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. 1989 Dec;57(12):3841–3845. doi: 10.1128/iai.57.12.3841-3845.1989

Pseudomonas aeruginosa pili as ligands for nonopsonic phagocytosis by fibronectin-stimulated macrophages.

N M Kelly 1, J L Kluftinger 1, B L Pasloske 1, W Paranchych 1, R E Hancock 1
PMCID: PMC259914  PMID: 2572562

Abstract

Fibronectin is capable of activating macrophages for enhanced nonopsonic phagocytosis of Pseudomonas aeruginosa grown in vivo in rats or mice or in vitro on nutrient agar plates. In this study it was determined that while fibronectin was able to significantly increase phagocytosis of organisms grown in static broth, uptake of agitated bacteria could not be promoted. Agitated P. aeruginosa cultures were proven to lack surface pili expression, as assessed by electron microscopic studies. A pilus-deficient pilA::Tn501 mutant of P. aeruginosa PAO was constructed by gene replacement techniques. Phagocytosis of this mutant could not be enhanced by fibronectin regardless of growth conditions. Furthermore, 60 micrograms of exogenously added Pseudomonas pili per ml was capable of abrogating the enhanced phagocytosis of the wild-type strain observed with fibronectin-stimulated macrophages. It is concluded that Pseudomonas pili were the bacterial ligands required for attachment to fibronectin-stimulated macrophages in the initial stages of nonopsonic phagocytosis.

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Selected References

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  1. Bagdasarian M., Lurz R., Rückert B., Franklin F. C., Bagdasarian M. M., Frey J., Timmis K. N. Specific-purpose plasmid cloning vectors. II. Broad host range, high copy number, RSF1010-derived vectors, and a host-vector system for gene cloning in Pseudomonas. Gene. 1981 Dec;16(1-3):237–247. doi: 10.1016/0378-1119(81)90080-9. [DOI] [PubMed] [Google Scholar]
  2. Battershill J. L., Speert D. P., Hancock R. E. Use of monoclonal antibodies to protein F of Pseudomonas aeruginosa as opsonins for phagocytosis by macrophages. Infect Immun. 1987 Oct;55(10):2531–2533. doi: 10.1128/iai.55.10.2531-2533.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blumenstock E., Jann K. Adhesion of piliated Escherichia coli strains to phagocytes: differences between bacteria with mannose-sensitive pili and those with mannose-resistant pili. Infect Immun. 1982 Jan;35(1):264–269. doi: 10.1128/iai.35.1.264-269.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradley D. E., Pitt T. L. Pilus-dependence of four Pseudomonas aeruginosa bacteriophages with non-contractile tails. J Gen Virol. 1974 Jul;24(1):1–15. doi: 10.1099/0022-1317-24-1-1. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Doig P., Todd T., Sastry P. A., Lee K. K., Hodges R. S., Paranchych W., Irvin R. T. Role of pili in adhesion of Pseudomonas aeruginosa to human respiratory epithelial cells. Infect Immun. 1988 Jun;56(6):1641–1646. doi: 10.1128/iai.56.6.1641-1646.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dunn D. L., Barke R. A., Knight N. B., Humphrey E. W., Simmons R. L. Role of resident macrophages, peripheral neutrophils, and translymphatic absorption in bacterial clearance from the peritoneal cavity. Infect Immun. 1985 Aug;49(2):257–264. doi: 10.1128/iai.49.2.257-264.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Friedman A. M., Long S. R., Brown S. E., Buikema W. J., Ausubel F. M. Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene. 1982 Jun;18(3):289–296. doi: 10.1016/0378-1119(82)90167-6. [DOI] [PubMed] [Google Scholar]
  9. Frost L. S., Paranchych W. Composition and molecular weight of pili purified from Pseudomonas aeruginosa K. J Bacteriol. 1977 Jul;131(1):259–269. doi: 10.1128/jb.131.1.259-269.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hancock R. E., Carey A. M. Outer membrane of Pseudomonas aeruginosa: heat- 2-mercaptoethanol-modifiable proteins. J Bacteriol. 1979 Dec;140(3):902–910. doi: 10.1128/jb.140.3.902-910.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kelly N. M., Battershill J. L., Kuo S., Arbuthnott J. P., Hancock R. E. Colonial dissociation and susceptibility to phagocytosis of Pseudomonas aeruginosa grown in a chamber implant model in mice. Infect Immun. 1987 Nov;55(11):2841–2843. doi: 10.1128/iai.55.11.2841-2843.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kelly N. M., Bell A., Hancock R. E. Surface characteristics of Pseudomonas aeruginosa grown in a chamber implant model in mice and rats. Infect Immun. 1989 Feb;57(2):344–350. doi: 10.1128/iai.57.2.344-350.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kluftinger J. L., Kelly N. M., Hancock R. E. Stimulation by fibronectin of macrophage-mediated phagocytosis of Pseudomonas aeruginosa. Infect Immun. 1989 Mar;57(3):817–822. doi: 10.1128/iai.57.3.817-822.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kluftinger J. L., Kelly N. M., Jost B. H., Hancock R. E. Fibronectin as an enhancer of nonopsonic phagocytosis of Pseudomonas aeruginosa by macrophages. Infect Immun. 1989 Sep;57(9):2782–2785. doi: 10.1128/iai.57.9.2782-2785.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Paranchych W., Sastry P. A., Frost L. S., Carpenter M., Armstrong G. D., Watts T. H. Biochemical studies on pili isolated from Pseudomonas aeruginosa strain PAO. Can J Microbiol. 1979 Oct;25(10):1175–1181. doi: 10.1139/m79-182. [DOI] [PubMed] [Google Scholar]
  16. Paranchych W., Sastry P. A., Volpel K., Loh B. A., Speert D. P. Fimbriae (pili): molecular basis of Pseudomonas aeruginosa adherence. Clin Invest Med. 1986;9(2):113–118. [PubMed] [Google Scholar]
  17. Pasloske B. L., Finlay B. B., Paranchych W. Cloning and sequencing of the Pseudomonas aeruginosa PAK pilin gene. FEBS Lett. 1985 Apr 22;183(2):408–412. doi: 10.1016/0014-5793(85)80821-8. [DOI] [PubMed] [Google Scholar]
  18. Pasloske B. L., Paranchych W. The expression of mutant pilins in Pseudomonas aeruginosa: fifth position glutamate affects pilin methylation. Mol Microbiol. 1988 Jul;2(4):489–495. doi: 10.1111/j.1365-2958.1988.tb00055.x. [DOI] [PubMed] [Google Scholar]
  19. Reynolds H. Y., Kazmierowski J. A., Newball H. H. Specificity of opsonic antibodies to enhance phagocytosis of Pseudomonas aeruginosa by human alveolar macrophages. J Clin Invest. 1975 Aug;56(2):376–385. doi: 10.1172/JCI108102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Robinson M. K., Bennett P. M., Falkow S., Dodd H. M. Isolation of a temperature-sensitive derivative of RP1. Plasmid. 1980 May;3(3):343–347. doi: 10.1016/0147-619x(80)90047-5. [DOI] [PubMed] [Google Scholar]
  21. Ruvkun G. B., Ausubel F. M. A general method for site-directed mutagenesis in prokaryotes. Nature. 1981 Jan 1;289(5793):85–88. doi: 10.1038/289085a0. [DOI] [PubMed] [Google Scholar]
  22. Sastry P. A., Finlay B. B., Pasloske B. L., Paranchych W., Pearlstone J. R., Smillie L. B. Comparative studies of the amino acid and nucleotide sequences of pilin derived from Pseudomonas aeruginosa PAK and PAO. J Bacteriol. 1985 Nov;164(2):571–577. doi: 10.1128/jb.164.2.571-577.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sato H., Okinaga K. Role of pili in the adherence of Pseudomonas aeruginosa to mouse epidermal cells. Infect Immun. 1987 Aug;55(8):1774–1778. doi: 10.1128/iai.55.8.1774-1778.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schaffner T., Keller H. U., Hess M. W., Cottier H. Macrophage functions in antimicrobial defense. Klin Wochenschr. 1982 Jul 15;60(14):720–726. doi: 10.1007/BF01716563. [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., Wright S. D., Silverstein S. C., Mah B. Functional characterization of macrophage receptors for in vitro phagocytosis of unopsonized Pseudomonas aeruginosa. J Clin Invest. 1988 Sep;82(3):872–879. doi: 10.1172/JCI113692. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  28. Woods D. E., Straus D. C., Johanson W. G., Jr, Berry V. K., Bass J. A. Role of pili in adherence of Pseudomonas aeruginosa to mammalian buccal epithelial cells. Infect Immun. 1980 Sep;29(3):1146–1151. doi: 10.1128/iai.29.3.1146-1151.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wright S. D., Licht M. R., Craigmyle L. S., Silverstein S. C. Communication between receptors for different ligands on a single cell: ligation of fibronectin receptors induces a reversible alteration in the function of complement receptors on cultured human monocytes. J Cell Biol. 1984 Jul;99(1 Pt 1):336–339. doi: 10.1083/jcb.99.1.336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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