Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1987 Oct;55(10):2420–2427. doi: 10.1128/iai.55.10.2420-2427.1987

Nonoxidative antimicrobial effects of human polymorphonuclear leukocyte granule proteins on Chlamydia spp. in vitro.

K B Register 1, C H Davis 1, P B Wyrick 1, W M Shafer 1, J K Spitznagel 1
PMCID: PMC260724  PMID: 3653985

Abstract

Proteins from isolated granules of human polymorphonuclear leukocytes were assessed for their nonoxidative microbicidal effect on chlamydiae by two different methods: a radioisotope assay for elementary body integrity and a biological assay for inclusion development. Crude granule extract, which consisted of a mixture of all granule proteins, caused a 20 to 30% decrease in infectivity and a 52% decrease in infectivity when incubated with Chlamydia psittaci CAL-10 and Chlamydia trachomatis serovar E, respectively. To define more specifically the components that were damaging to chlamydiae, crude granule extract was subjected to Sephadex G-75 column chromatography and isolated granule fractions were obtained. Only fractions containing lysozyme as the major component consistently caused reductions in infectivity of C. trachomatis elementary bodies. In contrast, fractions collected after the lysozyme fraction, containing proteins with molecular masses of 13,000 daltons or less, had detrimental effects on C. psittaci infectivity. Additional experiments using highly purified human polymorphonuclear leukocyte lysozyme confirmed its infectivity-reducing action upon C. trachomatis but not upon C. psittaci.

Full text

PDF
2422

Images in this article

2421Image
on p.2421
2424Image
on p.2424

Selected References

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

  1. Barbour A. G., Amano K., Hackstadt T., Perry L., Caldwell H. D. Chlamydia trachomatis has penicillin-binding proteins but not detectable muramic acid. J Bacteriol. 1982 Jul;151(1):420–428. doi: 10.1128/jb.151.1.420-428.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barenfanger J., MacDonald A. B. The role of immunoglobulin in the neutralization of trachoma infectivity. J Immunol. 1974 Nov;113(5):1607–1617. [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Caldwell H. D., Hitchcock P. J. Monoclonal antibody against a genus-specific antigen of Chlamydia species: location of the epitope on chlamydial lipopolysaccharide. Infect Immun. 1984 May;44(2):306–314. doi: 10.1128/iai.44.2.306-314.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cech P., Lehrer R. I. Phagolysosomal pH of human neutrophils. Blood. 1984 Jan;63(1):88–95. [PubMed] [Google Scholar]
  6. Cummins C. S., Stimpson S. A., Tuttle R. L., Weck C. J. Observations on the nature of the determinants in anaerobic coryneforms which produce splenomegaly. J Reticuloendothel Soc. 1981 Jan;29(1):1–13. [PubMed] [Google Scholar]
  7. Eissenberg L. G., Wyrick P. B., Davis C. H., Rumpp J. W. Chlamydia psittaci elementary body envelopes: ingestion and inhibition of phagolysosome fusion. Infect Immun. 1983 May;40(2):741–751. doi: 10.1128/iai.40.2.741-751.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Evans B. A. Chlamydial infection of the human cervix--an ultrastructural study. J Infect. 1982 May;4(3):225–228. doi: 10.1016/s0163-4453(82)92479-3. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Garrett A. J., Harrison M. J., Manire G. P. A search for the bacterial mucopeptide component, muramic acid, in Chlamydia. J Gen Microbiol. 1974 Jan;80(1):315–318. doi: 10.1099/00221287-80-1-315. [DOI] [PubMed] [Google Scholar]
  11. Hatch T. P., Allan I., Pearce J. H. Structural and polypeptide differences between envelopes of infective and reproductive life cycle forms of Chlamydia spp. J Bacteriol. 1984 Jan;157(1):13–20. doi: 10.1128/jb.157.1.13-20.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kondo L. R., Hanna L., Keshishyan H. Reduction in chlamydial infectivity by lysozyme. Proc Soc Exp Biol Med. 1973 Jan;142(1):131–132. doi: 10.3181/00379727-142-36974. [DOI] [PubMed] [Google Scholar]
  13. Kramer M. J., Gordon F. B. Ultrastructural analysis of the effects of penicillin and chlortetracycline on the development of a genital tract Chlamydia. Infect Immun. 1971 Feb;3(2):333–341. doi: 10.1128/iai.3.2.333-341.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kuo C. C. Cultures of Chlamydia trachomatis in mouse peritoneal macrophages: factors affecting organism growth. Infect Immun. 1978 May;20(2):439–445. doi: 10.1128/iai.20.2.439-445.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kuo C. C., Grayston T. Interaction of Chlamydia trachomatis organisms and HeLa 229 cells. Infect Immun. 1976 Apr;13(4):1103–1109. doi: 10.1128/iai.13.4.1103-1109.1976. [DOI] [PMC free article] [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. Laible N. J., Germaine G. R. Bactericidal activity of human lysozyme, muramidase-inactive lysozyme, and cationic polypeptides against Streptococcus sanguis and Streptococcus faecalis: inhibition by chitin oligosaccharides. Infect Immun. 1985 Jun;48(3):720–728. doi: 10.1128/iai.48.3.720-728.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Matsumoto A., Manire G. P. Electron microscopic observations on the effects of penicillin on the morphology of Chlamydia psittaci. J Bacteriol. 1970 Jan;101(1):278–285. doi: 10.1128/jb.101.1.278-285.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Megran D. W., Stiver H. G., Bowie W. R. Complement activation and stimulation of chemotaxis by Chlamydia trachomatis. Infect Immun. 1985 Sep;49(3):670–673. doi: 10.1128/iai.49.3.670-673.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Modrzakowski M. C., Paranavitana C. M. Bactericidal activity of fractionated granule contents from human polymorphonuclear leukocytes: role of bacterial membrane lipid. Infect Immun. 1981 May;32(2):668–674. doi: 10.1128/iai.32.2.668-674.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Modrzakowski M. C., Spitznagel J. K. Bactericidal activity of fractionated granule contents from human polymorphonuclear leukocytes: antagonism of granule cationic proteins by lipopolysaccharide. Infect Immun. 1979 Aug;25(2):597–602. doi: 10.1128/iai.25.2.597-602.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mårdh P. A., Colleen S., Sylwan J. Inhibitory effect on the formation of chlamydial inclusions in McCoy cells by seminal fluid and some of its components. Invest Urol. 1980 May;17(6):510–513. [PubMed] [Google Scholar]
  23. Narita T., Manire G. P. Protein-carbohydrate-lipid complex isolated from the cell envelopes of Chlamydia psittaci in alkaline buffer and ethylenediaminetetraacetate. J Bacteriol. 1976 Jan;125(1):308–316. doi: 10.1128/jb.125.1.308-316.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Narita T., Wyrick P. B., Manire G. P. Effect of alkali on the structure of cell envelopes of Chlamydia psittaci elementary bodies. J Bacteriol. 1976 Jan;125(1):300–307. doi: 10.1128/jb.125.1.300-307.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Newhall W. J., Batteiger B., Jones R. B. Analysis of the human serological response to proteins of Chlamydia trachomatis. Infect Immun. 1982 Dec;38(3):1181–1189. doi: 10.1128/iai.38.3.1181-1189.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Newhall W. J., Jones R. B. Disulfide-linked oligomers of the major outer membrane protein of chlamydiae. J Bacteriol. 1983 May;154(2):998–1001. doi: 10.1128/jb.154.2.998-1001.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nurminen M., Leinonen M., Saikku P., Mäkelä P. H. The genus-specific antigen of Chlamydia: resemblance to the lipopolysaccharide of enteric bacteria. Science. 1983 Jun 17;220(4603):1279–1281. doi: 10.1126/science.6344216. [DOI] [PubMed] [Google Scholar]
  28. Odeberg H., Olsson I. Microbicidal mechanisms of human granulocytes: synergistic effects of granulocyte elastase and myeloperoxidase or chymotrypsin-like cationic protein. Infect Immun. 1976 Dec;14(6):1276–1283. doi: 10.1128/iai.14.6.1276-1283.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Osserman E. F., Lawlor D. P. Serum and urinary lysozyme (muramidase) in monocytic and monomyelocytic leukemia. J Exp Med. 1966 Nov 1;124(5):921–952. doi: 10.1084/jem.124.5.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. PERKINS H. R., ALLISON A. C. Cell-wall constituents of rickettsiae and psittacosis-lymphogranuloma organisms. J Gen Microbiol. 1963 Mar;30:469–480. doi: 10.1099/00221287-30-3-469. [DOI] [PubMed] [Google Scholar]
  31. Pryzwansky K. B., Martin L. E., Spitznagel J. K. Immunocytochemical localization of myeloperoxidase, lactoferrin, lysozyme and neutral proteases in human monocytes and neutrophilic granulocytes. J Reticuloendothel Soc. 1978 Sep;24(3):295–310. [PubMed] [Google Scholar]
  32. Register K. B., Morgan P. A., Wyrick P. B. Interaction between Chlamydia spp. and human polymorphonuclear leukocytes in vitro. Infect Immun. 1986 Jun;52(3):664–670. doi: 10.1128/iai.52.3.664-670.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rosenthal R. S., Folkening W. J., Miller D. R., Swim S. C. Resistance of O-acetylated gonococcal peptidoglycan to human peptidoglycan-degrading enzymes. Infect Immun. 1983 Jun;40(3):903–911. doi: 10.1128/iai.40.3.903-911.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. SHUGAR D. The measurement of lysozyme activity and the ultra-violet inactivation of lysozyme. Biochim Biophys Acta. 1952 Mar;8(3):302–309. doi: 10.1016/0006-3002(52)90045-0. [DOI] [PubMed] [Google Scholar]
  35. Segal A. W., Geisow M., Garcia R., Harper A., Miller R. The respiratory burst of phagocytic cells is associated with a rise in vacuolar pH. Nature. 1981 Apr 2;290(5805):406–409. doi: 10.1038/290406a0. [DOI] [PubMed] [Google Scholar]
  36. Selsted M. E., Harwig S. S., Ganz T., Schilling J. W., Lehrer R. I. Primary structures of three human neutrophil defensins. J Clin Invest. 1985 Oct;76(4):1436–1439. doi: 10.1172/JCI112121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Selsted M. E., Szklarek D., Ganz T., Lehrer R. I. Activity of rabbit leukocyte peptides against Candida albicans. Infect Immun. 1985 Jul;49(1):202–206. doi: 10.1128/iai.49.1.202-206.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Shafer W. M., Martin L. E., Spitznagel J. K. Cationic antimicrobial proteins isolated from human neutrophil granulocytes in the presence of diisopropyl fluorophosphate. Infect Immun. 1984 Jul;45(1):29–35. doi: 10.1128/iai.45.1.29-35.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shafer W. M., Martin L. E., Spitznagel J. K. Late intraphagosomal hydrogen ion concentration favors the in vitro antimicrobial capacity of a 37-kilodalton cationic granule protein of human neutrophil granulocytes. Infect Immun. 1986 Sep;53(3):651–655. doi: 10.1128/iai.53.3.651-655.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Shafer W. M., Onunka V., Hitchcock P. J. A spontaneous mutant of Neisseria gonorrhoeae with decreased resistance to neutrophil granule proteins. J Infect Dis. 1986 May;153(5):910–917. doi: 10.1093/infdis/153.5.910. [DOI] [PubMed] [Google Scholar]
  41. Spitznagel J. K. Nonoxidative antimicrobial reactions of leukocytes. Contemp Top Immunobiol. 1984;14:283–343. doi: 10.1007/978-1-4757-4862-8_10. [DOI] [PubMed] [Google Scholar]
  42. Starkey P. M., Barrett A. J. Human cathepsin G. Catalytic and immunological properties. Biochem J. 1976 May 1;155(2):273–278. doi: 10.1042/bj1550273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Starkey P. M., Barrett A. J. Human lysosomal elastase. Catalytic and immunological properties. Biochem J. 1976 May 1;155(2):265–271. doi: 10.1042/bj1550265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Stimpson S. A., Lerch R. A., Cleland D. R., Yarnall D. P., Clark R. L., Cromartie W. J., Schwab J. H. Effect of acetylation on arthropathic activity of group A streptococcal peptidoglycan-polysaccharide fragments. Infect Immun. 1987 Jan;55(1):16–23. doi: 10.1128/iai.55.1.16-23.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. TAMURA A., HIGASHI N. PURIFICATION AND CHEMICAL COMPOSITION OF MENINGOPNEUMONITIS VIRUS. Virology. 1963 Aug;20:596–604. doi: 10.1016/0042-6822(63)90284-8. [DOI] [PubMed] [Google Scholar]
  46. Vasstrand E. N., Jensen H. B. Affinity chromatography of human saliva lysozyme and effect of pH and ionic strength on lytic activity. Scand J Dent Res. 1980 Jun;88(3):219–228. doi: 10.1111/j.1600-0722.1980.tb01218.x. [DOI] [PubMed] [Google Scholar]
  47. Ward M. E. Chlamydial classification, development and structure. Br Med Bull. 1983 Apr;39(2):109–115. doi: 10.1093/oxfordjournals.bmb.a071800. [DOI] [PubMed] [Google Scholar]
  48. 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]
  49. Weiss J., Hutzler M., Kao L. Environmental modulation of lipopolysaccharide chain length alters the sensitivity of Escherichia coli to the neutrophil bactericidal/permeability-increasing protein. Infect Immun. 1986 Feb;51(2):594–599. doi: 10.1128/iai.51.2.594-599.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Wyrick P. B., Brownridge E. A. Growth of Chlamydia psittaci in macrophages. Infect Immun. 1978 Mar;19(3):1054–1060. doi: 10.1128/iai.19.3.1054-1060.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wyrick P. B., Brownridge E. A., Ivins B. E. Interaction of Chlamydia psittaci with mouse peritoneal macrophages. Infect Immun. 1978 Mar;19(3):1061–1067. doi: 10.1128/iai.19.3.1061-1067.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Yong E. C., Chi E. Y., Chen W. J., Kuo C. C. Degradation of Chlamydia trachomatis in human polymorphonuclear leukocytes: an ultrastructural study of peroxidase-positive phagolysosomes. Infect Immun. 1986 Aug;53(2):427–431. doi: 10.1128/iai.53.2.427-431.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Yong E. C., Klebanoff S. J., Kuo C. C. Toxic effect of human polymorphonuclear leukocytes on Chlamydia trachomatis. Infect Immun. 1982 Aug;37(2):422–426. doi: 10.1128/iai.37.2.422-426.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Yoshimoto T., Tobiishi M., Tsuru D. Affinity chromatographic purification of human lysozyme, with special reference to human leukemia lysozyme. J Biochem. 1976 Oct;80(4):703–709. doi: 10.1093/oxfordjournals.jbchem.a131329. [DOI] [PubMed] [Google Scholar]

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

RESOURCES