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. 1986 Dec;54(3):689–694. doi: 10.1128/iai.54.3.689-694.1986

Interaction of complement with serum-sensitive and serum-resistant strains of Pseudomonas aeruginosa.

N L Schiller, K A Joiner
PMCID: PMC260224  PMID: 3096887

Abstract

The interaction of complement with the following two strains of Pseudomonas aeruginosa was examined: 144M, a mucoid, serum-sensitive strain bearing short lipopolysaccharide O chains, and 144M-SR, a mucoid, serum-resistant strain bearing long lipopolysaccharide O chains isolated by repeated passage of 144M in increasing concentrations of pooled normal human serum (PNHS). While significant killing of 144M occurred in 5 to 40% PNHS, no killing of 144M-SR was observed. Both strains activated complement, especially 144M-SR which consumed 88.7, 96.4, and 100% of the available complement 3 (C3), C5, and C9, respectively, in 10% PNHS during a 60-min incubation at 37 degrees C. Although it activated more C3 than did 144M (54.9% consumption), 144M-SR bound only half as much C3 as 144M. Similarly, although 144M-SR activated more C9 than did 144M (50.0% consumption in 60 min), there was considerably less C9 attached to 144M-SR (2,990 molecules of C9 per bacterium) than to 144M (13,700 molecules per bacterium) after 60 min of incubation. Furthermore, only 162 molecules of the C9 bound to 144M-SR remained bound after treatment with 0.1% trypsin, while 5,692 molecules of the C9 bound to 144M remained bound under similar conditions. These results show that the serum resistance of 144M-SR does not represent a failure to activate complement efficiently, but instead reflects failure of the assembled terminal complement complex C5b-9 to insert stably into the outer membrane of this strain.

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Selected References

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  1. Agüero M. E., Aron L., DeLuca A. G., Timmis K. N., Cabello F. C. A plasmid-encoded outer membrane protein, TraT, enhances resistance of Escherichia coli to phagocytosis. Infect Immun. 1984 Dec;46(3):740–746. doi: 10.1128/iai.46.3.740-746.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Engels W., Endert J., Kamps M. A., van Boven C. P. Role of lipopolysaccharide in opsonization and phagocytosis of Pseudomonas aeruginosa. Infect Immun. 1985 Jul;49(1):182–189. doi: 10.1128/iai.49.1.182-189.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Engels W., Endert J., Van Boven C. P. A quantitative method for assessing the third complement factor (C3) attached to the surface of opsonized Pseudomonas aeruginosa: interrelationship between C3 fixation, phagocytosis and complement consumption. J Immunol Methods. 1985 Jul 16;81(1):43–53. doi: 10.1016/0022-1759(85)90120-6. [DOI] [PubMed] [Google Scholar]
  4. Fries L. F., Gaither T. A., Hammer C. H., Frank M. M. C3b covalently bound to IgG demonstrates a reduced rate of inactivation by factors H and I. J Exp Med. 1984 Dec 1;160(6):1640–1655. doi: 10.1084/jem.160.6.1640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Goldman J. N., Ruddy S., Austen K. F., Feingold D. S. The serum bactericidal reaction. 3. Antibody and complement requirements for killing a rough Escherichia coli. J Immunol. 1969 Jun;102(6):1379–1387. [PubMed] [Google Scholar]
  6. Goldman R. C., Joiner K., Leive L. Serum-resistant mutants of Escherichia coli O111 contain increased lipopolysaccharide, lack an O antigen-containing capsule, and cover more of their lipid A core with O antigen. J Bacteriol. 1984 Sep;159(3):877–882. doi: 10.1128/jb.159.3.877-882.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hammer C. H., Wirtz G. H., Renfer L., Gresham H. D., Tack B. F. Large scale isolation of functionally active components of the human complement system. J Biol Chem. 1981 Apr 25;256(8):3995–4006. [PubMed] [Google Scholar]
  8. Hancock R. E., Mutharia L. M., Chan L., Darveau R. P., Speert D. P., Pier G. B. Pseudomonas aeruginosa isolates from patients with cystic fibrosis: a class of serum-sensitive, nontypable strains deficient in lipopolysaccharide O side chains. Infect Immun. 1983 Oct;42(1):170–177. doi: 10.1128/iai.42.1.170-177.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hess K., Kern S., Schiller H. H. Blink reflex in trigeminal sensory neuropathy. Electromyogr Clin Neurophysiol. 1984 Mar-Apr;24(3):185–190. [PubMed] [Google Scholar]
  10. Holby N., Olling S. Pseudomonas aeruginosa infection in cystic fibrosis. Bactericidal effect of serum from normal individuals and patients with cystic fibrosis on P. aeruginosa strains from patients with cystic fibrosis or other diseases. Acta Pathol Microbiol Scand C. 1977 Apr;85(2):107–114. [PubMed] [Google Scholar]
  11. Inoue K., Yonemasu K., Takamizawa A., Amano T. [Studies on the immune bacteriolysis. XIV. Requirement of all nine components of complement for immune bacteriolysis]. Biken J. 1968 Sep;11(3):203–206. [PubMed] [Google Scholar]
  12. Joiner K. A., Grossman N., Schmetz M., Leive L. C3 binds preferentially to long-chain lipopolysaccharide during alternative pathway activation by Salmonella montevideo. J Immunol. 1986 Jan;136(2):710–715. [PubMed] [Google Scholar]
  13. Joiner K. A., Hammer C. H., Brown E. J., Frank M. M. Studies on the mechanism of bacterial resistance to complement-mediated killing. II. C8 and C9 release C5b67 from the surface of Salmonella minnesota S218 because the terminal complex does not insert into the bacterial outer membrane. J Exp Med. 1982 Mar 1;155(3):809–819. doi: 10.1084/jem.155.3.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Joiner K. A., Schmetz M. A., Goldman R. C., Leive L., Frank M. M. Mechanism of bacterial resistance to complement-mediated killing: inserted C5b-9 correlates with killing for Escherichia coli O111B4 varying in O-antigen capsule and O-polysaccharide coverage of lipid A core oligosaccharide. Infect Immun. 1984 Jul;45(1):113–117. doi: 10.1128/iai.45.1.113-117.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Joiner K. A. Studies on the mechanism of bacterial resistance to complement-mediated killing and on the mechanism of action of bactericidal antibody. Curr Top Microbiol Immunol. 1985;121:99–133. doi: 10.1007/978-3-642-45604-6_6. [DOI] [PubMed] [Google Scholar]
  16. Leive L. Release of lipopolysaccharide by EDTA treatment of E. coli. Biochem Biophys Res Commun. 1965 Nov 22;21(4):290–296. doi: 10.1016/0006-291x(65)90191-9. [DOI] [PubMed] [Google Scholar]
  17. Meshulam T., Verbrugh H., Verhoef J. Serum-induced lysis of Pseudomonas aeruginosa. Eur J Clin Microbiol. 1982 Feb;1(1):1–6. doi: 10.1007/BF02014132. [DOI] [PubMed] [Google Scholar]
  18. Moll A., Manning P. A., Timmis K. N. Plasmid-determined resistance to serum bactericidal activity: a major outer membrane protein, the traT gene product, is responsible for plasmid-specified serum resistance in Escherichia coli. Infect Immun. 1980 May;28(2):359–367. doi: 10.1128/iai.28.2.359-367.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Muschel L. H., Ahl L. A., Fisher M. W. Sensitivity of Pseudomonas aeruginosa to normal serum and to polymyxin. J Bacteriol. 1969 May;98(2):453–457. doi: 10.1128/jb.98.2.453-457.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Muschel L. H., Larsen L. J. The sensitivity of smooth and rough gram-negative bacteria to the immune bactericidal reaction. Proc Soc Exp Biol Med. 1970 Jan;133(1):345–348. doi: 10.3181/00379727-133-34472. [DOI] [PubMed] [Google Scholar]
  21. Nelson B. W., Roantree R. J. Analyses of lipopolysaccharides extracted from penicillin-resistant, serum-sensitive salmonella mutants. J Gen Microbiol. 1967 Aug;48(2):179–188. doi: 10.1099/00221287-48-2-179. [DOI] [PubMed] [Google Scholar]
  22. Reyes M. P., El-Khatib M. R., Brown W. J., Smith F., Lerner A. M. Synergy between carbenicillin and an aminoglycoside (gentamicin or tobramycin) against Pseudomonas aeruginosa isolated from patients with endocarditis and sensitivity of isolates to normal human serum. J Infect Dis. 1979 Aug;140(2):192–202. doi: 10.1093/infdis/140.2.192. [DOI] [PubMed] [Google Scholar]
  23. Rowley D. Sensitivity of rough gram-negative bacteria to the bactericidal action of serum. J Bacteriol. 1968 May;95(5):1647–1650. doi: 10.1128/jb.95.5.1647-1650.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schiller N. L., Alazard M. J., Borowski R. S. Serum sensitivity of a Pseudomonas aeruginosa mucoid strain. Infect Immun. 1984 Sep;45(3):748–755. doi: 10.1128/iai.45.3.748-755.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schiller N. L., Hatch R. A. The serum sensitivity, colonial morphology, serogroup specificity, and outer membrane protein of Pseudomonas aeruginosa strains isolated from several clinical sites. Diagn Microbiol Infect Dis. 1983 Jun;1(2):145–157. doi: 10.1016/0732-8893(83)90044-5. [DOI] [PubMed] [Google Scholar]
  26. Schreiber R. D., Morrison D. C., Podack E. R., Müller-Eberhard H. J. Bactericidal activity of the alternative complement pathway generated from 11 isolated plasma proteins. J Exp Med. 1979 Apr 1;149(4):870–882. doi: 10.1084/jem.149.4.870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schultz D. R., Miller K. D. Elastase of Pseudomonas aeruginosa: inactivation of complement components and complement-derived chemotactic and phagocytic factors. Infect Immun. 1974 Jul;10(1):128–135. doi: 10.1128/iai.10.1.128-135.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Taylor P. W., Kroll H. P. Interaction of human complement proteins with serum-sensitive and serum-resistant strains of Escherichia coli. Mol Immunol. 1984 Jul;21(7):609–620. doi: 10.1016/0161-5890(84)90046-4. [DOI] [PubMed] [Google Scholar]
  29. Taylor P. W., Robinson M. K. Determinants that increase the serum resistance of Escherichia coli. Infect Immun. 1980 Jul;29(1):278–280. doi: 10.1128/iai.29.1.278-280.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Thomassen M. J., Demko C. A. Serum bactericidal effect on Pseudomonas aeruginosa isolates from cystic fibrosis patients. Infect Immun. 1981 Aug;33(2):512–518. doi: 10.1128/iai.33.2.512-518.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Young L. S., Armstrong D. Human immunity to Pseudomonas aeruginosa. I. In-vitro interaction of bacteria, polymorphonuclear leukocytes, and serum factors. J Infect Dis. 1972 Sep;126(3):257–276. doi: 10.1093/infdis/126.3.257. [DOI] [PubMed] [Google Scholar]

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