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. 1988 Mar;56(3):552–556. doi: 10.1128/iai.56.3.552-556.1988

Antimicrobial mechanisms against Acinetobacter calcoaceticus of rat polymorphonuclear leukocyte granule extract.

M J Loeffelholz 1, M C Modrzakowski 1
PMCID: PMC259325  PMID: 2449397

Abstract

The antimicrobial mechanisms of rat polymorphonuclear leukocyte granule extract and isolated extract fractions against Acinetobacter calcoaceticus were examined. Crude granule extract and a fraction containing low-molecular-weight cationic peptides (peak D) reduced the viability of A. calcoaceticus and inhibited the uptake of radiolabeled macromolecule precursors by cells. The inhibitory activity observed with peak D was not as great as that of crude granule extract containing equivalent amounts of peak D protein. Crude extract also inhibited incorporation of uracil into trichloroacetic acid-precipitable material, while no isolated fraction, including peak D, had any substantial effect on incorporation. The antimicrobial activities of crude granule extract were more sensitive to boiling than those of isolated peak D. Preincubation of A. calcoaceticus with either crude granule extract or a fraction (peak B) possessing proteolytic activity but lacking any antimicrobial activity caused cells to become sensitive to a subinhibitory concentration of actinomycin D, suggesting that granule extract and peak B increase the outer membrane permeability of A. calcoaceticus. The antimicrobial granule extract fraction, peak D, did not affect outer membrane permeability. These results suggest that rat polymorphonuclear leukocyte granule extract reduces the viability of A. calcoaceticus by inhibiting the transport and incorporation of macromolecule precursors and that either whole granule extract is required for complete antimicrobial activity or an unidentified component is responsible for antimicrobial activity in addition to peak D. The granule extract activity that increases outer membrane permeability does not appear to be directly responsible for the observed decrease in viability.

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Selected References

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

  1. Bergogne-Berezin E., Joly-Guillou M. L. An underestimated nosocomial pathogen, Acinetobacter calcoaceticus. J Antimicrob Chemother. 1985 Nov;16(5):535–538. doi: 10.1093/jac/16.5.535. [DOI] [PubMed] [Google Scholar]
  2. Buck P., Rest R. F. Effects of human neutrophil granule extracts on macromolecular synthesis in Neisseria gonorrhoeae. Infect Immun. 1981 Aug;33(2):426–433. doi: 10.1128/iai.33.2.426-433.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Elsbach P., Beckerdite S., Pettis P., Franson R. Persistence of regulation of macromolecular synthesis by Escherichia coli during killing by disrupted rabbit granulocytes. Infect Immun. 1974 Apr;9(4):663–668. doi: 10.1128/iai.9.4.663-668.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ganz T., Selsted M. E., Szklarek D., Harwig S. S., Daher K., Bainton D. F., Lehrer R. I. Defensins. Natural peptide antibiotics of human neutrophils. J Clin Invest. 1985 Oct;76(4):1427–1435. doi: 10.1172/JCI112120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Glew R. H., Moellering R. C., Jr, Kunz L. J. Infections with Acinetobacter calcoaceticus (Herellea vaginicola): clinical and laboratory studies. Medicine (Baltimore) 1977 Mar;56(2):79–97. doi: 10.1097/00005792-197703000-00001. [DOI] [PubMed] [Google Scholar]
  6. Hodinka R. L., Modrzakowski M. C. Bactericidal activity of granule contents from rat polymorphonuclear leukocytes. Infect Immun. 1983 Apr;40(1):139–146. doi: 10.1128/iai.40.1.139-146.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hodinka R. L., Modrzakowski M. C. Granule contents from rat polymorphonuclear leukocytes: antimicrobial properties and characterization. Can J Microbiol. 1986 Jun;32(6):498–504. doi: 10.1139/m86-091. [DOI] [PubMed] [Google Scholar]
  8. Hovde C. J., Gray B. H. Physiological effects of a bactericidal protein from human polymorphonuclear leukocytes on Pseudomonas aeruginosa. Infect Immun. 1986 Apr;52(1):90–95. doi: 10.1128/iai.52.1.90-95.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lehrer R. I., Szklarek D., Ganz T., Selsted M. E. Synergistic activity of rabbit granulocyte peptides against Candida albicans. Infect Immun. 1986 Jun;52(3):902–904. doi: 10.1128/iai.52.3.902-904.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Loeffelholz M. J., Modrzakowski M. C. Isolation of cationic peptides from rat polymorphonuclear leukocyte granule contents using fast protein liquid chromatography. Anal Biochem. 1986 Nov 1;158(2):377–381. doi: 10.1016/0003-2697(86)90564-6. [DOI] [PubMed] [Google Scholar]
  11. Loeffelholz M. J., Modrzakowski M. C. Outer membrane permeability of Acinetobacter calcoaceticus mediates susceptibility to rat polymorphonuclear leukocyte granule contents. Infect Immun. 1987 Sep;55(9):2296–2299. doi: 10.1128/iai.55.9.2296-2299.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Loeffelholz M. J., Modrzakowski M. C. Plasmid RP1-mediated susceptibility of Acinetobacter calcoaceticus to rat polymorphonuclear leukocyte granule contents. Infect Immun. 1986 Dec;54(3):705–709. doi: 10.1128/iai.54.3.705-709.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nikaido H., Nakae T. The outer membrane of Gram-negative bacteria. Adv Microb Physiol. 1979;20:163–250. doi: 10.1016/s0065-2911(08)60208-8. [DOI] [PubMed] [Google Scholar]
  14. Odeberg H., Olsson I. Mechanisms for the microbicidal activity of cationic proteins of human granulocytes. Infect Immun. 1976 Dec;14(6):1269–1275. doi: 10.1128/iai.14.6.1269-1275.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Selsted M. E., Szklarek D., Lehrer R. I. Purification and antibacterial activity of antimicrobial peptides of rabbit granulocytes. Infect Immun. 1984 Jul;45(1):150–154. doi: 10.1128/iai.45.1.150-154.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shafer W. M., Onunka V. C., Martin L. E. Antigonococcal activity of human neutrophil cathepsin G. Infect Immun. 1986 Oct;54(1):184–188. doi: 10.1128/iai.54.1.184-188.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Van Houte A. J., Elsbach P., Verkleij A., Weiss J. Killing of Escherichia coli by a granulocyte fraction occurs without recognizable ultrastructural alterations in the bacterial envelope, as studied by freeze-fracture electron microscopy. Infect Immun. 1977 Feb;15(2):556–559. doi: 10.1128/iai.15.2.556-559.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Weiss J., Elsbach P., Olsson I., Odeberg H. Purification and characterization of a potent bactericidal and membrane active protein from the granules of human polymorphonuclear leukocytes. J Biol Chem. 1978 Apr 25;253(8):2664–2672. [PubMed] [Google Scholar]
  19. Weiss J., Victor M., Stendhal O., Elsbach P. Killing of gram-negative bacteria by polymorphonuclear leukocytes: role of an O2-independent bactericidal system. J Clin Invest. 1982 Apr;69(4):959–970. doi: 10.1172/JCI110535. [DOI] [PMC free article] [PubMed] [Google Scholar]

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