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. 1995 Apr;63(4):1369–1372. doi: 10.1128/iai.63.4.1369-1372.1995

Protective immunity induced by Bacillus anthracis toxin-deficient strains.

C Pezard 1, M Weber 1, J C Sirard 1, P Berche 1, M Mock 1
PMCID: PMC173160  PMID: 7890396

Abstract

The two toxins secreted by Bacillus anthracis are composed of binary combinations of three proteins: protective antigen (PA), lethal factor (LF), and edema factor (EF). Six mutant strains that are deficient in the production of one or two of these toxin components have been previously constructed and characterized (C. Pezard, E. Duflot, and M. Mock, J. Gen. Microbiol. 139:2459-2463, 1993). In this work, we examined the antibody response to the in vivo production of PA, LF, and EF in mice immunized with spores of strains producing these proteins. High titers of antibody to PA were observed after immunization with all strains producing PA, while titers of antibodies to EF and LF were weak in animals immunized with strains producing only EF or LF. In contrast, immunization with strains producing either PA and EF or PA and LF resulted in an increased antibody response to EF or LF, respectively. The differing levels of protection from a lethal anthrax challenge afforded to mice immunized with spores of the mutant strains not only confirm the role of PA as the major protective antigen in the humoral response but also indicate a significant contribution of LF and EF to immunoprotection. We observed, however, that PA-deficient strains were also able to provide some protection, thereby suggesting that immune mechanisms other than the humoral response may be involved in immunity to anthrax. Finally, a control strain lacking the toxin-encoding plasmid was unable to provide protection or elicit an antibody response against bacterial antigens, indicating a possible role for pXO1 in the survival of B. anthracis in a host.

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Selected References

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  1. BEALL F. A., TAYLOR M. J., THORNE C. B. Rapid lethal effect in rats of a third component found upon fractionating the toxin of Bacillus anthracis. J Bacteriol. 1962 Jun;83:1274–1280. doi: 10.1128/jb.83.6.1274-1280.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cataldi A., Labruyère E., Mock M. Construction and characterization of a protective antigen-deficient Bacillus anthracis strain. Mol Microbiol. 1990 Jul;4(7):1111–1117. doi: 10.1111/j.1365-2958.1990.tb00685.x. [DOI] [PubMed] [Google Scholar]
  3. Fish D. C., Mahlandt B. G., Dobbs J. P., Lincoln R. E. Purification and properties of in vitro-produced anthrax toxin components. J Bacteriol. 1968 Mar;95(3):907–918. doi: 10.1128/jb.95.3.907-918.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Green B. D., Battisti L., Koehler T. M., Thorne C. B., Ivins B. E. Demonstration of a capsule plasmid in Bacillus anthracis. Infect Immun. 1985 Aug;49(2):291–297. doi: 10.1128/iai.49.2.291-297.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hambleton P., Carman J. A., Melling J. Anthrax: the disease in relation to vaccines. Vaccine. 1984 Jun;2(2):125–132. doi: 10.1016/0264-410x(84)90003-3. [DOI] [PubMed] [Google Scholar]
  6. Hanna P. C., Acosta D., Collier R. J. On the role of macrophages in anthrax. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10198–10201. doi: 10.1073/pnas.90.21.10198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ivins B. E., Ezzell J. W., Jr, Jemski J., Hedlund K. W., Ristroph J. D., Leppla S. H. Immunization studies with attenuated strains of Bacillus anthracis. Infect Immun. 1986 May;52(2):454–458. doi: 10.1128/iai.52.2.454-458.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ivins B. E., Welkos S. L., Knudson G. B., Little S. F. Immunization against anthrax with aromatic compound-dependent (Aro-) mutants of Bacillus anthracis and with recombinant strains of Bacillus subtilis that produce anthrax protective antigen. Infect Immun. 1990 Feb;58(2):303–308. doi: 10.1128/iai.58.2.303-308.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ivins B. E., Welkos S. L., Little S. F., Crumrine M. H., Nelson G. O. Immunization against anthrax with Bacillus anthracis protective antigen combined with adjuvants. Infect Immun. 1992 Feb;60(2):662–668. doi: 10.1128/iai.60.2.662-668.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Klimpel K. R., Arora N., Leppla S. H. Anthrax toxin lethal factor contains a zinc metalloprotease consensus sequence which is required for lethal toxin activity. Mol Microbiol. 1994 Sep;13(6):1093–1100. doi: 10.1111/j.1365-2958.1994.tb00500.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Little S. F., Knudson G. B. Comparative efficacy of Bacillus anthracis live spore vaccine and protective antigen vaccine against anthrax in the guinea pig. Infect Immun. 1986 May;52(2):509–512. doi: 10.1128/iai.52.2.509-512.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mahlandt B. G., Klein F., Lincoln R. E., Haines B. W., Jones W. I., Jr, Friedman R. H. Immunologic studies of anthrax. IV. Evaluation of the immunogenicity of three components of anthrax toxin. J Immunol. 1966 Apr;96(4):727–733. doi: 10.21236/ad0640357. [DOI] [PubMed] [Google Scholar]
  14. Makino S., Sasakawa C., Uchida I., Terakado N., Yoshikawa M. Cloning and CO2-dependent expression of the genetic region for encapsulation from Bacillus anthracis. Mol Microbiol. 1988 May;2(3):371–376. doi: 10.1111/j.1365-2958.1988.tb00041.x. [DOI] [PubMed] [Google Scholar]
  15. Mikesell P., Ivins B. E., Ristroph J. D., Dreier T. M. Evidence for plasmid-mediated toxin production in Bacillus anthracis. Infect Immun. 1983 Jan;39(1):371–376. doi: 10.1128/iai.39.1.371-376.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mock M., Ullmann A. Calmodulin-activated bacterial adenylate cyclases as virulence factors. Trends Microbiol. 1993 Aug;1(5):187–192. doi: 10.1016/0966-842x(93)90089-a. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Pezard C., Berche P., Mock M. Contribution of individual toxin components to virulence of Bacillus anthracis. Infect Immun. 1991 Oct;59(10):3472–3477. doi: 10.1128/iai.59.10.3472-3477.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pezard C., Duflot E., Mock M. Construction of Bacillus anthracis mutant strains producing a single toxin component. J Gen Microbiol. 1993 Oct;139(10):2459–2463. doi: 10.1099/00221287-139-10-2459. [DOI] [PubMed] [Google Scholar]
  20. STANLEY J. L., SMITH H. Purification of factor I and recognition of a third factor of the anthrax toxin. J Gen Microbiol. 1961 Sep;26:49–63. doi: 10.1099/00221287-26-1-49. [DOI] [PubMed] [Google Scholar]
  21. Sirard J. C., Mock M., Fouet A. The three Bacillus anthracis toxin genes are coordinately regulated by bicarbonate and temperature. J Bacteriol. 1994 Aug;176(16):5188–5192. doi: 10.1128/jb.176.16.5188-5192.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Turnbull P. C., Broster M. G., Carman J. A., Manchee R. J., Melling J. Development of antibodies to protective antigen and lethal factor components of anthrax toxin in humans and guinea pigs and their relevance to protective immunity. Infect Immun. 1986 May;52(2):356–363. doi: 10.1128/iai.52.2.356-363.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Uchida I., Hornung J. M., Thorne C. B., Klimpel K. R., Leppla S. H. Cloning and characterization of a gene whose product is a trans-activator of anthrax toxin synthesis. J Bacteriol. 1993 Sep;175(17):5329–5338. doi: 10.1128/jb.175.17.5329-5338.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Welkos S. L., Friedlander A. M. Comparative safety and efficacy against Bacillus anthracis of protective antigen and live vaccines in mice. Microb Pathog. 1988 Aug;5(2):127–139. doi: 10.1016/0882-4010(88)90015-0. [DOI] [PubMed] [Google Scholar]

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