Anthrax Pathogenesis and Host Response

  • P. Hanna
Part of the Current Topics in Microbiology and Immunology book series (CT MICROBIOLOGY, volume 225)


Anthrax has been both a scourge and a fundamental model for infectious disease studies for over a century. Death associated with systemic anthrax is mimicked in animals challenged with anthrax lethal toxin, a virulence factor believed to affect only macrophages. Animals depleted of macrophages become resistant to the toxin, while reintroduction of cultured macrophages into depleted animals restores sensitivity. These studies and others implicate an active role for the innate immune system in the demise of the anthrax victim. Many of the molecular factors and events in the cascade of lethal events during anthrax infections have now been identified. Other recent overviews of anthrax pathogenesis and toxins include those by Stephen (1986), Friedlander (1990), Leppla (1995), and Hanna and Collier (1997).


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  1. Anonymous (1994) Anthrax control and research, with special reference to national programme development in Africa: memorandum from a WHO meeting. Bull World Health Organ 72:13–22Google Scholar
  2. Arora N, Leppla SH (1993) Residues 1–254 of anthrax toxin lethal factor are sufficient to cause cellular uptake of fused polypeptides. J Biol Chem 268:3334–3341PubMedGoogle Scholar
  3. Arora N, Leppla SH (1994) Fusions of anthrax toxin lethal factor with shiga toxin and diphtheria toxin enzymatic domains are toxic to mammalian cells. Infect Immun 62:4955–4961PubMedGoogle Scholar
  4. Arora N, Klimpel KR, Singh Y, Leppla SH (1992) Fusions of anthrax toxin lethal factor to the ADP-ribosylation domain of Pseudomonas exotoxin A are potent cytotoxins which are translocated to the cytosol of mammalian cells. J Biol Chem 267:15542–15548PubMedGoogle Scholar
  5. Bartkus JM, Leppla SH (1989) Transcriptional regulation of the protective antigen gene of Bacillus anthracis. Infect Immun 57:22295–22299Google Scholar
  6. Bell JH (1880) On anthrax and anthracaemia in wool sorters, heifers and sheep. Br Med J 2:656–657PubMedGoogle Scholar
  7. Blaustein RO, Koehler TM, Collier RJ, Finkelstein A (1989) Anthrax toxin: channel-forming activity of protective antigen in planar phospholipid bilayers. Proc Natl Acad Sci USA 86:2209–2213PubMedGoogle Scholar
  8. Blaustein RO, Lea EJ, Finkelstein A (1990) Voltage-dependent block of anthrax toxin channels in planar phospholipid bilayer membranes by symmetric tetraalkylammonium ions: single-channel analysis. J Gen Physiol 96:921–942PubMedGoogle Scholar
  9. Bragg TS, Robertson DL (1989) Nucleotide sequence and analysis of the lethal factor gene (lef) from Bacillus anthracis. Gene 81:45–54PubMedGoogle Scholar
  10. Cataldi A, Labruyere E, Mock M (1990) Construction and characterization of a protective antigen-deficient Bacillus anthracis strain. Mol Microbiol 4:1111–1117PubMedGoogle Scholar
  11. Cataldi A, Fouet A, Mock M (1992) Regulation of pag gene expression in Bacillus anthracis: use of a paglacZ transcriptional fusion. FEMS Microbiol Lett 98:89–93Google Scholar
  12. Confer DL, Eaton JW (1982) Phagocyte impotence caused by the invasive bacterial adenylate cyclase. Science 217:948–950PubMedGoogle Scholar
  13. Dai Z, Sirard JC, Mock M, Koehler TM (1995) The atxA gene product activates transcription of the anthrax toxin genes and is essential for virulence. Mol Microbiol 16:1171–1181PubMedGoogle Scholar
  14. Dinarello CA (1988) Biology of interleukin 1. FASEB J 2:108–115PubMedGoogle Scholar
  15. Dirckx JH (1981) Virgil on anthrax. Am J Dermatopathol 3:191–195PubMedGoogle Scholar
  16. Escuyer V, Collier RJ (1991) Anthrax protective antigen interacts with a specific receptor on the surface of CHO-K1 cells. Infect Immun 59:3381–3386PubMedGoogle Scholar
  17. Escuyer V, Duflot E, Sezer O, Danchin A, Mock M (1988) Structural homology between virulence-associated bacterial adenylate cyclases. Gene 71:293–298PubMedGoogle Scholar
  18. Ezekowitz RAB (1992) Chronic granulomatous disease: an update and a paradigm for the use of interferon-gamma as adjunct immunotherapy in infectious diseases. Curr Topics Microbiol Immunol 181:283–292Google Scholar
  19. Fawthrop DJ, Boobis AR, Davies DS (1991) Mechanisms of cell death. Arch Toxicol 65:437–444PubMedGoogle Scholar
  20. Friedlander AM (1986) Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process. J Biol Chem 261:7123–7126PubMedGoogle Scholar
  21. Friedlander AM (1990) The anthrax toxins. In: Saelinger CB (ed) Trafficking of bacterial toxins. CRC Press, Boca Raton, pp 121–138Google Scholar
  22. Friedlander A, Bhatnagar R, Leppla SH, Johnson L, Singh Y (1993) Characterization of macrophage sensitivity and resistance to anthrax lethal toxin. Infect Immun 61:245–252PubMedGoogle Scholar
  23. Gill DM (1978) Seven toxin peptides that cross cell membranes. In: Jeljaszewicz J, Wadstrom T (eds) Bacterial toxins and cell membranes. Academic, New York, pp 291–332Google Scholar
  24. Gladstone GP (1946) Immunity to anthrax. Protective antigen present in cell-free culture filtrates. Br J Exp Pathol 27:349–418Google Scholar
  25. Gordon VM, Leppla SH, Hewlett EL (1988)Inhibitors of receptor-mediated endocytosis block the entry of Bacillus anthracis adenylate cyclase toxin but not that of Bordetella pertussis adenylate cyclase toxin. Infect Immun 56:1066–1069PubMedGoogle Scholar
  26. Gordon VM, Young WW, Lechler SM, Gray Mc, Leppla SH, Hewlett EL (1989) Adenylate cyclase toxins from Bacillus anthracis and Bordetella pertussis. Different processes for interaction with and entry into target cells. J Biol Chem 264:14792–14796PubMedGoogle Scholar
  27. Hanna PC, Collier RJ (1997) Anthrax lethal toxin. In: Rapoulli R (ed) Bacterial toxins and their uses in cell biology (in press)Google Scholar
  28. Hanna PC, Kochi S, Collier RJ (1992) Biochemical and physiological changes induced by anthrax lethal toxin in J774 macrophage-like cells. Mol Biol Cell 3:1269–1277PubMedGoogle Scholar
  29. Hanna PC, Acosta D, Collier R (1993) On the role of macrophages in anthrax. Proc Natl Acad Sci USA 90:10198–10201PubMedGoogle Scholar
  30. Hanna PC, Kruskal B, Ezekowitz R, Bloom B, Collier RJ (1994) Role of macrophages oxidative burst in the action of anthrax lethal toxin. Mol Med 1:7–18PubMedGoogle Scholar
  31. Hoover DL, Friedlander AM, Rogers LC, Yoon IK, Warren RL, Cross AS (1994) Anthrax edema toxin differentially regulates lipopolysaccharide-induced monocyte production of tumor necrosis factor alpha and interleukin-6 by increasing intracellular cyclic AMP. Infect Immun 62:4432–4439PubMedGoogle Scholar
  32. Inocencio NM, Moehring JM, Moehring TJ (1993) A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A is unable to process the fusion glycoprotein of Newcastle disease virus. J Virol 67:595Google Scholar
  33. Kagan E, Hartman D (1984) Specific depletion of macrophages by silica treatment. Methods Enzymol 108:325–335PubMedGoogle Scholar
  34. Kaspar RL, Robertson DL (1987) Purification and physical analysis of Bacillus anthracis plasmids pXOl and pXO2. Biochem Biophys Res Commun 149:362–368PubMedGoogle Scholar
  35. Kass EH, Kendrick MI, Tsai YC, Parsonnet J (1987) Interaction of magnesium ion, oxygen tension and temperature in the production of toxic shock syndrome toxin-1 by Staphylococcus aureus. J Infect Dis 155:812–815PubMedGoogle Scholar
  36. Klimpel KR, Molloy SS, Thomas G, Leppla SH (1992) Anthrax toxin protective antigen is activated by a cell surface protease with the sequence specificity and catalytic properties of furin. Proc Natl Acad Sci USA 89:10277–10281PubMedGoogle Scholar
  37. Klimpel KR, Arora N, Leppla SH (1993) Anthrax toxin lethal factor has homology to the thermolysin-like proteases and displays proteolytic activity. Ann Meet Am Soc Microbiol 45:B–111Google Scholar
  38. Klimpel KR, Arora N, Leppla SH (1994) Anthrax toxin lethal factor contains a zinc metalloprotease consensus sequence which is required for lethal toxin activity. Mol Microbiol 13:1093–1100PubMedGoogle Scholar
  39. Koch R (1877) The aetiology of anthrax based on the ontogeny of the anthrax bacillus. Beitr Biol Pflanz 2:277–282Google Scholar
  40. Kochi SK, Schiavo G, Mock M, Montecucco C (1994) Zinc content of the Bacillus anthracis lethal factor. FEMS Microbiol Lett 124:343–348PubMedGoogle Scholar
  41. Koehler TM, Collier RJ (1991) Anthrax toxin protective antigen: low pH-induced hydrophobicity and channel formation in liposomes. Mol Microbiol 5:1501–1506PubMedGoogle Scholar
  42. Koehler TM, Dai Z, Kaufman-Yarbray M (1994) Regulation of the Bacillus anthracis protective antigen gene: CO2 and a trans-acting element activate transcription from one of two prometers. J Bacteriol 176:586–595PubMedGoogle Scholar
  43. Labruyere E, Mock M, Ladant D, Michelson S, Gilles AM, Laoide B, Baarzu O (1990) Characterization of ATP and calmodulin-binding properties of a truncated form of Bacillus anthracis adenylate cyclase. Biochemistry 29:4922–4928PubMedGoogle Scholar
  44. LaForce FM (1978) Woolsorter’s disease in England. NY Acad Med 54:956Google Scholar
  45. Lenardo M J, Baltimore D (1989) NF-kB: a pleiotropic mediator of inducable and tissue-specific gene control. Cell 58:227–229PubMedGoogle Scholar
  46. Leppla SH (1982) Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations in eukaryotic cells. Proc Natl Acad Sci USA 79:3162–3166PubMedGoogle Scholar
  47. Leppla SH (1984) Bacillus anthracis calmodulin-dependent adenylate cyclase: chemical and enzymatic properties and interactions with eukaryotic cells. Adv Cyclic Nuleotide Protein Phosphorylat Res 17:189–198Google Scholar
  48. Leppla SH (1988) Production and purification of anthrax toxin. Methods Enzymol 165:103–116PubMedGoogle Scholar
  49. Leppla SH (1991) The anthrax toxin complex. In: Alouf JE, Freer JH (eds) Sourcebook of bacterial protein toxins. Academic, London, pp 277–301Google Scholar
  50. Leppla SH (1995) Anthrax toxins. In: Moss J, Iglewski B, Vaughan M, Tu AT (eds) Bacterial toxins and virulence factors in disease. Dekker, New York, pp 543–572Google Scholar
  51. Leppla SH, Ivins BE, Ezzell JW (1985) Anthrax toxin. In: Leive L, Bonventre PF, Morello JA, Schlessinger S, Silver SD, Wu HC (eds) Microbiology. American Society of Microbiology, Washington DC, pp 63–66Google Scholar
  52. Lincoln RE, Fish DC (1970) Anthrax toxin. In: Montie TC, Kadis S, Ajl SJ (eds) Microbial toxins, vol III. Academic, New York, pp 361–414Google Scholar
  53. Little SF, Lowe JR (1991) Location of the receptor-binding region of protective antigen from Bacillus anthracis. Biochem Biophys Res Commun 180:531–537PubMedGoogle Scholar
  54. Meselson M, Guillemin J, Hugh-Jones M, Langmuir A, Popova I, Shelokov A, Yampolskaya O (1994) The Sverdlovsk anthrax outbreak of 1979. Science 266:1202–1208PubMedGoogle Scholar
  55. Metchnikoff E (1905) Immunity in infective diseases. Cambridge University Press, LondonGoogle Scholar
  56. Milne JC, Collier RJ (1993) pH-dependent permeabilization of the plasma membrane of mammalian cells by anthrax protective antigen. Mol Microbiol 10:647–653PubMedGoogle Scholar
  57. Milne JC, Furlong D, Hanna PC, Wall JS, Collier RJ (1994) Anthrax protective antigen forms oligomers during intoxication of mammalian cells. J Biol Chem 269:20607PubMedGoogle Scholar
  58. Milne JC, Blanke SR, Hanna PC, Collier RJ (1995) Protective antigen-binding domain of anthrax lethal factor mediates translocation of a heterologous protein fused to its amino- or carboxy-terminus. Mol Microbiol 15:661–666PubMedGoogle Scholar
  59. Mock M, Labruyere E, Glaser P, Danchin A, Ullmann A (1988) Cloning and expression of the calmodulin-sensitive Bacillus anthracis adenylate cyclase in Escherichia coli. Gene 64:277–284PubMedGoogle Scholar
  60. Molloy SS, Bresnahan PA, Leppla SH, Klimpel KR, Thomas G (1992) Humin furin is a calcium-dependent serine endoprotease that recognizes the sequence Arg-X-X-Arg and efficiently cleaves anthrax toxin protective antigen. J Biol Chem 267:16396–16402PubMedGoogle Scholar
  61. Montecucco C, Schiavo G (1993) Tetanus and botulism neurotoxins: a new group of zinc proteases. Trend Biochem Sci 18:324–327PubMedGoogle Scholar
  62. Munier H, Blanco FJ, Prêcheur B, Diesis E, Nieto JL, Craescu CT, Barzu O (1993) Characterization of a synthetic calmodulin-binding peptide derived from Bacillus anthracis adenylate cyclase. J Biol Chem 268:1695–1701PubMedGoogle Scholar
  63. Nakayama S, Kretsinger RH (1994) Evolution of the EF-hand family of proteins. Annu Rev Biophys Biomol Struct 23:473–475PubMedGoogle Scholar
  64. Novak JM, Stein MP, Little SF, Leppla SH, Friedlander AM (1992) Functional characterization of protease-treated Bacillus anthracis protective antigen. J Biol Chem 267:17186–17193PubMedGoogle Scholar
  65. O’Brien J, Friedlander A, Dreier T, Ezzell J, Leppla S (1985) Effects of anthrax toxin components on human neutrophils. Infect Immun 47:306–310PubMedGoogle Scholar
  66. Ogata M, Fryling CM, Pastan I, FitzGerald DJ (1992) Cell-mediated cleavage of Pseudomonas exotoxin between ARG279 and Gly280 generates the enzymatically active fragment which translocates to the cytosol. J Biol Chem 267:25396–25401PubMedGoogle Scholar
  67. Pasteur L (1881) De l’atténuation des virus et de leur retour a la virulence. CR Acad Sci Agric Bulg 92:429–435Google Scholar
  68. Perelle S, Gibert M, Boquet P, Popoff MR (1993) Characterization of Clostridium perfringens iota-toxin genes and expression in Escherichia coli. Infect Immun 61:5147–5156PubMedGoogle Scholar
  69. Petosa C, Liddington RC (1997) The anthrax toxin. In: Parker M W (ed) Protein toxin structure. Landes, Austin (in press)Google Scholar
  70. Petosa C, Collier RJ, Klimpel KR, Leppla SH, Liddington RC (1997) Crystal structure of the anthrax toxin protective antigen. Nature 385:833–838PubMedGoogle Scholar
  71. Pezard C, Berche P, Mock M (1991) Contribution of individual toxin components to virulence of Bacillus anthracis. Infect Immun 59:3472–3477PubMedGoogle Scholar
  72. Pezard C, Duflot E, Mock M (1993) Construction of Bacillus anthracis mutant strains producing a single toxin component. J Gen Microbiol 139:2459–2463PubMedGoogle Scholar
  73. Quinn CP, Singh Y, Klimpel KR, Leppla SH (1991) Functional mapping of anthrax toxin lethal factor by in-frame insertion mutagenesis. J Biol Chem 266:20124–20130PubMedGoogle Scholar
  74. Robertson DL (1988) Relationships between the calmodulin-dependent adenylate cyclases produced by Bacillus anthracis and Bordetella pertussis. Biochem Biophys Res Commun 157:1027–1032PubMedGoogle Scholar
  75. Robertson DL, Leppla SH (1986) Molecular cloning and expression in Escherichia coli of the lethal factor gene of Bacillus anthracis. Gene 44:71–78PubMedGoogle Scholar
  76. Robertson DL, Tippetts MT, Leppla SH (1988) Nucleotide sequence of the Bacillus anthracis edema factor gene (cya): a calmodulin-dependent adenylate cyclase. Gene 73:363–371PubMedGoogle Scholar
  77. Robertson DL, Bragg TS, Simpson S, Kaspar R, Xie W, Tippetts MT (1990) Mapping and characterization of the of Bacillus anthracis plasmids pXOl and pX02. Salisbury Med Bull 68:55–58Google Scholar
  78. Schiavo G, Benfenati F, Poulain B, Rossetto O, de Laureto PP, Dasgupta BR, Montecucco C (1992) Tetanus and botilinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin. Nature 359:832–835PubMedGoogle Scholar
  79. Schreck R, Rieber P, Baeuerle PA (1991) Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-kB transcription factor and HIV-1. EM BO J 10:2247–2258Google Scholar
  80. Shimamura T, Watanabe S, Sasaki S (1985) Enhancement of enterotoxin production by carbon dioxide in Vibrio cholerae. Infect Immun 49:455–456PubMedGoogle Scholar
  81. Singh Y, Chaudhary VK, Leppla SH (1989) A deleted varient of Bacillus anthracis protective antigen is non-toxic and blocks anthrax toxin in vivo. J Biol Chem 264:19103–19107PubMedGoogle Scholar
  82. Singh Y, Klimpel KR, Quinn CP, Chaudhary VK, Leppla SH (1991) The carboxyl-terminal end of protective antigen is required for receptor-binding and anthrax toxin activity. J Biol Chem 266:15493–15497PubMedGoogle Scholar
  83. Singh Y, Klimpel KR, Arora N, Sharma M, Leppla SH (1994) The chymotrypsin-sensitive site FFD315, in anthrax toxin protective antigen is required for translocation of lethal factor. J Biol Chem 269:29039–29046PubMedGoogle Scholar
  84. Sirarad JC, Mock M, Fouet A (1994) The three Bacillus anthracis toxin genes are coordinately regulated by bicarbonate and temperature. J Bacteriol 176:5188–5192Google Scholar
  85. Steiner DF, Smeekens SP, Ohagi S, Chan SJ (1992) The new enzymology of precursor processing endoproteases. J Biol Chem 267:23435–23438PubMedGoogle Scholar
  86. Stephen J (1986) Anthrax toxin. In: Dorner F, Drews J (eds) Pharmacology of bacterial toxins. Pergamon, Oxford, pp 381–395Google Scholar
  87. Südhof TC, De Camilli P, Niemann H, Jahn R (1993) Membrane fusion machinery: insights from synaptic proteins. Cell 75:1–4PubMedGoogle Scholar
  88. Turnbull PC (1992) Anthrax vaccines: past, present and future. Vaccine 9:533—539Google Scholar
  89. Uchida I, Sekizaki T, Hashimoto K, Terakado N (1985) Association of the encapsulation of Bacillus anthracis with a 60-megadalton plasmid. J Gen Microbiol 131:363–3367PubMedGoogle Scholar
  90. Uchida I, Hashimoto K, Makino S, Sasakawa C, Yoshikawa M, Teradado N (1987) Restriction map of a capsule plasmid of Bacillus anthracis. Plasmid 18:178–181PubMedGoogle Scholar
  91. Uchida I, Hornung JM, Thorne CB, Klimpel KR, Leppla SH (1993) Cloning and characterization of a gene whose product is a transactivator of anthrax toxin synthesis. J Bacteriol 175:5329–5338PubMedGoogle Scholar
  92. Vallée BL, Auld DS (1990) Zinc coordination, function, and structures of zinc enzymes and other proteins. Biochemistry 29:5647–5659PubMedGoogle Scholar
  93. van de Ven WJ, Voorberg J, Fontign R, Pannekoek H, van den Ouweland AM, van Duijnhoven HL, Roebroek AJ, and Siezen RJ (1990) Furin is a subtilisn-like proprotein-processing enzyme in higher eukaryotes. Mol Biol Rep 14:265–275PubMedGoogle Scholar
  94. Vietri NJ, Marrero R,. Hoover TA, Welkos SL (1995) Indentification and characterization of a transactivator involved in the regulation of encapsulation by Bacillus anthracis. Gene 152:1–9PubMedGoogle Scholar
  95. Vodkin MH, Leppla SH (1983) Cloning of the protective antigen gene of Bacillus anthracis. Cell 34:693–697PubMedGoogle Scholar
  96. Wade B, Wright G, Hewlett E, Leppla S, Mandell G (1985) Anthrax toxin components stimulate Chemotaxis of human polymorphonuclear neutrophils. Proc Soc Exp Biol Med 179:159–162PubMedGoogle Scholar
  97. Walker B, Braha O, Cheley S, Bayley H (1995) An intermediate in the assembly of a pore-forming protein trapped with a genetically-engineered switch. Chem Biol 2:99–105PubMedGoogle Scholar
  98. Warren G (1996) Novel pesticidal proteins and strains. World intellectual property organization. Patent application WO 96/10083Google Scholar
  99. Welkos SL (1991) Plasmid-associated virulence factors of non-toxigenic (pXOl-) Bacillus anthracis. Microb Pathogen 10:183–198Google Scholar
  100. Welkos SL, Keener TJ, Gibbs PH (1986) Differences in susceptibility of inbred mice to Bacillus anthracis. Infect Immun 51:795–800PubMedGoogle Scholar
  101. Welkos S, Lowe J, Eden-McCutchan F, Vodkin M, Leppla S, Schmidt J (1988) Sequence and analysis of the DNA excoding protective antigen of Bacillus anthracis. Gene 69:287–300PubMedGoogle Scholar
  102. Williams DP, Wen Z, Watson RS, Boyd J, Strom TB, Murphy JR (1990) Cellular processing of the interleukin-2 fusion toxin DAB486-IL-2 and efficient delivery of diphtheria fragment A to the cytosol of target cells requires Arg 194. J Biol Chem 265:20673–20677PubMedGoogle Scholar
  103. Wright GG, Mandell GL (1986) Anthrax toxin blocks priming of neutrophils by lipopolysaccharide and by muramyl dipeptide. J Exp Med 164:1700–1709PubMedGoogle Scholar
  104. Wright GG, Read PW, Mandell GL (1988) Lipopolysaccharide releases a priming substance from platelets that augments the oxidative response of polymorphonuclear neutrophils to chemotactic peptide. J Infect Dis 157:690–696PubMedGoogle Scholar
  105. Xia Z, Storm DR (1990) A-type ATP-binding consensus sequences are critical for the catalytic activity of the calmodulin-sensitive adenyly cyclase from Bacillus anthracis. J Biol Chem 265:6517–6520PubMedGoogle Scholar

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© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • P. Hanna
    • 1
  1. 1.Department of Microbiology, Department of ImmunologyDuke University Medical CenterDurhamUSA

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