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. 1977 Sep;17(3):513–520. doi: 10.1128/iai.17.3.513-520.1977

Chemiluminescence Response of Human Leukocytes: Influence of Medium Components on Light Production

Robert D Nelson 1,2, Michael J Herron 1, Jon R Schmidtke 1,2, Richard L Simmons 1,2
PMCID: PMC421154  PMID: 903176

Abstract

Light-producing reactions have been reported to occur after phagocytosis of opsonized particles by human polymorphonuclear neutrophils, eosinophils, and monocytes. Such chemiluminescence appears to be related to the generation of singlet oxygen, superoxide, and hydroxyl radicals, which have also been implicated as microbicidal agents. In examining the influences of various medium components on leukocyte chemiluminescence, we have observed that the amount of light measured is increased by addition of soluble protein, the amino acids tyrosine and tryptophane, or excess zymosan to the reaction medium. These agents appear to produce their effect not by increasing the rate of phagocytosis, but by providing substrate for secondary light-producing reactions. Polystyrene particles do not provide a suitable substrate for such secondary light-producing reactions. This is evidenced by the failure of ingested latex to stimulate high levels of chemiluminescence in the cellular system and their failure to augment light production in two noncellular chemiluminescent reactions. Some of the light generated in the cellular chemiluminescence response may derive from secondary reactions, which occur outside of the phagocyte. Support for this phenomenon is provided by two experiments. In one, addition of supplementary, nonopsonized zymosan to the reaction, after phagocytosis of opsonized zymosan is complete, resulted in an increased level of chemiluminescence. In another, addition of nonopsonized zymosan, together with latex particles, resulted in a significant increase in chemiluminescence. The results of the latter experiment also support our hypothesis that latex does not provide an appropriate substrate for secondary light-producing reactions. These observations suggest that leukocytes activated by phagocytosis generate electronically activated radicals which act intra- and extracellularly and that the amino acids tyrosine and trytophane may provide one substrate through which these agents act.

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Selected References

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

  1. Allen R. C., Loose L. D. Phagocytic activation of a luminol-dependent chemiluminescence in rabbit alveolar and peritoneal macrophages. Biochem Biophys Res Commun. 1976 Mar 8;69(1):245–252. doi: 10.1016/s0006-291x(76)80299-9. [DOI] [PubMed] [Google Scholar]
  2. Allen R. C., Stjernholm R. L., Steele R. H. Evidence for the generation of an electronic excitation state(s) in human polymorphonuclear leukocytes and its participation in bactericidal activity. Biochem Biophys Res Commun. 1972 May 26;47(4):679–684. doi: 10.1016/0006-291x(72)90545-1. [DOI] [PubMed] [Google Scholar]
  3. Allen R. C., Yevich S. J., Orth R. W., Steele R. H. The superoxide anion and singlet molecular oxygen: their role in the microbicidal activity of the polymorphonuclear leukocyte. Biochem Biophys Res Commun. 1974 Oct 8;60(3):909–917. doi: 10.1016/0006-291x(74)90401-x. [DOI] [PubMed] [Google Scholar]
  4. Cheson B. D., Christensen R. L., Sperling R., Kohler B. E., Babior B. M. The origin of the chemiluminescence of phagocytosing granulocytes. J Clin Invest. 1976 Oct;58(4):789–796. doi: 10.1172/JCI108530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  6. Johnston R. B., Jr, Keele B. B., Jr, Misra H. P., Lehmeyer J. E., Webb L. S., Baehner R. L., RaJagopalan K. V. The role of superoxide anion generation in phagocytic bactericidal activity. Studies with normal and chronic granulomatous disease leukocytes. J Clin Invest. 1975 Jun;55(6):1357–1372. doi: 10.1172/JCI108055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Klebanoff S. J. Role of the superoxide anion in the myeloperoxidase-mediated antimicrobial system. J Biol Chem. 1974 Jun 25;249(12):3724–3728. [PubMed] [Google Scholar]
  8. Krinsky N. I. Singlet excited oxygen as a mediator of the antibacterial action of leukocytes. Science. 1974 Oct 25;186(4161):363–365. doi: 10.1126/science.186.4161.363. [DOI] [PubMed] [Google Scholar]
  9. NELKEN D., GILBOA-GARBER N., GUREVITCH J. A method for the simultaneous separation of human thrombocytes and leucocytes. J Clin Pathol. 1960 May;13:266–266. doi: 10.1136/jcp.13.3.266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nelson R. D., Mills E. L., Simmons R. L., Quie P. G. Chemiluminescence response of phagocytizing human monocytes. Infect Immun. 1976 Jul;14(1):129–134. doi: 10.1128/iai.14.1.129-134.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Rosen H., Klebanoff S. J. Chemiluminescence and superoxide production by myeloperoxidase-deficient leukocytes. J Clin Invest. 1976 Jul;58(1):50–60. doi: 10.1172/JCI108458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sagone A. L., Jr, King G. W., Metz E. N. A comparison of the metabolic response to phagocytosis in human granulocytes and monocytes. J Clin Invest. 1976 May;57(5):1352–1358. doi: 10.1172/JCI108403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Stanley P. E., Williams S. G. Use of the liquid scintillation spectrometer for determining adenosine triphosphate by the luciferase enzyme. Anal Biochem. 1969 Jun;29(3):381–392. doi: 10.1016/0003-2697(69)90323-6. [DOI] [PubMed] [Google Scholar]
  14. Stjernholm R. L., Allen R. C., Steele R. H., Waring W. W., Harris J. A. Impaired chemiluminescence during phagocytosis of opsonized bacteria. Infect Immun. 1973 Feb;7(2):313–314. doi: 10.1128/iai.7.2.313-314.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. WARDLAW A. C., PILLEMER L. The properdin system and immunity. V. The bactericidal activity of the properdin system. J Exp Med. 1956 May 1;103(5):553–575. doi: 10.1084/jem.103.5.553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Weening R. S., Wever R., Roos D. Quantitative aspects of the production of superoxide radicals by phagocytizing human granulocytes. J Lab Clin Med. 1975 Feb;85(2):245–252. [PubMed] [Google Scholar]

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