Activation of human polymorphonuclear neutrophils by streptolysin O from Streptococcus pyogenes leads to the release of proinflammatory mediators

Summary

Streptococcus pyogenes is an important Gram-positive pathogen that is strictly limited to infections in humans. Here we report that streptolysin O (SLO),a cytolytic exotoxin secreted by S. pyogenes, activates human polymorphonuclear neutrophils (PMNs) by perforating these cells. This appears to be followed by an influx of Ca²⁺ and p38 MAPK activation. As a consequence, PMNs secrete heparin-binding protein, a potent inducer of vascular leakage, and neutrophil-borne proteins, including LL-37, α-defensins, and elastase. The results of the present work therefore suggest that the interaction between SLO and PMNs evokes an exaggerated host response which may contribute to the pathogenesis of local and generalized S. pyogenes infections.

Keywords

Infectious diseases - polymorphonuclear neutrophils - Streptococcus pyogenes - streptolysin O - degranulation

• References

• 1 Henderson B, Wilson M, Wren B. Are bacterial exotoxins cytokine network regulators?. Trends Microbiol 1997; 05: 454-8.
• 2 Bisno AL, Stevens DL. Streptococcal infections of skin and soft tissues. N Engl J Med 1996; 334: 240-5.
• 3 Herwald H, Cramer H, Mörgelin M. et al. M protein, a classical bacterial virulence determinant, forms complexes with fibrinogen that induce vascular leakage. Cell 2004; 116: 367-79.
• 4 Shafer WM, Martin LE, Spitznagel JK. Cationic antimicrobial proteins isolated from human neutrophil granulocytes in the presence of diisopropyl fluorophosphate. Infect Immun 1984; 45: 29-35.
• 5 Tapper H, Karlsson A, Mörgelin M. et al. Secretion of heparin-binding protein from human neutrophils is determined by its localization in azurophilic granules and secretory vesicles. Blood 2002; 99: 1785-93.
• 6 Pereira HA, Shafer WM, Pohl J. et al. CAP37, a human neutrophil-derived chemotactic factor with monocyte specific activity. J Clin Invest 1990; 85: 1468-76.
• 7 Gautam N, Olofsson AM, Herwald H. et al. Heparin-binding protein (HBP/CAP37): a missing link in neutrophil-evoked alteration of vascular permeability. Nat Med 2001; 07: 1123-7.
• 8 Ohno-Iwashita Y, Iwamoto M, Mitsui K. et al. Protease-nicked theta-toxin of Clostridium perfringens, a new membrane probe with no cytolytic effect, reveals two classes of cholesterol as toxin-binding sites on sheep erythrocytes. Eur J Biochem 1988; 176: 95-101.
• 9 Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 1977; 74: 5463-7.
• 10 Podbielski A, Spellerberg B, Woischnik M. et al. Novel series of plasmid vectors for gene inactivation and expression analysis in group A streptococci (GAS). Gene 1996; 177: 137-47.
• 11 Perez-Casal J, Price JA, Maguin E. et al. An M protein with a single C repeat prevents phagocytosis of Streptococcus pyogenes: use of a temperature-sensitive shuttle vector to deliver homologous sequences to the chromosome of S. pyogenes . Mol Microbiol 1993; 08: 809-19.
• 12 Perez-Casal J, Caparon MG, Scott JR. Mry, a transacting positive regulator of the M protein gene of Streptococcus pyogenes with similarity to the receptor proteins of two-component regulatory systems. J Bacteriol 1991; 173: 2617-24.
• 13 Thern A, Wästfelt M, Lindahl G. Expression of two different antiphagocytic M proteins by Streptococcus pyogenes of the OF+ lineage. J Immunol 1998; 160: 860-9.
• 14 Bricker AL, Cywes C, Ashbaugh CD. et al. NAD+-glycohydrolase acts as an intracellular toxin to enhance the extracellular survival of group A streptococci. Mol Microbiol 2002; 44: 257-69.
• 15 Baschong W, Lucocq JM, Roth J. “Thiocyanate gold”: small (2–3 nm) colloidal gold for affinity cytochemical labeling in electron microscopy. Histochemistry 1985; 83: 409-11.
• 16 Berge A, Kihlberg BM, Sjöholm AG. et al. Streptococcal proteinH forms soluble complement-activating complexes with IgG, but inhibits complement activation by IgG-coated targets. J Biol Chem 1997; 272: 20774-81.
• 17 Bhakdi S, Bayley H, Valeva A. et al. Staphylococcal alpha-toxin, streptolysin-O, and Escherichia coli hemolysin: prototypes of pore-forming bacterial cytolysins. Arch Microbiol 1996; 165: 73-9.
• 18 Borregaard N, Cowland JB. Granules of the human neutrophilic polymorphonuclear leukocyte. Blood 1997; 89: 3503-21.
• 19 Alouf JE. Streptococcal toxins (streptolysin O, streptolysin S, erythrogenic toxin). Pharmacol Ther 1980; 11: 661-717.
• 20 Sierig G, Cywes C, Wessels MR. et al. Cytotoxic effects of streptolysin o and streptolysin s enhance the virulence of poorly encapsulated group a streptococci. Infect Immun 2003; 71: 446-55.
• 21 Leino L, Forbes L, Segal A. et al. Reconstitution of GTPgammaS-induced NADPH oxidase activity in streptolysin-O-permeabilized neutrophils by specific cytosol fractions. Biochem Biophys Res Commun 1999; 265: 29-37.
• 22 Rosales JL, Ernst JD. Calcium-dependent neutrophil secretion: characterization and regulation by annexins. J Immunol 1997; 159: 6195-202.
• 23 Cuenda A, Rouse J, Doza YN. et al. SB 203580 is a specific inhibitor of a MAP kinase homologue which is stimulated by cellular stresses and interleukin-1. FEBS Lett 1995; 364: 229-33.
• 24 Chen JJ, Sun Y, Nabel GJ. Regulation of the proinflammatory effects of Fas ligand (CD95L). Science 1998; 282: 1714-7.
• 25 Cardenas ME, Sanfridson A, Cutler NS. et al. Signal-transduction cascades as targets for therapeutic intervention by natural products. Trends Biotechnol 1998; 16: 427-33.
• 26 Arcaro A, Wymann MP. Wortmannin is a potent phosphatidylinositol 3-kinase inhibitor: the role of phosphatidylinositol 3,4,5-trisphosphate in neutrophil responses. Biochem J 1993; 296: 297-301.
• 27 O’Dell TJ, Kandel ER, Grant SG. Long-term potentiation in the hippocampus is blocked by tyrosine kinase inhibitors. Nature 1991; 353: 558-60.
• 28 Fuortes M, Jin WW, Nathan C. Adhesion-dependent protein tyrosine phosphorylation in neutrophils treated with tumor necrosis factor. J Cell Biol 1993; 120: 777-84.
• 29 Dudley DT, Pang L, Decker SJ. et al. A synthetic inhibitor of the mitogen-activated protein kinase cascade. Proc Natl Acad Sci USA 1995; 92: 7686-9.
• 30 McLeish KR, Knall C, Ward RA. et al. Activation of mitogen-activated protein kinase cascades during priming of human neutrophils by TNF-alpha and GMCSF. J Leukoc Biol 1998; 64: 537-45.
• 31 Howard TH, Oresajo CO. A method for quantifying F-actin in chemotactic peptide activated neutrophils: study of the effect of tBOC peptide. Cell Motil 1985; 05: 545-57.
• 32 Dalpiaz A, Ferretti ME, Pecoraro R. et al. Phe-D-Leu-Phe-D-Leu-Phe derivatives as formylpeptide receptor antagonists in human neutrophils: cellular and conformational aspects. Biochim Biophys Acta 1999; 1432: 27-39.
• 33 Smallridge RC, Kiang JG, Gist ID. et al. U-73122, an aminosteroid phospholipase C antagonist, noncompetitively inhibits thyrotropin-releasing hormone effects in GH3 rat pituitary cells. Endocrinology 1992; 131: 1883-8.
• 34 Hellberg C, Molony L, Zheng L. et al. Ca2+ signalling mechanisms of the beta2 integrin on neutrophils: involvement of phospholipase C gamma 2 and Ins(1,4,5)P3. Biochem J 1996; 317: 403-9.
• 35 Stringaris AK, Geisenhainer J, Bergmann F. et al. Neurotoxicity of pneumolysin, a major pneumococcal virulence factor, involves calcium influx and depends on activation of p38 mitogen-activated protein kinase. Neurobiol Dis 2002; 11: 355-68.
• 36 Tilley SJ, Orlova EV, Gilbert RJ. et al. Structural basis of pore formation by the bacterial toxin pneumolysin. Cell 2005; 121: 247-56.
• 37 Cockeran R, Theron AJ, Steel HC. et al. Proinflammatory interactions of pneumolysin with human neutrophils. J Infect Dis 2001; 183: 604-11.
• 38 Elzi DJ, Bjornsen AJ, MacKenzie T. et al. Ionomycin causes activation of p38 and p42/44 mitogen-activated protein kinases in human neutrophils. Am J Physiol Cell Physiol 2001; 281: C350-60.
• 39 Sengelov H, Kjeldsen L, Borregaard N. Control of exocytosis in early neutrophil activation. J Immunol 1993; 150: 1535-43.
• 40 Ruiz N, Wang B, Pentland A. et al. Streptolysin O and adherence synergistically modulate proinflammatory responses of keratinocytes to group A streptococci. Mol Microbiol 1998; 27: 337-46.
• 41 Stassen M, Muller C, Richter C. et al. The streptococcal exotoxin streptolysin O activates mast cells to produce tumor necrosis factor alpha by p38 mitogenactivated protein kinase -and protein kinase C-dependent pathways. Infect Immun 2003; 71: 6171-7.
• 42 Mitsui K, Takano K, Nakatani S. et al. Chemokine production by rat macrophages stimulated with streptolysinO from Streptococcus pyogenes . Microbiol Immunol 2002; 46: 37-45.
• 43 Kawabata K, Hagio T, Matsuoka S. The role of neutrophil elastase in acute lung injury. Eur J Pharmacol 2002; 451: 1-10.
• 44 Tjabringa GS, Aarbiou J, Ninaber DK. et al. The antimicrobial peptide LL-37 activates innate immunity at the airway epithelial surface by transactivation of the epidermal growth factor receptor. J Immunol 2003; 171: 6690-6.
• 45 Scott MG, Davidson DJ, Gold MR. et al. The human antimicrobial peptide LL-37 is a multifunctional modulator of innate immune responses. J Immunol 2002; 169: 3883-91.