Summary
Luminol chemiluminescence induced by phagocytosis of bacteria was studied in a system consisting of polymorphnuclear granulocytes (PMN), serum, luminol and Staphylococcus aureus. To evaluate the quantitative relationships between luminol chemiluminescence and the bactericidal process time courses for both variables were compared. It was found that initial rate of increase of chemiluminescence and initial rate of killing of bacteria were well correlated whereas the correlation was poorer for later stages of the process. When the rate of the bactericidal process was varied by changing concentrations of bacteria and PMN, directly proportional variations of initial rates of increase of chemiluminescence were observed. This is interpreted as reflecting an accumulation of oxidizing radicals as the result of a phagocytosis dependent gradual activation of the NADPH oxidase system, leading to luminol oxidation and/or killing of bacteria. However, by thermal inactivation of PMNs, chemiluminescence could be diminished whereas killing remained essentially unaffected, showing that these two processes could be uncoupled. Also, addition of erythrocytes to the PMN suspension was associated with decreased chemiluminescence and lysis of erythrocytes with an increased chemiluminescence, emphazising the importance of proper control of the components of the leucocyte test suspension.
Similar content being viewed by others
References
Allen RC, Stjernholm RL, Steele RH (1972) Evidence for the generation of an electronic excitation state(s) in human polymorphonuclear leucocytes and its participation in bactericidal capacity. Biochem Biophys Res Commun 47: 679
Allen RC, Loose LD (1976) Phagocytic activation of a luminoldependent chemiluminescence in rabbit alveolar and peritoneal macrophages. Biochem Biophys Res Commun 69: 245
Babior BM (1978) Oxygen-dependent microbial killing by phagocytes. N Engl J Med 298: 659
Curnutte JT, Babior BM (1974) Biological defence mechanisms. The effect of bacteria and serum on superoxide production by granulocytes. J Clin Invest 53: 1662
Van Dyke KM, Trush M, Wilson M, Stealy P, Miles P (1977) Luminol-dependent chemiluminescence analysis of cellular and humoral defects of phagocytosis using a chemo-glo photometer. Microchem J 22: 463
Ewetz L, Thore A (1976) Factors affecting the specificity of the luminol reaction with hematin compounds. Anal Biochem 71: 564
Ewetz L, Palmblad J, Thore A (1979) Quantitation of phagocytosis by luminol chemiluminescence. In: Schram E, Stanley P (eds) Proceedings, International Symposium on Analytical Application of Bioluminescence and Chemiluminescence. State Printing Publ Inc, Westlake Village, CA, USA
Hodgson EK, Fridovich I (1973) The role of O381-1 in the chemiluminescence of luminol. Photochem Photobiol 18: 451
Lundin A, Thore A (1975) Analytical information obtainable by evaluation of the time course of firefly bioluminescence in the assay of ATP. Biochemistry 66: 47
Michelson AM (1973) Studies in bioluminescence. X. Chemical models of enzymic oxidations. Biochimie 55: 465
Nathan DG, Baehner RL (1971) Disorders of phagocytic cell function. Prog Hematol 7: 235
Palmblad J (1976) Fasting (acute energy deprivation) in man: Effect on polymorphonuclear granulocyte functions, plasma iron and serum transferrin, Scand J Haematol 17: 217
Palmblad J, Engstedt L (1979) Activation of the bactericidal capacity of blood granulocytes — Evaluation of a new method and the effects of levamisole. Acta Pathol Microbiol Scand [B] 23: 10
Solberg CO (1972) Evaluation of neutrophil granulocyte function. Acta Pathol Microbiol Scand [B] 80: 559
Stevens P, Winston DJ, Van Dyke K (1978) In vitro evaluation of opsonic and cellular granulocyte function by luminol-dependent chemiluminescence: Utility in patients with severe neutropenia and cellular deficiency States. Infect Immun 22: 41
Author information
Authors and Affiliations
Additional information
Supported by grants from the National Defence Research Institute
Rights and permissions
About this article
Cite this article
Ewetz, L., Palmblad, J. & Thore, A. The relationship between luminol chemiluminescence and killing of staphylococcus aureus by neutrophil granulocytes. Blut 43, 373–381 (1981). https://doi.org/10.1007/BF00320316
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00320316