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. 1988 Jul;56(7):1807–1813. doi: 10.1128/iai.56.7.1807-1813.1988

Production and characterization of monoclonal antibodies to the protective antigen component of Bacillus anthracis toxin.

S F Little 1, S H Leppla 1, E Cora 1
PMCID: PMC259481  PMID: 3384478

Abstract

Thirty-six monoclonal antibodies to the protective antigen protein of Bacillus anthracis exotoxin have been characterized for affinity, antibody subtype, competitive binding to antigenic regions, and ability to neutralize lethal and edema toxin activities. At least 23 antigenic regions were detected on protective antigen by a blocking, enzyme-linked immunosorbent assay. Two clones, 3B6 and 14B7, competed for a single antigenic region and neutralized the activity of both the lethal toxin in vivo (Fisher 344 rat) and the edema toxin in vitro (CHO cells). These two antibodies blocked the binding of 125I-labeled protective antigen to FRL-103 cells. Our results support the proposal that binding of protective antigen to cell receptors is required for expression of toxicity.

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Selected References

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  1. Adám E., Nász I., Lengyel A., Erdei J., Fachet J. Determination of different antigenic sites on the adenovirus hexon using monoclonal antibodies. Arch Virol. 1987;93(3-4):261–271. doi: 10.1007/BF01310979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Carbone F. R., Paterson Y. Monoclonal antibodies to horse cytochrome c expressing four distinct idiotypes distribute among two sites on the native protein. J Immunol. 1985 Oct;135(4):2609–2616. [PubMed] [Google Scholar]
  4. Cianfriglia M., Armellini D., Massone A., Mariani M. Simple immunization protocol for high frequency production of soluble antigen-specific hybridomas. Hybridoma. 1983;2(4):451–457. doi: 10.1089/hyb.1983.2.451. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Friedlander A. M. Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process. J Biol Chem. 1986 Jun 5;261(16):7123–7126. [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Huber-Lukac M., Jaquet F., Luethy P., Huetter R., Braun D. G. Characterization of monoclonal antibodies to a crystal protein of Bacillus thuringiensis subsp. kurstaki. Infect Immun. 1986 Oct;54(1):228–232. doi: 10.1128/iai.54.1.228-232.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Huber-Lukac M., Lüthy P., Braun D. G. Specificities of monoclonal antibodies against the activated delta-endotoxin of Bacillus thuringiensis var. thuringiensis. Infect Immun. 1983 May;40(2):608–612. doi: 10.1128/iai.40.2.608-612.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. MOLNAR D. M., ALTENBERN R. A. ALTERATIONS IN THE BIOLOGICAL ACTIVITY OF PROTECTIVE ANTIGEN OF BACILLUS ANTHRACIS TOXIN. Proc Soc Exp Biol Med. 1963 Nov;114:294–297. [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. Ray K., Vockley J., Harris H. Epitopes of human intestinal alkaline phosphatases, defined by monoclonal antibodies. FEBS Lett. 1984 Sep 3;174(2):294–299. doi: 10.1016/0014-5793(84)81176-x. [DOI] [PubMed] [Google Scholar]
  17. STANLEY J. L., SARGEANT K., SMITH H. Purification of factors I and II of the anthrax toxin produced in vivo. J Gen Microbiol. 1960 Feb;22:206–218. doi: 10.1099/00221287-22-1-206. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Sheppard A. J., Cussell D., Hughes M. Production and characterization of monoclonal antibodies to tetanus toxin. Infect Immun. 1984 Feb;43(2):710–714. doi: 10.1128/iai.43.2.710-714.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Smith-Gill S. J., Lavoie T. B., Mainhart C. R. Antigenic regions defined by monoclonal antibodies correspond to structural domains of avian lysozyme. J Immunol. 1984 Jul;133(1):384–393. [PubMed] [Google Scholar]
  21. Stähli C., Miggiano V., Stocker J., Staehelin T., Häring P., Takács B. Distinction of epitopes by monoclonal antibodies. Methods Enzymol. 1983;92:242–253. doi: 10.1016/0076-6879(83)92023-2. [DOI] [PubMed] [Google Scholar]
  22. THORNE C. B., MOLNAR D. M., STRANGE R. E. Production of toxin in vitro by Bacillus anthracis and its spearation into two components. J Bacteriol. 1960 Mar;79:450–455. doi: 10.1128/jb.79.3.450-455.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tzartos S. J., Rand D. E., Einarson B. L., Lindstrom J. M. Mapping of surface structures of electrophorus acetylcholine receptor using monoclonal antibodies. J Biol Chem. 1981 Aug 25;256(16):8635–8645. [PubMed] [Google Scholar]
  24. Uchida I., Sekizaki T., Hashimoto K., Terakado N. Association of the encapsulation of Bacillus anthracis with a 60 megadalton plasmid. J Gen Microbiol. 1985 Feb;131(2):363–367. doi: 10.1099/00221287-131-2-363. [DOI] [PubMed] [Google Scholar]
  25. Van Heyningen V., Brock D. J., Van Heyningen S. A simple method for ranking the affinities of monoclonal antibodies. J Immunol Methods. 1983 Aug 26;62(2):147–153. doi: 10.1016/0022-1759(83)90000-5. [DOI] [PubMed] [Google Scholar]
  26. Volk W. A., Bizzini B., Snyder R. M., Bernhard E., Wagner R. R. Neutralization of tetanus toxin by distinct monoclonal antibodies binding to multiple epitopes on the toxin molecule. Infect Immun. 1984 Sep;45(3):604–609. doi: 10.1128/iai.45.3.604-609.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wnek A. P., Strouse R. J., McClane B. A. Production and characterization of monoclonal antibodies against Clostridium perfringens type A enterotoxin. Infect Immun. 1985 Nov;50(2):442–448. doi: 10.1128/iai.50.2.442-448.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wright G. G., Mandell G. L. Anthrax toxin blocks priming of neutrophils by lipopolysaccharide and by muramyl dipeptide. J Exp Med. 1986 Nov 1;164(5):1700–1709. doi: 10.1084/jem.164.5.1700. [DOI] [PMC free article] [PubMed] [Google Scholar]

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