Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1988 Nov;56(11):2788–2793. doi: 10.1128/iai.56.11.2788-2793.1988

Alginase treatment of mucoid Pseudomonas aeruginosa enhances phagocytosis by human monocyte-derived macrophages.

F Eftekhar 1, D P Speert 1
PMCID: PMC259651  PMID: 3139564

Abstract

Mucoid Pseudomonas aeruginosa colonizes and infects the respiratory tract of most older patients with cystic fibrosis. These bacteria resist both opsonin-dependent and -independent phagocytosis by human polymorphonuclear leukocytes and monocyte-derived macrophages. Resistance to phagocytosis is thought to be mediated in part by the mucoid exopolysaccharide associated with the bacterial surface. The purpose of this study was to determine whether degradation of the mucoid exopolysaccharide by alginase enhances bacterial susceptibility to nonopsonic phagocytosis by macrophages. Eight phagocytosis-resistant mucoid P. aeruginosa isolates from patients with cystic fibrosis were studied. The bacteria were treated with a preparation of alginase from Bacillus circulans, and phagocytosis by macrophages was measured by a visual inspection assay. Alginase degradation of mucoid exopolysaccharide was measured by the periodate-thiobarbituric acid assay and by indirect immunofluorescence with a mouse monoclonal antibody to the mucoid exopolysaccharide. Alginase degraded the mucoid exopolysaccharide of all eight mucoid strains tested. Phagocytosis was enhanced in five of the eight strains. Alginase-enhanced phagocytosis was magnesium dependent and heat labile. Alginase may be a useful tool for studying the biological properties of P. aeruginosa mucoid exopolysaccharide.

Full text

PDF
2788

Images in this article

2790Image
on p.2790

Selected References

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

  1. Baltimore R. S., Mitchell M. Immunologic investigations of mucoid strains of Pseudomonas aeruginosa: comparison of susceptibility to opsonic antibody in mucoid and nonmucoid strains. J Infect Dis. 1980 Feb;141(2):238–247. doi: 10.1093/infdis/141.2.238. [DOI] [PubMed] [Google Scholar]
  2. Boyd J., Turvey J. R. Isolation of poly-alpha-L-guluronate lyase from Klebsiella aerogenes. Carbohydr Res. 1977 Aug;57:163–171. doi: 10.1016/s0008-6215(00)81928-x. [DOI] [PubMed] [Google Scholar]
  3. Cabral D. A., Loh B. A., Speert D. P. Mucoid Pseudomonas aeruginosa resists nonopsonic phagocytosis by human neutrophils and macrophages. Pediatr Res. 1987 Oct;22(4):429–431. doi: 10.1203/00006450-198710000-00013. [DOI] [PubMed] [Google Scholar]
  4. Davidson I. W., Lawson C. J., Sutherland I. W. An alginate lysate from Azotobacter vinelandii phage. J Gen Microbiol. 1977 Jan;98(1):223–229. doi: 10.1099/00221287-98-1-223. [DOI] [PubMed] [Google Scholar]
  5. Davidson I. W., Sutherland I. W., Lawson C. J. Purification and properties of an alginate lyase from a marine bacterium. Biochem J. 1976 Dec 1;159(3):707–713. doi: 10.1042/bj1590707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Doig P., Smith N. R., Todd T., Irvin R. T. Characterization of the binding of Pseudomonas aeruginosa alginate to human epithelial cells. Infect Immun. 1987 Jun;55(6):1517–1522. doi: 10.1128/iai.55.6.1517-1522.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Doubet R. S., Quatrano R. S. Isolation of marine bacteria capable of producing specific lyases for alginate degradation. Appl Environ Microbiol. 1982 Sep;44(3):754–756. doi: 10.1128/aem.44.3.754-756.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dunne W. M., Jr, Buckmire F. L. Partial purification and characterization of a polymannuronic acid depolymerase produced by a mucoid strain of Pseudomonas aeruginosa isolated from a patient with cystic fibrosis. Appl Environ Microbiol. 1985 Sep;50(3):562–567. doi: 10.1128/aem.50.3.562-567.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evans L. R., Linker A. Production and characterization of the slime polysaccharide of Pseudomonas aeruginosa. J Bacteriol. 1973 Nov;116(2):915–924. doi: 10.1128/jb.116.2.915-924.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hansen J. B., Doubet R. S., Ram J. Alginase enzyme production by Bacillus circulans. Appl Environ Microbiol. 1984 Apr;47(4):704–709. doi: 10.1128/aem.47.4.704-709.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hansen J. B., Nakamura L. K. Distribution of Alginate Lyase Activity among Strains of Bacillus circulans. Appl Environ Microbiol. 1985 Apr;49(4):1019–1021. doi: 10.1128/aem.49.4.1019-1021.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Haug A., Larsen B. Biosynthesis of alginate. II. Polymannuronic acid C-5-epimerase from Azotobacter vinelandii (Lipman). Carbohydr Res. 1971 Apr;17(2):297–308. doi: 10.1016/s0008-6215(00)82537-9. [DOI] [PubMed] [Google Scholar]
  13. Kashiwabara Y., Suzuki H., Nisizawa K. Alginate lyases of pseudomonads. J Biochem. 1969 Oct;66(4):503–512. doi: 10.1093/oxfordjournals.jbchem.a129175. [DOI] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  16. Linker A., Evans L. R. Isolation and characterization of an alginase from mucoid strains of Pseudomonas aeruginosa. J Bacteriol. 1984 Sep;159(3):958–964. doi: 10.1128/jb.159.3.958-964.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Linker A., Jones R. S. A new polysaccharide resembling alginic acid isolated from pseudomonads. J Biol Chem. 1966 Aug 25;241(16):3845–3851. [PubMed] [Google Scholar]
  18. Mutharia L. M., Hancock R. E. Surface localization of Pseudomonas aeruginosa outer membrane porin protein F by using monoclonal antibodies. Infect Immun. 1983 Dec;42(3):1027–1033. doi: 10.1128/iai.42.3.1027-1033.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. PREISS J., ASHWELL G. Alginic acid metabolism in bacteria. I. Enzymatic formation of unsaturated oligosac-charides and 4-deoxy-L-erythro-5-hexoseulose uronic acid. J Biol Chem. 1962 Feb;237:309–316. [PubMed] [Google Scholar]
  20. Pennington J. E., Wolff S. M., Puziss M. Summary of a workshop on infections in patients with cystic fibrosis. J Infect Dis. 1979 Aug;140(2):252–256. doi: 10.1093/infdis/140.2.252. [DOI] [PubMed] [Google Scholar]
  21. Ramphal R., Pier G. B. Role of Pseudomonas aeruginosa mucoid exopolysaccharide in adherence to tracheal cells. Infect Immun. 1985 Jan;47(1):1–4. doi: 10.1128/iai.47.1.1-4.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sherbrock-Cox V., Russell N. J., Gacesa P. The purification and chemical characterisation of the alginate present in extracellular material produced by mucoid strains of Pseudomonas aeruginosa. Carbohydr Res. 1984 Dec 15;135(1):147–154. doi: 10.1016/0008-6215(84)85012-0. [DOI] [PubMed] [Google Scholar]
  23. Skjåk-Braek G., Grasdalen H., Larsen B. Monomer sequence and acetylation pattern in some bacterial alginates. Carbohydr Res. 1986 Oct 15;154:239–250. doi: 10.1016/s0008-6215(00)90036-3. [DOI] [PubMed] [Google Scholar]
  24. Speert D. P., Lawton D., Mutharia L. M. Antibody to Pseudomonas aeruginosa mucoid exopolysaccharide and to sodium alginate in cystic fibrosis serum. Pediatr Res. 1984 May;18(5):431–433. doi: 10.1203/00006450-198405000-00008. [DOI] [PubMed] [Google Scholar]
  25. Speert D. P., Loh B. A., Cabral D. A., Salit I. E. Nonopsonic phagocytosis of nonmucoid Pseudomonas aeruginosa by human neutrophils and monocyte-derived macrophages is correlated with bacterial piliation and hydrophobicity. Infect Immun. 1986 Jul;53(1):207–212. doi: 10.1128/iai.53.1.207-212.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Speert D. P., Silverstein S. C. Phagocytosis of unopsonized zymosan by human monocyte-derived macrophages: maturation and inhibition by mannan. J Leukoc Biol. 1985 Nov;38(5):655–658. doi: 10.1002/jlb.38.5.655. [DOI] [PubMed] [Google Scholar]
  27. Stevens R. A., Levin R. E. Purification and characteristics of an alginase from Alginovibrio aquatilis. Appl Environ Microbiol. 1977 May;33(5):1156–1161. doi: 10.1128/aem.33.5.1156-1161.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. WEISSBACH A., HURWITZ J. The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. I. Identification. J Biol Chem. 1959 Apr;234(4):705–709. [PubMed] [Google Scholar]
  29. von Riesen V. L. Digestion of algin by Pseudomonas maltophilia and Pseudomonas putida. Appl Environ Microbiol. 1980 Jan;39(1):92–96. doi: 10.1128/aem.39.1.92-96.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES