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. 1994 Feb;62(2):596–605. doi: 10.1128/iai.62.2.596-605.1994

Nonmotility and phagocytic resistance of Pseudomonas aeruginosa isolates from chronically colonized patients with cystic fibrosis.

E Mahenthiralingam 1, M E Campbell 1, D P Speert 1
PMCID: PMC186146  PMID: 8300217

Abstract

Although Pseudomonas aeruginosa chronically colonizes most older patients with cystic fibrosis (CF), bacterial features responsible for its persistence are understood poorly. We observed that many P. aeruginosa isolates from chronically colonized patients were nonmotile and resistant to phagocytosis by macrophages. P. aeruginosa isolates were collected from 20 CF patients for up to 10 years. Isolates from early colonization were highly motile and expressed both flagellin and pilin. However, many isolates from chronically colonized patients lacked flagellin expression and were nonmotile; a total of 1,030 P. aeruginosa CF isolates were examined, of which 39% were nonmotile. Moreover, sequential isolates recovered from several of the CF patients were consistently nonmotile for up to 10 years. Lack of motility was rare among environmental isolates (1.4%) and other clinical isolates (3.7%) of P. aeruginosa examined. Partial complementation of motility in nonmotile P. aeruginosa isolates was achieved by introduction of extra copies of the rpoN locus carried on plasmid pPT212, indicating that the alternate sigma factor, RpoN, may be involved in the coordinate regulation of virulence factors during CF infection. We hypothesize that the nonmotile phenotype may provide P. aeruginosa a survival advantage in chronic CF infection by enabling it to resist phagocytosis and conserve energy.

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Selected References

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  1. Baltimore R. S., Shedd D. G. The role of complement in the opsonization of mucoid and non-mucoid strains of Pseudomonas aeruginosa. Pediatr Res. 1983 Dec;17(12):952–958. doi: 10.1203/00006450-198312000-00006. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Burke V., Robinson J. O., Richardson C. J., Bundell C. S. Longitudinal studies of virulence factors of Pseudomonas aeruginosa in cystic fibrosis. Pathology. 1991 Apr;23(2):145–148. doi: 10.3109/00313029109060814. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Campbell M. E., Farmer S. W., Speert D. P. New selective medium for Pseudomonas aeruginosa with phenanthroline and 9-chloro-9-[4-(diethylamino)phenyl]-9,10-dihydro-10- phenylacridine hydrochloride (C-390). J Clin Microbiol. 1988 Sep;26(9):1910–1912. doi: 10.1128/jcm.26.9.1910-1912.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chi E., Mehl T., Nunn D., Lory S. Interaction of Pseudomonas aeruginosa with A549 pneumocyte cells. Infect Immun. 1991 Mar;59(3):822–828. doi: 10.1128/iai.59.3.822-828.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Drake D., Montie T. C. Flagella, motility and invasive virulence of Pseudomonas aeruginosa. J Gen Microbiol. 1988 Jan;134(1):43–52. doi: 10.1099/00221287-134-1-43. [DOI] [PubMed] [Google Scholar]
  8. Figurski D. H., Helinski D. R. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Hancock R. E., Poole K., Benz R. Outer membrane protein P of Pseudomonas aeruginosa: regulation by phosphate deficiency and formation of small anion-specific channels in lipid bilayer membranes. J Bacteriol. 1982 May;150(2):730–738. doi: 10.1128/jb.150.2.730-738.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hazlett L. D., Moon M. M., Singh A., Berk R. S., Rudner X. L. Analysis of adhesion, piliation, protease production and ocular infectivity of several P. aeruginosa strains. Curr Eye Res. 1991 Apr;10(4):351–362. doi: 10.3109/02713689108996341. [DOI] [PubMed] [Google Scholar]
  12. Høiby N., Rosendal K. Epidemiology of Pseudomonas aeruginosa infection in patients treated at a cystic fibrosis centre. Acta Pathol Microbiol Scand B. 1980 Jun;88(3):125–131. doi: 10.1111/j.1699-0463.1980.tb02617.x. [DOI] [PubMed] [Google Scholar]
  13. Kelly N. M., Kluftinger J. L., Pasloske B. L., Paranchych W., Hancock R. E. Pseudomonas aeruginosa pili as ligands for nonopsonic phagocytosis by fibronectin-stimulated macrophages. Infect Immun. 1989 Dec;57(12):3841–3845. doi: 10.1128/iai.57.12.3841-3845.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Krieg D. P., Helmke R. J., German V. F., Mangos J. A. Resistance of mucoid Pseudomonas aeruginosa to nonopsonic phagocytosis by alveolar macrophages in vitro. Infect Immun. 1988 Dec;56(12):3173–3179. doi: 10.1128/iai.56.12.3173-3179.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kyhse-Andersen J. Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods. 1984 Dec;10(3-4):203–209. doi: 10.1016/0165-022x(84)90040-x. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Li C., Louise C. J., Shi W., Adler J. Adverse conditions which cause lack of flagella in Escherichia coli. J Bacteriol. 1993 Apr;175(8):2229–2235. doi: 10.1128/jb.175.8.2229-2235.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Luzar M. A., Montie T. C. Avirulence and altered physiological properties of cystic fibrosis strains of Pseudomonas aeruginosa. Infect Immun. 1985 Nov;50(2):572–576. doi: 10.1128/iai.50.2.572-576.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Luzar M. A., Thomassen M. J., Montie T. C. Flagella and motility alterations in Pseudomonas aeruginosa strains from patients with cystic fibrosis: relationship to patient clinical condition. Infect Immun. 1985 Nov;50(2):577–582. doi: 10.1128/iai.50.2.577-582.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mohr C. D., Martin D. W., Konyecsni W. M., Govan J. R., Lory S., Deretic V. Role of the far-upstream sites of the algD promoter and the algR and rpoN genes in environmental modulation of mucoidy in Pseudomonas aeruginosa. J Bacteriol. 1990 Nov;172(11):6576–6580. doi: 10.1128/jb.172.11.6576-6580.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mork T., Hancock R. E. Mechanisms of nonopsonic phagocytosis of Pseudomonas aeruginosa. Infect Immun. 1993 Aug;61(8):3287–3293. doi: 10.1128/iai.61.8.3287-3293.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nelson J. W., Tredgett M. W., Sheehan J. K., Thornton D. J., Notman D., Govan J. R. Mucinophilic and chemotactic properties of Pseudomonas aeruginosa in relation to pulmonary colonization in cystic fibrosis. Infect Immun. 1990 Jun;58(6):1489–1495. doi: 10.1128/iai.58.6.1489-1495.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pasloske B. L., Joffe A. M., Sun Q., Volpel K., Paranchych W., Eftekhar F., Speert D. P. Serial isolates of Pseudomonas aeruginosa from a cystic fibrosis patient have identical pilin sequences. Infect Immun. 1988 Mar;56(3):665–672. doi: 10.1128/iai.56.3.665-672.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pier G. B., Meluleni G., Neuger E. A murine model of chronic mucosal colonization by Pseudomonas aeruginosa. Infect Immun. 1992 Nov;60(11):4768–4776. doi: 10.1128/iai.60.11.4768-4776.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Saiman L., Ishimoto K., Lory S., Prince A. The effect of piliation and exoproduct expression on the adherence of Pseudomonas aeruginosa to respiratory epithelial monolayers. J Infect Dis. 1990 Mar;161(3):541–548. doi: 10.1093/infdis/161.3.541. [DOI] [PubMed] [Google Scholar]
  26. Shi W., Li C., Louise C. J., Adler J. Mechanism of adverse conditions causing lack of flagella in Escherichia coli. J Bacteriol. 1993 Apr;175(8):2236–2240. doi: 10.1128/jb.175.8.2236-2240.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Simpson D. A., Ramphal R., Lory S. Genetic analysis of Pseudomonas aeruginosa adherence: distinct genetic loci control attachment to epithelial cells and mucins. Infect Immun. 1992 Sep;60(9):3771–3779. doi: 10.1128/iai.60.9.3771-3779.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  29. Smith R. J., Iden S. S. Properties of calcium ionophore-induced generation of superoxide anion by human neutrophils. Inflammation. 1981 Sep;5(3):177–192. doi: 10.1007/BF00914442. [DOI] [PubMed] [Google Scholar]
  30. Speert D. P., Eftekhar F., Puterman M. L. Nonopsonic phagocytosis of strains of Pseudomonas aeruginosa from cystic fibrosis patients. Infect Immun. 1984 Mar;43(3):1006–1011. doi: 10.1128/iai.43.3.1006-1011.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Speert D. P., Gordon S. Phagocytosis of unopsonized Pseudomonas aeruginosa by murine macrophages is a two-step process requiring glucose. J Clin Invest. 1992 Sep;90(3):1085–1092. doi: 10.1172/JCI115924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. Starnbach M. N., Lory S. The fliA (rpoF) gene of Pseudomonas aeruginosa encodes an alternative sigma factor required for flagellin synthesis. Mol Microbiol. 1992 Feb;6(4):459–469. doi: 10.1111/j.1365-2958.1992.tb01490.x. [DOI] [PubMed] [Google Scholar]
  35. Totten P. A., Lara J. C., Lory S. The rpoN gene product of Pseudomonas aeruginosa is required for expression of diverse genes, including the flagellin gene. J Bacteriol. 1990 Jan;172(1):389–396. doi: 10.1128/jb.172.1.389-396.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Valeyre D., Soler P., Basset G., Loiseau P., Pre J., Turbie P., Battesti J. P., Georges R. Glucose, K+, and albumin concentrations in the alveolar milieu of normal humans and pulmonary sarcoidosis patients. Am Rev Respir Dis. 1991 May;143(5 Pt 1):1096–1101. doi: 10.1164/ajrccm/143.5_Pt_1.1096. [DOI] [PubMed] [Google Scholar]
  37. WAHBA A. H., DARRELL J. H. THE IDENTIFICATION OF ATYPICAL STRAINS OF PSEUDOMONAS AERUGINOSA. J Gen Microbiol. 1965 Mar;38:329–342. doi: 10.1099/00221287-38-3-329. [DOI] [PubMed] [Google Scholar]

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