Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1994 Sep;62(9):3731–3738. doi: 10.1128/iai.62.9.3731-3738.1994

Similar cytokine induction profiles of a novel streptococcal exotoxin, MF, and pyrogenic exotoxins A and B.

A Norrby-Teglund 1, M Norgren 1, S E Holm 1, U Andersson 1, J Andersson 1
PMCID: PMC303024  PMID: 8063387

Abstract

The cytokine production induced by a newly discovered streptococcal exotoxin, MF, and the pyrogenic exotoxins SpeA and SpeB was determined by in vitro stimulation of peripheral blood mononuclear cells (PBMCs) obtained from healthy blood donors. The induction and kinetics of interleukin-1 alpha (IL-1 alpha), IL-1 beta, IL-1 receptor antagonist, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-10, gamma interferon, tumor necrosis factor alpha (TNF-alpha), TNF-beta, and granulocyte-macrophage colony-stimulating factor were studied at the single-cell level by use of cytokine-specific monoclonal antibodies and intracellular immunofluorescent juxtanuclear staining. The cytokine-producing cells, with the exception of IL-1-expressing cells, had a characteristic morphology generated by the accumulation of cytokines in the Golgi organelle. MF, SpeA, and SpeB induced a massive gamma interferon and TNF-beta response in 10 to 16% of the PBMCs after 48 to 96 h of cell stimulation. In contrast, IL-2 and TNF-alpha production was detected in only 1 to 3% of the PBMCs. The induction of a lymphocyte TH2 phenotype response, including production of IL-3, IL-4, IL-5, and IL-10, was weak. However, the monokines, IL-1 alpha, IL-1 beta, IL-1 receptor antagonist, and IL-8, were consistently found and gradually produced, peaking at 24 h in approximately 5 to 8% of the PBMCs. MF showed extensive cytokine- and proliferation-inducing capacities equal to those of SpeA and SpeB, which suggests that MF is also a superantigen. A marked interindividual variation could be noted both in the proliferative response and in the cytokine induction of lymphocytes isolated from different individuals, which may be one explanation for the varying clinical severity noticed during group A streptococcal infections.

Full text

PDF
3731

Images in this article

3732Image
on p.3732
3733Image
on p.3733

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abrams J. S., Roncarolo M. G., Yssel H., Andersson U., Gleich G. J., Silver J. E. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992 Jun;127:5–24. doi: 10.1111/j.1600-065x.1992.tb01406.x. [DOI] [PubMed] [Google Scholar]
  2. Andersson J., Björk L., Dinarello C. A., Towbin H., Andersson U. Lipopolysaccharide induces human interleukin-1 receptor antagonist and interleukin-1 production in the same cell. Eur J Immunol. 1992 Oct;22(10):2617–2623. doi: 10.1002/eji.1830221022. [DOI] [PubMed] [Google Scholar]
  3. Andersson J., Nagy S., Björk L., Abrams J., Holm S., Andersson U. Bacterial toxin-induced cytokine production studied at the single-cell level. Immunol Rev. 1992 Jun;127:69–96. doi: 10.1111/j.1600-065x.1992.tb01409.x. [DOI] [PubMed] [Google Scholar]
  4. Andersson U., Matsuda T. Human interleukin 6 and tumor necrosis factor alpha production studied at a single-cell level. Eur J Immunol. 1989 Jun;19(6):1157–1160. doi: 10.1002/eji.1830190629. [DOI] [PubMed] [Google Scholar]
  5. Andersson U., Sander B. Detection of individual interleukin 2-producing cells after anti-CD3 antibody activation. Immunol Lett. 1989 Jan 31;20(2):115–120. doi: 10.1016/0165-2478(89)90095-3. [DOI] [PubMed] [Google Scholar]
  6. Barsumian E. L., Cunningham C. M., Schlievert P. M., Watson D. W. Heterogeneity of group A streptococcal pyrogenic exotoxin type B. Infect Immun. 1978 May;20(2):512–518. doi: 10.1128/iai.20.2.512-518.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bazzoni F., Cassatella M. A., Rossi F., Ceska M., Dewald B., Baggiolini M. Phagocytosing neutrophils produce and release high amounts of the neutrophil-activating peptide 1/interleukin 8. J Exp Med. 1991 Mar 1;173(3):771–774. doi: 10.1084/jem.173.3.771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Beutler B., Cerami A. The biology of cachectin/TNF--a primary mediator of the host response. Annu Rev Immunol. 1989;7:625–655. doi: 10.1146/annurev.iy.07.040189.003205. [DOI] [PubMed] [Google Scholar]
  9. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  10. Cunningham C. M., Barsumian E. L., Watson D. W. Further purification of group A streptococcal pyrogenic exotoxin and characterization of the purified toxin. Infect Immun. 1976 Sep;14(3):767–775. doi: 10.1128/iai.14.3.767-775.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cuturi M. C., Murphy M., Costa-Giomi M. P., Weinmann R., Perussia B., Trinchieri G. Independent regulation of tumor necrosis factor and lymphotoxin production by human peripheral blood lymphocytes. J Exp Med. 1987 Jun 1;165(6):1581–1594. doi: 10.1084/jem.165.6.1581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dolhain R. J., Andersson U., ter Haar N. T., Brinkman B. M., Verweij C. L., Daha M. R., Breedveld F. C., Miltenburg A. M. Detection of intracellular interferon-gamma by light microscopy using an immunoperoxidase technique: correlation with the corresponding mRNA and protein product. J Leukoc Biol. 1993 Dec;54(6):545–551. doi: 10.1002/jlb.54.6.545. [DOI] [PubMed] [Google Scholar]
  13. Fast D. J., Schlievert P. M., Nelson R. D. Toxic shock syndrome-associated staphylococcal and streptococcal pyrogenic toxins are potent inducers of tumor necrosis factor production. Infect Immun. 1989 Jan;57(1):291–294. doi: 10.1128/iai.57.1.291-294.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fleischer B., Gerardy-Schahn R., Metzroth B., Carrel S., Gerlach D., Köhler W. An evolutionary conserved mechanism of T cell activation by microbial toxins. Evidence for different affinities of T cell receptor-toxin interaction. J Immunol. 1991 Jan 1;146(1):11–17. [PubMed] [Google Scholar]
  15. Fleischer B., Schrezenmeier H., Conradt P. T lymphocyte activation by staphylococcal enterotoxins: role of class II molecules and T cell surface structures. Cell Immunol. 1989 Apr 15;120(1):92–101. doi: 10.1016/0008-8749(89)90177-9. [DOI] [PubMed] [Google Scholar]
  16. Fraser J. D. High-affinity binding of staphylococcal enterotoxins A and B to HLA-DR. Nature. 1989 May 18;339(6221):221–223. doi: 10.1038/339221a0. [DOI] [PubMed] [Google Scholar]
  17. Gardner S. M., Mock B. A., Hilgers J., Huppi K. E., Roeder W. D. Mouse lymphotoxin and tumor necrosis factor: structural analysis of the cloned genes, physical linkage, and chromosomal position. J Immunol. 1987 Jul 15;139(2):476–483. [PubMed] [Google Scholar]
  18. Gauchat J. F., Gauchat D., De Weck A. L., Stadler B. M. Cytokine mRNA levels in antigen-stimulated peripheral blood mononuclear cells. Eur J Immunol. 1989 Jun;19(6):1079–1085. doi: 10.1002/eji.1830190618. [DOI] [PubMed] [Google Scholar]
  19. Gerlach D., Knöll H., Köhler W. Isolierung und Charakterisierung von erythrogenen Toxinen. I. Untersuchung des von Streptococcus pyogenes, Stamm NY-5 gebildeten Toxins A. Zentralbl Bakteriol A. 1980;247(2):177–191. [PubMed] [Google Scholar]
  20. Gjörloff A., Fischer H., Hedlund G., Hansson J., Kenney J. S., Allison A. C., Sjögren H. O., Dohlsten M. Induction of interleukin-1 in human monocytes by the superantigen staphylococcal enterotoxin A requires the participation of T cells. Cell Immunol. 1991 Oct 1;137(1):61–71. doi: 10.1016/0008-8749(91)90056-h. [DOI] [PubMed] [Google Scholar]
  21. Goshorn S. C., Schlievert P. M. Nucleotide sequence of streptococcal pyrogenic exotoxin type C. Infect Immun. 1988 Sep;56(9):2518–2520. doi: 10.1128/iai.56.9.2518-2520.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Grossman D., Lamphear J. G., Mollick J. A., Betley M. J., Rich R. R. Dual roles for class II major histocompatibility complex molecules in staphylococcal enterotoxin-induced cytokine production and in vivo toxicity. Infect Immun. 1992 Dec;60(12):5190–5196. doi: 10.1128/iai.60.12.5190-5196.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hackett S. P., Stevens D. L. Streptococcal toxic shock syndrome: synthesis of tumor necrosis factor and interleukin-1 by monocytes stimulated with pyrogenic exotoxin A and streptolysin O. J Infect Dis. 1992 May;165(5):879–885. doi: 10.1093/infdis/165.5.879. [DOI] [PubMed] [Google Scholar]
  24. Hackett S. P., Stevens D. L. Superantigens associated with staphylococcal and streptococcal toxic shock syndrome are potent inducers of tumor necrosis factor-beta synthesis. J Infect Dis. 1993 Jul;168(1):232–235. doi: 10.1093/infdis/168.1.232. [DOI] [PubMed] [Google Scholar]
  25. Hauser A. R., Stevens D. L., Kaplan E. L., Schlievert P. M. Molecular analysis of pyrogenic exotoxins from Streptococcus pyogenes isolates associated with toxic shock-like syndrome. J Clin Microbiol. 1991 Aug;29(8):1562–1567. doi: 10.1128/jcm.29.8.1562-1567.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Herman A., Croteau G., Sekaly R. P., Kappler J., Marrack P. HLA-DR alleles differ in their ability to present staphylococcal enterotoxins to T cells. J Exp Med. 1990 Sep 1;172(3):709–717. doi: 10.1084/jem.172.3.709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Holm S. E., Norrby A., Bergholm A. M., Norgren M. Aspects of pathogenesis of serious group A streptococcal infections in Sweden, 1988-1989. J Infect Dis. 1992 Jul;166(1):31–37. doi: 10.1093/infdis/166.1.31. [DOI] [PubMed] [Google Scholar]
  28. Iwasaki M., Igarashi H., Hinuma Y., Yutsudo T. Cloning, characterization and overexpression of a Streptococcus pyogenes gene encoding a new type of mitogenic factor. FEBS Lett. 1993 Sep 27;331(1-2):187–192. doi: 10.1016/0014-5793(93)80323-m. [DOI] [PubMed] [Google Scholar]
  29. Janeway C. A., Jr, Yagi J., Conrad P. J., Katz M. E., Jones B., Vroegop S., Buxser S. T-cell responses to Mls and to bacterial proteins that mimic its behavior. Immunol Rev. 1989 Feb;107:61–88. doi: 10.1111/j.1600-065x.1989.tb00003.x. [DOI] [PubMed] [Google Scholar]
  30. Johnson L. P., Schlievert P. M., Watson D. W. Transfer of group A streptococcal pyrogenic exotoxin production to nontoxigenic strains of lysogenic conversion. Infect Immun. 1980 Apr;28(1):254–257. doi: 10.1128/iai.28.1.254-257.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Jupin C., Anderson S., Damais C., Alouf J. E., Parant M. Toxic shock syndrome toxin 1 as an inducer of human tumor necrosis factors and gamma interferon. J Exp Med. 1988 Mar 1;167(3):752–761. doi: 10.1084/jem.167.3.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kelso A. Frequency analysis of lymphokine-secreting CD4+ and CD8+ T cells activated in a graft-versus-host reaction. J Immunol. 1990 Oct 1;145(7):2167–2176. [PubMed] [Google Scholar]
  33. Kotb M., Ohnishi H., Majumdar G., Hackett S., Bryant A., Higgins G., Stevens D. Temporal relationship of cytokine release by peripheral blood mononuclear cells stimulated by the streptococcal superantigen pep M5. Infect Immun. 1993 Apr;61(4):1194–1201. doi: 10.1128/iai.61.4.1194-1201.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Lamche H. R., Adolf G. R. Highly sensitive enzyme immunoassays for antibodies to human tumor necrosis factor (TNF-alpha) and lymphotoxin (TNF-beta). J Immunol Methods. 1990 Aug 7;131(2):283–289. doi: 10.1016/0022-1759(90)90200-f. [DOI] [PubMed] [Google Scholar]
  35. Marrack P., Kappler J. The staphylococcal enterotoxins and their relatives. Science. 1990 May 11;248(4956):705–711. doi: 10.1126/science.2185544. [DOI] [PubMed] [Google Scholar]
  36. Matsuda T., Hirano T., Kishimoto T. Establishment of an interleukin 6 (IL 6)/B cell stimulatory factor 2-dependent cell line and preparation of anti-IL 6 monoclonal antibodies. Eur J Immunol. 1988 Jun;18(6):951–956. doi: 10.1002/eji.1830180618. [DOI] [PubMed] [Google Scholar]
  37. Mollick J. A., Cook R. G., Rich R. R. Class II MHC molecules are specific receptors for staphylococcus enterotoxin A. Science. 1989 May 19;244(4906):817–820. doi: 10.1126/science.2658055. [DOI] [PubMed] [Google Scholar]
  38. Mollick J. A., Miller G. G., Musser J. M., Cook R. G., Grossman D., Rich R. R. A novel superantigen isolated from pathogenic strains of Streptococcus pyogenes with aminoterminal homology to staphylococcal enterotoxins B and C. J Clin Invest. 1993 Aug;92(2):710–719. doi: 10.1172/JCI116641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Musser J. M., Hauser A. R., Kim M. H., Schlievert P. M., Nelson K., Selander R. K. Streptococcus pyogenes causing toxic-shock-like syndrome and other invasive diseases: clonal diversity and pyrogenic exotoxin expression. Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2668–2672. doi: 10.1073/pnas.88.7.2668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Norrby-Teglund A., Holm S. E., Norgren M. Detection and nucleotide sequence analysis of the speC gene in Swedish clinical group A streptococcal isolates. J Clin Microbiol. 1994 Mar;32(3):705–709. doi: 10.1128/jcm.32.3.705-709.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Paul N. L., Ruddle N. H. Lymphotoxin. Annu Rev Immunol. 1988;6:407–438. doi: 10.1146/annurev.iy.06.040188.002203. [DOI] [PubMed] [Google Scholar]
  42. Rordorf-Adam C., Lazdins J., Woods-Cook K., Alteri E., Henn R., Geiger T., Feige U., Towbin H., Erard F. An assay for the detection of interleukin-1 synthesis inhibitors: effects of antirheumatic drugs. Drugs Exp Clin Res. 1989;15(8):355–362. [PubMed] [Google Scholar]
  43. Rosenberg W. M., Moss P. A., Bell J. I. Variation in human T cell receptor V beta and J beta repertoire: analysis using anchor polymerase chain reaction. Eur J Immunol. 1992 Feb;22(2):541–549. doi: 10.1002/eji.1830220237. [DOI] [PubMed] [Google Scholar]
  44. Rosenfeld J., Capdevielle J., Guillemot J. C., Ferrara P. In-gel digestion of proteins for internal sequence analysis after one- or two-dimensional gel electrophoresis. Anal Biochem. 1992 May 15;203(1):173–179. doi: 10.1016/0003-2697(92)90061-b. [DOI] [PubMed] [Google Scholar]
  45. Sander B., Andersson J., Andersson U. Assessment of cytokines by immunofluorescence and the paraformaldehyde-saponin procedure. Immunol Rev. 1991 Feb;119:65–93. doi: 10.1111/j.1600-065x.1991.tb00578.x. [DOI] [PubMed] [Google Scholar]
  46. Schlievert P. M., Bettin K. M., Watson D. W. Purification and characterization of group A streptococcal pyrogenic exotoxin type C. Infect Immun. 1977 May;16(2):673–679. doi: 10.1128/iai.16.2.673-679.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Scholl P., Diez A., Mourad W., Parsonnet J., Geha R. S., Chatila T. Toxic shock syndrome toxin 1 binds to major histocompatibility complex class II molecules. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4210–4214. doi: 10.1073/pnas.86.11.4210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Stevens D. L., Tanner M. H., Winship J., Swarts R., Ries K. M., Schlievert P. M., Kaplan E. Severe group A streptococcal infections associated with a toxic shock-like syndrome and scarlet fever toxin A. N Engl J Med. 1989 Jul 6;321(1):1–7. doi: 10.1056/NEJM198907063210101. [DOI] [PubMed] [Google Scholar]
  49. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Uchiyama T., Imanishi K., Saito S., Araake M., Yan X. J., Fujikawa H., Igarashi H., Kato H., Obata F., Kashiwagi N. Activation of human T cells by toxic shock syndrome toxin-1: the toxin-binding structures expressed on human lymphoid cells acting as accessory cells are HLA class II molecules. Eur J Immunol. 1989 Oct;19(10):1803–1809. doi: 10.1002/eji.1830191007. [DOI] [PubMed] [Google Scholar]
  51. Uchiyama T., Kamagata Y., Yan X. J., Kohno M., Yoshioka M., Fujikawa H., Igarashi H., Okubo M., Awano F., Saito-Taki T. Study of the biological activities of toxic shock syndrome toxin-1: II. Induction of the proliferative response and the interleukin 2 production by T cells from human peripheral blood mononuclear cells stimulated with the toxin. Clin Exp Immunol. 1987 Jun;68(3):638–647. [PMC free article] [PubMed] [Google Scholar]
  52. WATSON D. W. Host-parasite factors in group A streptococcal infections. Pyrogenic and other effects of immunologic distinct exotoxins related to scarlet fever toxins. J Exp Med. 1960 Feb 1;111:255–284. doi: 10.1084/jem.111.2.255. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES