Immunoelectrophoretic analysis, toxicity, and kinetics of in vitro production of the protective antigen and lethal factor components of Bacillus anthracis toxin - PubMed (original) (raw)

Immunoelectrophoretic analysis, toxicity, and kinetics of in vitro production of the protective antigen and lethal factor components of Bacillus anthracis toxin

J W Ezzell et al. Infect Immun. 1984 Sep.

Abstract

The kinetics of Bacillus anthracis toxin production in culture and its lethal activity in rats, mice, and guinea pigs were investigated. Lethal toxin activity was produced in vitro throughout exponential growth at essentially identical rates in both encapsulated virulent and nonencapsulated avirulent strains. The two toxin proteins which produce lethality when in combination, lethal factor (LF) and protective antigen (PA), could be quantitated directly from culture fluids by rocket immunoelectrophoresis. Using purified preparations of these proteins, we determined that a combination of 8 micrograms of LF and 40 micrograms of PA was required for a maximal rate of killing (39 to 40 min) in Fischer 344 rats (250 to 300 g). Conversely, a minimum of 0.6 microgram of LF and 3 micrograms of PA was required for lethality. The 50% lethal dose for Hartley guinea pigs was 50 micrograms of LF and 250 micrograms of PA, and for Swiss mice it was 2.5 micrograms of LF and 12.5 micrograms of PA. Analyses classically reserved for enzyme kinetic studies were used to study the kinetics of lethal activity in the rat model after intravenous injection of LF-PA mixtures. The amounts of LF and PA which were required to give half the rate of killing (i.e., double the minimum time to death) were 1.2 and 5.8 micrograms, respectively. A theoretical minimum time to death was determined to be 38 min. A third anthrax toxin component, edema factor, was shown to inhibit lethal toxin activity. Edema factor could not be quantitated by rocket immunoelectrophoresis because the protein did not form distinct precipitin bands with available antisera.

PubMed Disclaimer

Similar articles

• Contribution of individual toxin components to virulence of Bacillus anthracis.
  Pezard C, Berche P, Mock M. Pezard C, et al. Infect Immun. 1991 Oct;59(10):3472-7. doi: 10.1128/iai.59.10.3472-3477.1991. Infect Immun. 1991. PMID: 1910002 Free PMC article.
• Role of toxin functional domains in anthrax pathogenesis.
  Brossier F, Weber-Levy M, Mock M, Sirard JC. Brossier F, et al. Infect Immun. 2000 Apr;68(4):1781-6. doi: 10.1128/IAI.68.4.1781-1786.2000. Infect Immun. 2000. PMID: 10722564 Free PMC article.
• A new triple chimeric protein as a high immunogenic antigen against anthrax toxins: theoretical and experimental analyses.
  Abdous M, Hasannia S, Salmanian AH, Shahryar Arab S, Shali A, Alizadeh GA, Hajizadeh A, Khafri A, Mohseni A. Abdous M, et al. Immunopharmacol Immunotoxicol. 2019 Feb;41(1):25-31. doi: 10.1080/08923973.2018.1510419. Epub 2019 Jan 9. Immunopharmacol Immunotoxicol. 2019. PMID: 30621469
• Anthrax toxin.
  Bhatnagar R, Batra S. Bhatnagar R, et al. Crit Rev Microbiol. 2001;27(3):167-200. doi: 10.1080/20014091096738. Crit Rev Microbiol. 2001. PMID: 11596878 Review.
• Anthrax toxin: a tripartite lethal combination.
  Ascenzi P, Visca P, Ippolito G, Spallarossa A, Bolognesi M, Montecucco C. Ascenzi P, et al. FEBS Lett. 2002 Nov 20;531(3):384-8. doi: 10.1016/s0014-5793(02)03609-8. FEBS Lett. 2002. PMID: 12435580 Review.

Cited by

• Inhibitory Effects of a Reengineered Anthrax Toxin on Canine and Human Osteosarcoma Cells.
  Mackowiak da Fonseca J, Mackowiak da Fonseca II, Nagamine MK, Massoco CO, Nishiya AT, Ward JM, Liu S, Leppla SH, Bugge TH, Dagli MLZ. Mackowiak da Fonseca J, et al. Toxins (Basel). 2020 Sep 24;12(10):614. doi: 10.3390/toxins12100614. Toxins (Basel). 2020. PMID: 32987941 Free PMC article.
• Anthrax toxins induce shock in rats by depressed cardiac ventricular function.
  Watson LE, Kuo SR, Katki K, Dang T, Park SK, Dostal DE, Tang WJ, Leppla SH, Frankel AE. Watson LE, et al. PLoS One. 2007 May 23;2(5):e466. doi: 10.1371/journal.pone.0000466. PLoS One. 2007. PMID: 17520025 Free PMC article.
• Role of N-terminal amino acids in the potency of anthrax lethal factor.
  Gupta PK, Moayeri M, Crown D, Fattah RJ, Leppla SH. Gupta PK, et al. PLoS One. 2008 Sep 3;3(9):e3130. doi: 10.1371/journal.pone.0003130. PLoS One. 2008. PMID: 18769623 Free PMC article.
• Recent advances in the development of an improved, human anthrax vaccine.
  Ivins BE, Welkos SL. Ivins BE, et al. Eur J Epidemiol. 1988 Mar;4(1):12-9. doi: 10.1007/BF00152686. Eur J Epidemiol. 1988. PMID: 3128450 Review.
• Anthrax lethal toxin induces ketotifen-sensitive intradermal vascular leakage in certain inbred mice.
  Gozes Y, Moayeri M, Wiggins JF, Leppla SH. Gozes Y, et al. Infect Immun. 2006 Feb;74(2):1266-72. doi: 10.1128/IAI.74.2.1266-1272.2006. Infect Immun. 2006. PMID: 16428776 Free PMC article.

References

1. 1. Microbiol Rev. 1982 Mar;46(1):86-94 - PubMed
2. 1. Appl Environ Microbiol. 1981 Jun;41(6):1479-81 - PubMed
3. 1. Infect Immun. 1983 Jan;39(1):371-6 - PubMed
4. 1. Infect Immun. 1983 Jan;39(1):483-6 - PubMed
5. 1. J Exp Med. 1954 Feb;99(2):167-82 - PubMed

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Atypon
  • Europe PubMed Central
  • PubMed Central

• Other Literature Sources

  • The Lens - Patent Citations Database