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The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1986 Nov 1;164(5):1700–1709. doi: 10.1084/jem.164.5.1700

Anthrax toxin blocks priming of neutrophils by lipopolysaccharide and by muramyl dipeptide

PMCID: PMC2188454  PMID: 3021891

Abstract

We studied the pretreatment of human polymorphonuclear neutrophils (PMN) with purified preparations of the anthrax toxin components-- protective antigen (PA), edema factor (EF), and lethal factor (LF)--and their effects on release of superoxide anion (O-2) after stimulation with the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (FMLP). PMN isolated in the absence of lipopolysaccharide (LPS) (less than 0.1 ng/ml) released only small amounts of O-2 after FMLP stimulation; pretreatment with anthrax toxin had little effect. The release of O-2 was increased fivefold by prior treatment with 3 ng/ml LPS for 1 h at 37 degrees C, an effect referred to as priming. PMN were primed to an equivalent extent by treatment with 100 ng/ml N-acetyl- muramyl-L-alanyl-D-isoglutamine (muramyl dipeptide [MDP]). Pretreatment of PMN with anthrax toxin components PA plus EF or PA plus LF inhibited priming by LPS or MDP, as shown by the reduction in the release of O-2 up to 90% relative to controls not treated with toxin; single toxin components were inactive. The inhibition was markedly reduced when priming with LPS or MDP was carried out before exposure to toxin. O-2 release after stimulation by phorbol myristate acetate was not increased by priming, and pretreatment with toxin did not inhibit O-2 release after this stimulus. Evidently, anthrax toxin inhibits the priming that is normally induced in PMN by bacterial products and is necessary for the full expression of antimicrobial effects.

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Selected References

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  1. Babior B. M. Oxidants from phagocytes: agents of defense and destruction. Blood. 1984 Nov;64(5):959–966. [PubMed] [Google Scholar]
  2. Babu U. M., Zeiger A. R. Soluble peptidoglycan from Staphylococcus aureus is a murine B-lymphocyte mitogen. Infect Immun. 1983 Dec;42(3):1013–1016. doi: 10.1128/iai.42.3.1013-1016.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chedid L., Audibert F., Johnson A. G. Biological activities of muramyl dipeptide, a synthetic glycopeptide analogous to bacterial immunoregulating agents. Prog Allergy. 1978;25:63–105. [PubMed] [Google Scholar]
  4. Ezzell J. W., Ivins B. E., Leppla S. H. Immunoelectrophoretic analysis, toxicity, and kinetics of in vitro production of the protective antigen and lethal factor components of Bacillus anthracis toxin. Infect Immun. 1984 Sep;45(3):761–767. doi: 10.1128/iai.45.3.761-767.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gaffin S. L., Badsha N., Brock-Utne J. G., Vorster B. J., Conradie J. D. An ELISA procedure for detecting human anti-endotoxin antibodies in serum. Ann Clin Biochem. 1982 May;19(3):191–194. doi: 10.1177/000456328201900313. [DOI] [PubMed] [Google Scholar]
  6. Gallin J. I., Wright D. G., Schiffmann E. Role of secretory events in modulating human neutrophil chemotaxis. J Clin Invest. 1978 Dec;62(6):1364–1374. doi: 10.1172/JCI109257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Guthrie L. A., McPhail L. C., Henson P. M., Johnston R. B., Jr Priming of neutrophils for enhanced release of oxygen metabolites by bacterial lipopolysaccharide. Evidence for increased activity of the superoxide-producing enzyme. J Exp Med. 1984 Dec 1;160(6):1656–1671. doi: 10.1084/jem.160.6.1656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Haslett C., Guthrie L. A., Kopaniak M. M., Johnston R. B., Jr, Henson P. M. Modulation of multiple neutrophil functions by preparative methods or trace concentrations of bacterial lipopolysaccharide. Am J Pathol. 1985 Apr;119(1):101–110. [PMC free article] [PubMed] [Google Scholar]
  9. Issekutz A. C., Ng M., Biggar W. D. Effect of cyclic adenosine 3',5'-monophosphate antagonists on endotoxin-induced inhibition of human neutrophil chemotaxis. Infect Immun. 1979 May;24(2):434–440. doi: 10.1128/iai.24.2.434-440.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Johnston R. B., Jr, Kitagawa S. Molecular basis for the enhanced respiratory burst of activated macrophages. Fed Proc. 1985 Nov;44(14):2927–2932. [PubMed] [Google Scholar]
  11. KEPPIE J., HARRIS-SMITH P. W., SMITH H. THE CHEMICAL BASIS OF THE VIRULENCE OF BACILLUS ANTHRACIS. IX. ITS AGGRESSINS AND THEIR MODE OF ACTION. Br J Exp Pathol. 1963 Aug;44:446–453. [PMC free article] [PubMed] [Google Scholar]
  12. Kaku M., Yagawa K., Nagao S., Tanaka A. Enhanced superoxide anion release from phagocytes by muramyl dipeptide or lipopolysaccharide. Infect Immun. 1983 Feb;39(2):559–564. doi: 10.1128/iai.39.2.559-564.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leppla S. H. Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells. Proc Natl Acad Sci U S A. 1982 May;79(10):3162–3166. doi: 10.1073/pnas.79.10.3162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Leppla S. H. Bacillus anthracis calmodulin-dependent adenylate cyclase: chemical and enzymatic properties and interactions with eucaryotic cells. Adv Cyclic Nucleotide Protein Phosphorylation Res. 1984;17:189–198. [PubMed] [Google Scholar]
  15. Lincoln R. E., Walker J. S., Klein F., Rosenwald A. J., Jones W. I., Jr Value of field data for extrapolation in anthrax. Fed Proc. 1967 Sep;26(5):1558–1562. [PubMed] [Google Scholar]
  16. MARGOLIASH E., FROHWIRT N. Spectrum of horse-heart cytochrome c. Biochem J. 1959 Mar;71(3):570–572. doi: 10.1042/bj0710570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nishizuka Y. Turnover of inositol phospholipids and signal transduction. Science. 1984 Sep 21;225(4668):1365–1370. doi: 10.1126/science.6147898. [DOI] [PubMed] [Google Scholar]
  18. O'Brien J., Friedlander A., Dreier T., Ezzell J., Leppla S. Effects of anthrax toxin components on human neutrophils. Infect Immun. 1985 Jan;47(1):306–310. doi: 10.1128/iai.47.1.306-310.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. PUZISS M., WRIGHT G. G. Studies on immunity in anthrax. X. Gel-adsorbed protective antigen for immunization of man. J Bacteriol. 1963 Jan;85:230–236. doi: 10.1128/jb.85.1.230-236.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. SMITH H., KEPPIE J., STANLEY J. L. The chemical basis of the virulence of Bacillus anthracis. V. The specific toxin produced by B. Anthracis in vivo. Br J Exp Pathol. 1955 Oct;36(5):460–472. [PMC free article] [PubMed] [Google Scholar]
  21. STANLEY J. L., SMITH H. The three factors of anthrax toxin: their immunogenicity and lack of demonstrable enzymic activity. J Gen Microbiol. 1963 May;31:329–337. doi: 10.1099/00221287-31-2-329. [DOI] [PubMed] [Google Scholar]
  22. Seligmann B., Malech H. L., Melnick D. A., Gallin J. I. An antibody binding to human neutrophils demonstrates antigenic heterogeneity detected early in myeloid maturation which correlates with functional heterogeneity of mature neutrophils. J Immunol. 1985 Oct;135(4):2647–2653. [PubMed] [Google Scholar]
  23. Smedly L. A., Tonnesen M. G., Sandhaus R. A., Haslett C., Guthrie L. A., Johnston R. B., Jr, Henson P. M., Worthen G. S. Neutrophil-mediated injury to endothelial cells. Enhancement by endotoxin and essential role of neutrophil elastase. J Clin Invest. 1986 Apr;77(4):1233–1243. doi: 10.1172/JCI112426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Spitznagel J. K., Shafer W. M. Neutrophil killing of bacteria by oxygen-independent mechanisms: a historical summary. Rev Infect Dis. 1985 May-Jun;7(3):398–403. doi: 10.1093/clinids/7.3.398. [DOI] [PubMed] [Google Scholar]
  25. Stephen J. Anthrax toxin. Pharmacol Ther. 1981;12(3):501–513. doi: 10.1016/0163-7258(81)90095-4. [DOI] [PubMed] [Google Scholar]
  26. Vacheron F., Guenounou M., Nauciel C. Induction of interleukin 1 secretion by adjuvant-active peptidoglycans. Infect Immun. 1983 Dec;42(3):1049–1054. doi: 10.1128/iai.42.3.1049-1054.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wade B. H., Wright G. G., Hewlett E. L., Leppla S. H., Mandell G. L. Anthrax toxin components stimulate chemotaxis of human polymorphonuclear neutrophils. Proc Soc Exp Biol Med. 1985 Jun;179(2):159–162. doi: 10.3181/00379727-179-42078. [DOI] [PubMed] [Google Scholar]

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