Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1984 Feb;43(2):700–705. doi: 10.1128/iai.43.2.700-705.1984

Extraction and isolation of a leukotoxin from Actinobacillus actinomycetemcomitans with polymyxin B.

C C Tsai, B J Shenker, J M DiRienzo, D Malamud, N S Taichman
PMCID: PMC264356  PMID: 6319288

Abstract

A leukotoxin from Actinobacillus actinomycetemcomitans was isolated by a procedure that includes polymyxin B extraction, ion-exchange chromatography, and gel filtration chromatography. The procedure resulted in the recovery of 48% of the toxin with a 99-fold increase in specific activity. The isolated toxin has a molecular mass of 180,000 daltons by gel filtration and 115,000 daltons by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It retains all the major biological characteristics previously documented for crude leukotoxin preparations, including susceptibility to heat and proteolytic enzymes and neutralization by sera from patients with juvenile periodontitis. The isolated leukotoxin destroys human but not rat or guinea pig polymorphonuclear leukocytes and has no apparent effect on human erythrocytes. The availability of the A. actinomycetemcomitans leukotoxin should facilitate studies on its chemistry and mode of action as well as its role in the pathogenesis of human periodontal disease.

Full text

PDF
700

Images in this article

703Image
on p.703

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baehni P. C., Tsai C. C., McArthur W. P., Hammond B. F., Shenker B. J., Taichman N. S. Leukotoxic activity in different strains of the bacterium Actinobacillus actinomycetemcomitans isolated from juvenile periodontitis in man. Arch Oral Biol. 1981;26(8):671–676. doi: 10.1016/0003-9969(81)90164-3. [DOI] [PubMed] [Google Scholar]
  2. Baehni P., Tsai C. C., McArthur W. P., Hammond B. F., Taichman N. S. Interaction of inflammatory cells and oral microorganisms. VIII. Detection of leukotoxic activity of a plaque-derived gram-negative microorganism. Infect Immun. 1979 Apr;24(1):233–243. doi: 10.1128/iai.24.1.233-243.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Di Rienzo J. M., Spieler E. L. Identification and characterization of the major cell envelope proteins of oral strains of Actinobacillus actinomycetemcomitans. Infect Immun. 1983 Jan;39(1):253–261. doi: 10.1128/iai.39.1.253-261.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Donohue-Rolfe A., Keusch G. T. Shigella dysenteriae 1 cytotoxin: periplasmic protein releasable by polymyxin B and osmotic shock. Infect Immun. 1983 Jan;39(1):270–274. doi: 10.1128/iai.39.1.270-274.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ebersole J. L., Taubman M. A., Smith D. J., Genco R. J., Frey D. E. Human immune responses to oral micro-organisms. I. Association of localized juvenile periodontitis (LJP) with serum antibody responses to Actinobacillus actinomycetemcomitans. Clin Exp Immunol. 1982 Jan;47(1):43–52. [PMC free article] [PubMed] [Google Scholar]
  6. Evans D. J., Evans D. G., Gorbach S. L. Polymyxin B-Induced Release of Low-Molecular-Weight, Heat-Labile Enterotoxin from Escherichia coli. Infect Immun. 1974 Nov;10(5):1010–1017. doi: 10.1128/iai.10.5.1010-1017.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Griffin D. E., Gemski P. Release of Shiga toxin from Shigella dysenteriae 1 by polymyxin B. Infect Immun. 1983 Apr;40(1):425–428. doi: 10.1128/iai.40.1.425-428.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kiley P., Holt S. C. Characterization of the lipopolysaccharide from Actinobacillus actinomycetemcomitans Y4 and N27. Infect Immun. 1980 Dec;30(3):862–873. doi: 10.1128/iai.30.3.862-873.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Listgarten M. A., Lai C. H., Evian C. I. Comparative antibody titers to Actinobacillus actinomycetemcomitans in juvenile periodontitis, chronic periodontitis and periodontally healthy subjects. J Clin Periodontol. 1981 Jun;8(3):155–164. doi: 10.1111/j.1600-051x.1981.tb02027.x. [DOI] [PubMed] [Google Scholar]
  10. Lockwood D. E., Kreger A. S., Richardson S. H. Detection of toxins produced by vibrio fluvialis. Infect Immun. 1982 Feb;35(2):702–708. doi: 10.1128/iai.35.2.702-708.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Moore W. E., Holdeman L. V., Smibert R. M., Hash D. E., Burmeister J. A., Ranney R. R. Bacteriology of severe periodontitis in young adult humans. Infect Immun. 1982 Dec;38(3):1137–1148. doi: 10.1128/iai.38.3.1137-1148.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Newman M. G., Socransky S. S., Savitt E. D., Propas D. A., Crawford A. Studies of the microbiology of periodontosis. J Periodontol. 1976 Jul;47(7):373–379. doi: 10.1902/jop.1976.47.7.373. [DOI] [PubMed] [Google Scholar]
  13. Nowotny A., Behling U. H., Hammond B., Lai C. H., Listgarten M., Pham P. H., Sanavi F. Release of toxic microvesicles by Actinobacillus actinomycetemcomitans. Infect Immun. 1982 Jul;37(1):151–154. doi: 10.1128/iai.37.1.151-154.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ranney R. R., Yanni N. R., Burmeister J. A., Tew J. G. Relationship between attachment loss and precipitating serum antibody to Actinobacillus actinomycetemcomitans in adolescents and young adults having severe periodontal destruction. J Periodontol. 1982 Jan;53(1):1–7. doi: 10.1902/jop.1982.53.1.1. [DOI] [PubMed] [Google Scholar]
  15. Robertson P. B., Lantz M., Marucha P. T., Kornman K. S., Trummel C. L., Holt S. C. Collagenolytic activity associated with Bacteroides species and Actinobacillus actinomycetemcomitans. J Periodontal Res. 1982 May;17(3):275–283. doi: 10.1111/j.1600-0765.1982.tb01154.x. [DOI] [PubMed] [Google Scholar]
  16. Shenker B. J., Kushner M. E., Tsai C. C. Inhibition of fibroblast proliferation by Actinobacillus actinomycetemcomitans. Infect Immun. 1982 Dec;38(3):986–992. doi: 10.1128/iai.38.3.986-992.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Shenker B. J., McArthur W. P., Tsai C. C. Immune suppression induced by Actinobacillus actinomycetemcomitans. I. Effects on human peripheral blood lymphocyte responses to mitogens and antigens. J Immunol. 1982 Jan;128(1):148–154. [PubMed] [Google Scholar]
  18. Slots J., Reynolds H. S., Genco R. J. Actinobacillus actinomycetemcomitans in human periodontal disease: a cross-sectional microbiological investigation. Infect Immun. 1980 Sep;29(3):1013–1020. doi: 10.1128/iai.29.3.1013-1020.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Slots J., Zambon J. J., Rosling B. G., Reynolds H. S., Christersson L. A., Genco R. J. Actinobacillus actinomycetemcomitans in human periodontal disease. Association, serology, leukotoxicity, and treatment. J Periodontal Res. 1982 Sep;17(5):447–448. doi: 10.1111/j.1600-0765.1982.tb02022.x. [DOI] [PubMed] [Google Scholar]
  20. Taichman N. S., Dean R. T., Sanderson C. J. Biochemical and morphological characterization of the killing of human monocytes by a leukotoxin derived from Actinobacillus actinomycetemcomitans. Infect Immun. 1980 Apr;28(1):258–268. doi: 10.1128/iai.28.1.258-268.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Taichman N. S., Wilton J. M. Leukotoxicity of an extract from Actinobacillus actinomycetemcomitans for human gingival polymorphonuclear leukocytes. Inflammation. 1981 Mar;5(1):1–12. doi: 10.1007/BF00910774. [DOI] [PubMed] [Google Scholar]
  22. Tanner A. C., Haffer C., Bratthall G. T., Visconti R. A., Socransky S. S. A study of the bacteria associated with advancing periodontitis in man. J Clin Periodontol. 1979 Oct;6(5):278–307. doi: 10.1111/j.1600-051x.1979.tb01931.x. [DOI] [PubMed] [Google Scholar]
  23. Tsai C. C., McArthur W. P., Baehni P. C., Evian C., Genco R. J., Taichman N. S. Serum neutralizing activity against Actinobacillus actinomycetemcomitans leukotoxin in juvenile periodontitis. J Clin Periodontol. 1981 Aug;8(4):338–348. doi: 10.1111/j.1600-051x.1981.tb02043.x. [DOI] [PubMed] [Google Scholar]
  24. Tsai C. C., McArthur W. P., Baehni P. C., Hammond B. F., Taichman N. S. Extraction and partial characterization of a leukotoxin from a plaque-derived Gram-negative microorganism. Infect Immun. 1979 Jul;25(1):427–439. doi: 10.1128/iai.25.1.427-439.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Zambon J. J., DeLuca C., Slots J., Genco R. J. Studies of leukotoxin from Actinobacillus actinomycetemcomitans using the promyelocytic HL-60 cell line. Infect Immun. 1983 Apr;40(1):205–212. doi: 10.1128/iai.40.1.205-212.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zambon J. J., Slots J., Genco R. J. Serology of oral Actinobacillus actinomycetemcomitans and serotype distribution in human periodontal disease. Infect Immun. 1983 Jul;41(1):19–27. doi: 10.1128/iai.41.1.19-27.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES