1887

Abstract

The uridine incorporation technique for assaying phagocytosis is based on the fact that polymorphonuclear leucocytes are impermeable to labelled uridine, and therefore ingested bacteria inside phagocytic vacuoles will be unable to take it up. Extracellular bacteria, including those adherent to the phagocytic cell surface, can do so however. Differences in uptake between bacteria alone and in the presence of phagocytic cells can be used to measure ingestion. The present paper describes the application of this technique to O-86 as the test organism. It appears that with this test species, the method is unsuccessful, because exposure of the non-ingested bacteria to some soluble product of the triggered polymorphonuclear leucocytes causes a large increase in their uridine uptake rates, over that of the control bacteria. The nature of the product responsible is unknown. It is unconnected with change in the pH of the medium, is heat stable, and is only produced by polymorphonuclear leucocytes which are actively phagocytosing. It may be that a release of phagolysosome contents is responsible.

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1983-12-01
2024-05-03
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References

  1. Beckerdite S., Mooney C., Weiss J., Franson R., Elsbach P. 1974; Early and discrete changes in permeability of Escherichia coli and certain other Gram negative bacteria during killing by granulocytes. Journal of Experimental Medicine 140:396–409
    [Google Scholar]
  2. Boyum A. 1968; Isolation of leukocytes from human blood. Scandinavian Journal of Clinical Investigation 21:1–109
    [Google Scholar]
  3. Braconier J. H., Odeberg H. 1979; Evaluation of phagocytic and bactericidal activities of neutrophil granulocytes: determination of viable extracellular bacteria by their incorporation of 14C-leucine and 3H-thymidine. Scandinavian Journal of Haematology 23:407–415
    [Google Scholar]
  4. Bray G. 1960; A simple efficient liquid scintillator for counting aqueous solutions in a liquid scintillation counter. Annals of Biochemistry 1:279–285
    [Google Scholar]
  5. Bridges C. G., Dasilva G., Yamamura M., Valdimarsson H. 1980; A radiometric assay for the combined measurement of phagocytosis and intracellular killing of Candida albicans. . Clinical and Experimental Immunology 42:226–233
    [Google Scholar]
  6. Dutcher B. S., Reynard A. M., Beck M. E., Cunningham R. K. 1978; Potentiation of antibiotic bactericidal activity by normal human serum. Antimicrobial Agents and Chemotherapy 13:820–826
    [Google Scholar]
  7. Elsbach P., Beckerdite S., Pettis P., Franson R. . 1974; Persistence of regulation of macromolecular synthesis by Escherichia coli during killing by disrupted rabbit granulocytes. Infection and Immunity 9:663–668
    [Google Scholar]
  8. Elsbach P., Weiss J., Franson R. C., Beckerdite S, Harris L. 1979; Separation and purification of a potent bactericidal/perme- ability-increasing protein and a closely associated phospholipase A2 from rabbit polymorphonuclear leukocytes. Journal of Biological Chemistry 254:11000–11009
    [Google Scholar]
  9. Fazeli A. 1982 The ingestion and killing of Escherichia coli by polymorphonuclear leukocytes. Ph.D. thesis University of London.:
    [Google Scholar]
  10. Foroozanfar N., Aghai Z., Ala F., Hobbs J. R. 1976; Inhibition of thymidine uptake by Staphylococci : a new method for the investigation of phagocytosis. Journal of Immunological Methods 11:345–353
    [Google Scholar]
  11. Goldstein I. M. 1976; Polymorphonuclear leukocyte lysosomes and immune tissue injury. Progress in Allergy 20:301–340
    [Google Scholar]
  12. Granelli-Piperno A., Vassali J. D., Beich E. 1977; Secretion of plasminogen activator by human polymorphonuclear leukocytes:modulation by glucocorticoids and other effectors. Journal of Experimental Medicine 146:1693–1706
    [Google Scholar]
  13. Hafeman D. G., Lucas Z. J. 1979; Polymorphonuclear mediated, antibody dependent, cellular cytotoxicity against tumour cells: dependence on oxygen and respiratory burst. Journal of Immunology 123:55–62
    [Google Scholar]
  14. Henson P. M. 1971; The immunologic release of constituents from neutrophil leukocytes. I. The role of antibody and complement on non-phagocytosable surfaces or phagocytosable particles. Journal of Immunology 107:1535–1546
    [Google Scholar]
  15. Johnston R. B., Lehmeyer J. E. 1976; Elaboration of toxic oxygen by-products by neutrophils in a model of immune complex disease. Journal of Clinical Investigation 57:836–841
    [Google Scholar]
  16. Lam C., Mathison G. E. 1979; Phagocytosis measured as inhibition of uridine uptake: a method that distinguishes between surface adherence and ingestion. Journal of Medical Microbiology 12:459–467
    [Google Scholar]
  17. Leffell M. S., Spitznagel J. K. 1975; Fate of human lactoferrin and myeloperoxidase in phagocytosing human neutrophils: effects of immunoglobulin G subclasses and immune complexes coated on latex beads. Infection and Immunity 12:813–820
    [Google Scholar]
  18. Okamura N., Ishibashi S., Takano T. 1979; Evidence for bactericidal activity of polymorphonuclear leukocytes without phagocytosis. Journal of Biochemistry 86:469–475
    [Google Scholar]
  19. Root R. K., Metcalf J., Oshino N., Chance B. 1975; Hydrogen peroxide release from human granulocytes during phagocytosis, I. Documentation, quantitation and some regulation factors. Journal of Clinical Investigation 55:945–955
    [Google Scholar]
  20. Sips H. J., Hamers M. N. 1981; Mechanism of bactericidal action of myeloperoxidase: increased permeability of Escherichia coli cell envelope. Infection and Immunity 31:11–16
    [Google Scholar]
  21. Thong Y. H., Ferrante A. 1977; Phagocytosis and killing of Torulopsis glabrata by human neutrophils. International Review of Science: Medical Science 5:482
    [Google Scholar]
  22. Weiss J., Franson R. C., Schmeider K., Elsbach P. 1976; Reversible envelope effects during and after killing of Escherichia coli W by a highly purified rabbit polymorphonuclear leukocyte fraction. Bio-chimica et biophysica acta 436:154–169
    [Google Scholar]
  23. Wright D. G., Malawista S. E. 1972; The mobilisation and extracellular release of granular enzymes from human leukocytes during phagocytosis. Journal of Cell Biology 53:788–797
    [Google Scholar]
  24. Yamamura M., Boler J., Valdimarsson H. 1977; Phagocytosis measured as inhibition of uridine uptake by Candida albicans. . Journal of Immunological Methods 14:19–24
    [Google Scholar]
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