1887

Abstract

is an important cause of different pathologies in pigs and humans, most importantly fibrinosuppurative meningitis. Tissue infected with this pathogen is substantially infiltrated with neutrophils, but the function of neutrophil extracellular traps (NETs) - a more recently discovered antimicrobial strategy of neutrophils - in host defence against has not been investigated. The objective of this work was to investigate the interaction of with NETs . induced NET formation in porcine neutrophils and was entrapped but not killed by those NETs. As the amount of NETs decreased over time, we hypothesized that a known extracellular DNase of degrades NETs. Though this nuclease was originally designated -secreted nuclease A (SsnA), this work demonstrated surface association in accordance with an LPXTG cell wall anchor motif and partial release into the supernatant. Confirming our hypothesis, an isogenic A mutant was significantly attenuated in NET degradation and in protection against the antimicrobial activity of NETs as determined in assays with phorbol myristate acetate (PMA)-stimulated human neutrophils. Though assays with PMA-stimulated porcine neutrophils suggested that SsnA also degrades porcine NETs, phenotypic differences between wt and the isogenic A mutant were less distinct. As SsnA expression was crucial for neither growth nor for survival in porcine or human blood, the results indicated that SsnA is the first specific NET evasion factor to be identified in .

Funding
This study was supported by the:
  • Ministry of Science and Culture of Lower Saxony
  • Akademie für Tiergesundheit, Germany
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2014-02-01
2024-03-29
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References

  1. Arends J. P., Zanen H. C. ( 1988). Meningitis caused by Streptococcus suis in humans. Rev Infect Dis 10:131–137 [View Article][PubMed]
    [Google Scholar]
  2. Baums C. G., Valentin-Weigand P. ( 2009). Surface-associated and secreted factors of Streptococcus suis in epidemiology, pathogenesis and vaccine development. Anim Health Res Rev 10:65–83 [View Article][PubMed]
    [Google Scholar]
  3. Baums C. G., Kaim U., Fulde M., Ramachandran G., Goethe R., Valentin-Weigand P. ( 2006). Identification of a novel virulence determinant with serum opacification activity in Streptococcus suis.. Infect Immun 74:6154–6162 [View Article][PubMed]
    [Google Scholar]
  4. Baums C. G., Kock C., Beineke A., Bennecke K., Goethe R., Schröder C., Waldmann K. H., Valentin-Weigand P. ( 2009). Streptococcus suis bacterin and subunit vaccine immunogenicities and protective efficacies against serotypes 2 and 9. Clin Vaccine Immunol 16:200–208 [View Article][PubMed]
    [Google Scholar]
  5. Beineke A., Bennecke K., Neis C., Schröder C., Waldmann K. H., Baumgärtner W., Valentin-Weigand P., Baums C. G. ( 2008). Comparative evaluation of virulence and pathology of Streptococcus suis serotypes 2 and 9 in experimentally infected growers. Vet Microbiol 128:423–430 [View Article][PubMed]
    [Google Scholar]
  6. Beiter K., Wartha F., Albiger B., Normark S., Zychlinsky A., Henriques-Normark B. ( 2006). An endonuclease allows Streptococcus pneumoniae to escape from neutrophil extracellular traps. Curr Biol 16:401–407 [View Article][PubMed]
    [Google Scholar]
  7. Benga L., Fulde M., Neis C., Goethe R., Valentin-Weigand P. ( 2008). Polysaccharide capsule and suilysin contribute to extracellular survival of Streptococcus suis co-cultivated with primary porcine phagocytes. Vet Microbiol 132:211–219 [View Article][PubMed]
    [Google Scholar]
  8. Berends E. T., Horswill A. R., Haste N. M., Monestier M., Nizet V., von Köckritz-Blickwede M. ( 2010). Nuclease expression by Staphylococcus aureus facilitates escape from neutrophil extracellular traps. J Innate Immun 2:576–586 [View Article][PubMed]
    [Google Scholar]
  9. Bréa D., Meurens F., Dubois A. V., Gaillard J., Chevaleyre C., Jourdan M.-L., Winter N., Arbeille B., Si-Tahar M. & other authors ( 2012). The pig as a model for investigating the role of neutrophil serine proteases in human inflammatory lung diseases. Biochem J 447:363–370 [View Article][PubMed]
    [Google Scholar]
  10. Brinkmann V., Reichard U., Goosmann C., Fauler B., Uhlemann Y., Weiss D. S., Weinrauch Y., Zychlinsky A. ( 2004). Neutrophil extracellular traps kill bacteria. Science 303:1532–1535 [View Article][PubMed]
    [Google Scholar]
  11. Buchanan J. T., Simpson A. J., Aziz R. K., Liu G. Y., Kristian S. A., Kotb M., Feramisco J., Nizet V. ( 2006). DNase expression allows the pathogen group A Streptococcus to escape killing in neutrophil extracellular traps. Curr Biol 16:396–400 [View Article][PubMed]
    [Google Scholar]
  12. Chabot-Roy G., Willson P., Segura M., Lacouture S., Gottschalk M. ( 2006). Phagocytosis and killing of Streptococcus suis by porcine neutrophils. Microb Pathog 41:21–32 [View Article][PubMed]
    [Google Scholar]
  13. Chang A., Khemlani A., Kang H., Proft T. ( 2011). Functional analysis of Streptococcus pyogenes nuclease A (SpnA), a novel group A streptococcal virulence factor. Mol Microbiol 79:1629–1642 [View Article][PubMed]
    [Google Scholar]
  14. Charland N., Harel J., Kobisch M., Lacasse S., Gottschalk M. ( 1998). Streptococcus suis serotype 2 mutants deficient in capsular expression. Microbiology 144:325–332 [View Article][PubMed]
    [Google Scholar]
  15. Chow O. A., von Köckritz-Blickwede M., Bright A. T., Hensler M. E., Zinkernagel A. S., Cogen A. L., Gallo R. L., Monestier M., Wang Y. & other authors ( 2010). Statins enhance formation of phagocyte extracellular traps. Cell Host Microbe 8:445–454 [View Article][PubMed]
    [Google Scholar]
  16. Derré-Bobillot A., Cortes-Perez N. G., Yamamoto Y., Kharrat P., Couvé E., Da Cunha V., Decker P., Boissier M.-C., Escartin F. & other authors ( 2013). Nuclease A (Gbs0661), an extracellular nuclease of Streptococcus agalactiae, attacks the neutrophil extracellular traps and is needed for full virulence. Mol Microbiol 89:518–531 [View Article][PubMed]
    [Google Scholar]
  17. Fittipaldi N., Sekizaki T., Takamatsu D., Harel J., Domínguez-Punaro M. L., Von Aulock S., Draing C., Marois C., Kobisch M., Gottschalk M. ( 2008). d-alanylation of lipoteichoic acid contributes to the virulence of Streptococcus suis.. Infect Immun 76:3587–3594 [View Article][PubMed]
    [Google Scholar]
  18. Fontaine M. C., Perez-Casal J., Willson P. J. ( 2004). Investigation of a novel DNase of Streptococcus suis serotype 2. Infect Immun 72:774–781 [View Article][PubMed]
    [Google Scholar]
  19. Fuchs T. A., Abed U., Goosmann C., Hurwitz R., Schulze I., Wahn V., Weinrauch Y., Brinkmann V., Zychlinsky A. ( 2007). Novel cell death program leads to neutrophil extracellular traps. J Cell Biol 176:231–241 [View Article][PubMed]
    [Google Scholar]
  20. Gómez-Gascón L., Luque I., Olaya-Abril A., Jiménez-Munguía I., Orbegozo-Medina R. A., Peralbo E., Tarradas C., Rodríguez-Ortega M. J. ( 2012). Exploring the pan-surfome of Streptococcus suis: looking for common protein antigens. J Proteomics 75:5654–5666 [View Article][PubMed]
    [Google Scholar]
  21. Gottschalk M. ( 2011). Streptococcosis. Diseases of Swine841–855 Zimmerman J. J., Kariker L. A., Ramirez A., Schwartz K. J., Stevenson G. W. Oxford, UK: Wiley-Blackwell;
    [Google Scholar]
  22. Gottschalk M., Xu J., Calzas C., Segura M. ( 2010). Streptococcus suis: a new emerging or an old neglected zoonotic pathogen. Future Microbiol 5:371–391 [View Article][PubMed]
    [Google Scholar]
  23. Lappann M., Danhof S., Guenther F., Olivares-Florez S., Mordhorst I. L., Vogel U. ( 2013). In vitro resistance mechanisms of Neisseria meningitidis against neutrophil extracellular traps. Mol Microbiol 89:433–449 [View Article][PubMed]
    [Google Scholar]
  24. Lauth X., von Köckritz-Blickwede M., McNamara C. W., Myskowski S., Zinkernagel A. S., Beall B., Ghosh P., Gallo R. L., Nizet V. ( 2009). M1 protein allows Group A streptococcal survival in phagocyte extracellular traps through cathelicidin inhibition. J Innate Immun 1:202–214 [View Article][PubMed]
    [Google Scholar]
  25. Losman M. J., Fasy T. M., Novick K. E., Monestier M. ( 1992). Monoclonal autoantibodies to subnucleosomes from a MRL/Mp(-)+/+ mouse. Oligoclonality of the antibody response and recognition of a determinant composed of histones H2A, H2B, and DNA. J Immunol 148:1561–1569[PubMed]
    [Google Scholar]
  26. Lun Z. R., Wang Q. P., Chen X. G., Li A. X., Zhu X. Q. ( 2007). Streptococcus suis: an emerging zoonotic pathogen. Lancet Infect Dis 7:201–209 [View Article][PubMed]
    [Google Scholar]
  27. Mai N. T., Hoa N. T., Nga T. V., Linh D., Chau T. T., Sinh D. X., Phu N. H., Chuong L. V., Diep T. S. & other authors ( 2008). Streptococcus suis meningitis in adults in Vietnam. Clin Infect Dis 46:659–667 [View Article][PubMed]
    [Google Scholar]
  28. Midon M., Schäfer P., Pingoud A., Ghosh M., Moon A. F., Cuneo M. J., London R. E., Meiss G. ( 2011). Mutational and biochemical analysis of the DNA-entry nuclease EndA from Streptococcus pneumoniae.. Nucleic Acids Res 39:623–634 [View Article][PubMed]
    [Google Scholar]
  29. Pilsczek F. H., Salina D., Poon K. K., Fahey C., Yipp B. G., Sibley C. D., Robbins S. M., Green F. H., Surette M. G. & other authors ( 2010). A novel mechanism of rapid nuclear neutrophil extracellular trap formation in response to Staphylococcus aureus.. J Immunol 185:7413–7425 [View Article][PubMed]
    [Google Scholar]
  30. Sambrook J., Fritsch E. F., Maniatis T. ( 1989). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  31. Scapinello S., Brooks A. S., MacInnes J. I., Hammermueller J., Clark M. E., Caswell J. L. ( 2011). Bactericidal activity of porcine neutrophil secretions. Vet Immunol Immunopathol 139:113–118 [View Article][PubMed]
    [Google Scholar]
  32. Seele J., Singpiel A., Spoerry C., von Pawel-Rammingen U., Valentin-Weigand P., Baums C. G. ( 2013). Identification of a novel host-specific IgM protease in Streptococcus suis.. J Bacteriol 195:930–940 [View Article][PubMed]
    [Google Scholar]
  33. Smith H. E., Damman M., van der Velde J., Wagenaar F., Wisselink H. J., Stockhofe-Zurwieden N., Smits M. A. ( 1999). Identification and characterization of the cps locus of Streptococcus suis serotype 2: the capsule protects against phagocytosis and is an important virulence factor. Infect Immun 67:1750–1756[PubMed]
    [Google Scholar]
  34. Takamatsu D., Osaki M., Sekizaki T. ( 2001). Thermosensitive suicide vectors for gene replacement in Streptococcus suis.. Plasmid 46:140–148 [View Article][PubMed]
    [Google Scholar]
  35. Tang J., Wang C., Feng Y., Yang W., Song H., Chen Z., Yu H., Pan X., Zhou X. & other authors ( 2006). Streptococcal toxic shock syndrome caused by Streptococcus suis serotype 2. PLoS Med 3:e151 [View Article][PubMed]
    [Google Scholar]
  36. Vasconcelos D., Middleton D. M., Chirino-Trejo J. M. ( 1994). Lesions caused by natural infection with Streptococcus suis type 9 in weaned pigs. J Vet Diagn Invest 6:335–341 [View Article][PubMed]
    [Google Scholar]
  37. von Köckritz-Blickwede M., Nizet V. ( 2009). Innate immunity turned inside-out: antimicrobial defense by phagocyte extracellular traps. J Mol Med (Berl) 87:775–783 [View Article][PubMed]
    [Google Scholar]
  38. von Köckritz-Blickwede M., Chow O., Ghochani M., Nizet V. ( 2010). Visualization and functional evaluation of phagocyte extracellular traps. Methods in Microbiology138–160 Kaufmann S. H., Kabelitz D. London, UK: Academic Press; [View Article]
    [Google Scholar]
  39. Wangkaew S., Chaiwarith R., Tharavichitkul P., Supparatpinyo K. ( 2006). Streptococcus suis infection: a series of 41 cases from Chiang Mai University Hospital. J Infect 52:455–460 [View Article][PubMed]
    [Google Scholar]
  40. Wartha F., Beiter K., Albiger B., Fernebro J., Zychlinsky A., Normark S., Henriques-Normark B. ( 2007). Capsule and d-alanylated lipoteichoic acids protect Streptococcus pneumoniae against neutrophil extracellular traps. Cell Microbiol 9:1162–1171 [View Article][PubMed]
    [Google Scholar]
  41. Wei Z., Li R., Zhang A., He H., Hua Y., Xia J., Cai X., Chen H., Jin M. ( 2009). Characterization of Streptococcus suis isolates from the diseased pigs in China between 2003 and 2007. Vet Microbiol 137:196–201 [View Article][PubMed]
    [Google Scholar]
  42. Willenborg J., Fulde M., de Greeff A., Rohde M., Smith H. E., Valentin-Weigand P., Goethe R. ( 2011). Role of glucose and CcpA in capsule expression and virulence of Streptococcus suis.. Microbiology 157:572–582 [View Article][PubMed]
    [Google Scholar]
  43. Williams A. E., Blakemore W. F. ( 1990). Pathogenesis of meningitis caused by Streptococcus suis type 2. J Infect Dis 162:474–481 [View Article][PubMed]
    [Google Scholar]
  44. Wisselink H. J., Smith H. E., Stockhofe-Zurwieden N., Peperkamp K., Vecht U. ( 2000). Distribution of capsular types and production of muramidase-released protein (MRP) and extracellular factor (EF) of Streptococcus suis strains isolated from diseased pigs in seven European countries. Vet Microbiol 74:237–248 [View Article][PubMed]
    [Google Scholar]
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