bolin, C. A., Miller, M. N. & Lovett, M. A. ( 1991; ). Changes in the surface of Leptospira interrogans serovar Grippotyphosa during in vitro cultivation. Infect Immun 59, 1131–1140.
    [Google Scholar]
  12. Haake, D. A., Champion, C. I., Martinich, C., Shang, E. S., Blanco, D. R., Miller, J. N. & Lovett, M. A. ( 1993; ). Molecular cloning and sequence analysis of the gene encoding OmpL1, a transmembrane outer membrane protein of pathogenic Leptospira spp. J Bacteriol 175, 4225–4234.
    [Google Scholar]
  13. Haake, D. A., Martinich, C., Summers, T. A., Shang, E. S., Pruetz, J. D., McCoy, A. M., Mazel, M. K. & Bolin, C. A. ( 1998; ). Characterization of leptospiral outer membrane lipoprotein LipL36: downregulation associated with late-log-phase growth and mammalian infection. Infect Immun 66, 1579–1587.
    [Google Scholar]
  14. Haake, D. A., Chao, G., Zuerner, R. L., Barnett, J. K., Barnett, D., Mazel, M., Matsunaga, J., Levett, P. N. & Bolin, C. A. ( 2000; ). The leptospiral major outer membrane protein LipL32 is a lipoprotein expressed during mammalian infection. Infect Immun 68, 2276–2285.[CrossRef]
    [Google Scholar]
  15. Haake, D. A., Suchard, M. A., Kelley, M. M., Dundoo, M., Alt, D. P. & Zuerner, R. L. ( 2004; ). Molecular evolution and mosaicism of leptospiral outer membrane proteins involves horizontal DNA transfer. J Bacteriol 186, 2818–2828.[CrossRef]
    [Google Scholar]
  16. Inada, R., Ido, Y., Hoki, R., Kaneko, R. & Ito, H. ( 1916; ). The etiology, mode of infection, and specific therapy of Weil's disease (spirochaetosis icterohaemorrhagica). J Exp Med 23, 377–402.[CrossRef]
    [Google Scholar]
  17. Johnson, R. C. & Harris, V. G. ( 1967; ). Differentiation of pathogenic and saprophytic letospires. I. Growth at low temperatures. J Bacteriol 94, 27–31.
    [Google Scholar]
  18. Juncker, A. S., Willenbrock, H., Von Heijne, G., Brunak, S., Nielsen, H. & Krogh, A. ( 2003; ). Prediction of lipoprotein signal peptides in Gram-negative bacteria. Protein Sci 12, 1652–1662.[CrossRef]
    [Google Scholar]
  19. Koizumi, N. & Watanabe, H. ( 2003; ). Molecular cloning and characterization of a novel leptospiral lipoprotein with OmpA domain. FEMS Microbiol Lett 226, 215–219.[CrossRef]
    [Google Scholar]
  20. Levett, P. N. ( 2001; ). Leptospirosis. Clin Microbiol Rev 14, 296–326.[CrossRef]
    [Google Scholar]
  21. Matsunaga, J., Young, T. A., Barnett, J. K., Barnett, D., Bolin, C. A. & Haake, D. A. ( 2002; ). Novel 45-kilodalton leptospiral protein that is processed to a 31-kilodalton growth-phase-regulated peripheral membrane protein. Infect Immun 70, 323–334.[CrossRef]
    [Google Scholar]
  22. Matsunaga, J., Barocchi, M. A., Croda, J. & 8 other authors ( 2003; ). Pathogenic Leptospira species express surface-exposed proteins belonging to the bacterial immunoglobulin superfamily. Mol Microbiol 49, 929–945.[CrossRef]
    [Google Scholar]
  23. Matsunaga, J., Sanchez, Y., Xu, X. & Haake, D. A. ( 2005; ). Osmolarity, a key environmental signal controlling expression of leptospiral proteins LigA and LigB and the extracellular release of LigA. Infect Immun 73, 70–78.[CrossRef]
    [Google Scholar]
  24. McBride, A. J., Athanazio, D. A., Reis, M. G. & Ko, A. I. ( 2005; ). Leptospirosis. Curr Opin Infect Dis 18, 376–386.[CrossRef]
    [Google Scholar]
  25. Merien, F., Baranton, G. & Perolat, P. ( 1997; ). Invasion of Vero cells and induction of apoptosis in macrophages by pathogenic Leptospira interrogans are correlated with virulence. Infect Immun 65, 729–738.
    [Google Scholar]
  26. Nally, J. E., Chow, E., Fishbein, M. C., Blanco, D. R. & Lovett, M. A. ( 2005a; ). Changes in lipopolysaccharide O antigen distinguish acute versus chronic Leptospira interrogans infections. Infect Immun 73, 3251–3260.[CrossRef]
    [Google Scholar]
  27. Nally, J. E., Whitelegge, J. P., Aguilera, R., Pereira, M. M., Blanco, D. R. & Lovett, M. A. ( 2005b; ). Purification and proteomic analysis of outer membrane vesicles from a clinical isolate of Leptospira interrogans serovar Copenhageni. Proteomics 5, 144–152.[CrossRef]
    [Google Scholar]
  28. Nascimento, A. L., Ko, A. I., Martins, E. A. & 44 other authors ( 2004; ). Comparative genomics of two Leptospira interrogans serovars reveals novel insights into physiology and pathogenesis. J Bacteriol 186, 2164–2172.[CrossRef]
    [Google Scholar]
  29. Norris, S. J., Carter, C. J., Howell, J. K. & Barbour, A. G. ( 1992; ). Low-passage-associated proteins of Borrelia burgdorferi B31: characterization and molecular cloning of OspD, a surface-exposed, plasmid-encoded lipoprotein. Infect Immun 60, 4662–4672.
    [Google Scholar]
  30. Ren, S. X., Fu, G., Jiang, X. G. & 36 other authors ( 2003; ). Unique physiological and pathogenic features of Leptospira interrogans revealed by whole-genome sequencing. Nature 422, 888–893.[CrossRef]
    [Google Scholar]
  31. Schulze, R. J. & Zuckert, W. R. ( 2006; ). Borrelia burgdorferi lipoproteins are secreted to the outer surface by default. Mol Microbiol 59, 1473–1484.[CrossRef]
    [Google Scholar]
  32. Segura, E. R., Ganoza, C. A., Campos, K. & 11 other authors ( 2005; ). Clinical spectrum of pulmonary involvement in leptospirosis in a region of endemicity, with quantification of leptospiral burden. Clin Infect Dis 40, 343–351.[CrossRef]
    [Google Scholar]
  33. Setubal, J. C., Reis, M. G., Matsunaga, J. & Haake, D. ( 2006; ). Lipoprotein computational prediction in spirochaetal genomes. Microbiology 152, 113–121.[CrossRef]
    [Google Scholar]
  34. Shang, E. S., Exner, M. M., Summers, T. A., Martinich, C., Champion, C. I., Hancock, R. E. & Haake, D. A. ( 1995; ). The rare outer membrane protein, OmpL1, of pathogenic Leptospira species is a heat-modifiable porin. Infect Immun 63, 3174–3181.
    [Google Scholar]
  35. Shang, E. S., Summers, T. A. & Haake, D. A. ( 1996; ). Molecular cloning and sequence analysis of the gene encoding LipL41, a surface-exposed lipoprotein of pathogenic Leptospira species. Infect Immun 64, 2322–2330.
    [Google Scholar]
  36. Steiner, G. & Steiner, G. ( 1944; ). New simple silver stain for demonstration of bacteria, spirochetes and fungi in sections of paraffin embedded tissue blocks. J Lab Clin Med 29, 868–871.
    [Google Scholar]
  37. Thiermann, A. B., McClellan, R. D. & Hill, H. T. ( 1984; ). Improved techniques for the isolation of leptospires from swine abortion cases. Ann Proc Am Assoc Vet Lab Diagn 27, 233–244.
    [Google Scholar]
  38. Verma, A., Artiushin, S., Matsunaga, J., Haake, D. A. & Timoney, J. F. ( 2005; ). LruA and LruB, novel lipoproteins of pathogenic Leptospira interrogans associated with equine recurrent uveitis. Infect Immun 73, 7259–7266.[CrossRef]
    [Google Scholar]
  39. Zuerner, R. L., Knudtson, W., Bolin, C. A. & Trueba, G. ( 1991; ). Characterization of outer membrane and secreted proteins of Leptospira interrogans serovar Pomona. Microb Pathog 10, 311–322.[CrossRef]
    [Google Scholar]
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vol. , part 12, pp. 3777-3786

Figs S1-S5 showing serial sections of hamster liver with and without leptospiral infection are available hereas an Acrobat PDF file.



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