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Oxygen-derived free radicals producing activity and survival of activated polymorphonuclear leukocytes

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Activation of polymorphonuclear (PMN) leukocytes is known to generate oxygen free radicals (OFR). However the fate of activated PMN leukocytes is not known. We investigated the OFR producing (chemiluminescence) activity and the survival of the activated PMN leukocytes. The study was divided into two groups. Group I, In vivo study (n = 7): zymosan (8.4 mg/kg) was administered intravenously in the anesthetized dogs and the blood samples were collected before and after 5, 15, 30, 60 and 120 min of zymosan administration. This group represents the in vivo pre-stimulated PMN leukocytes; Group II, In vitro study (n = 7): the blood were collected from dogs and further divided into two groups. Group A (n = 7): non-stimulated, without any added zymosan and group B (n = 7): zymosan was added to stimulate PMN leukocytes. Blood samples from group A and B were also collected at various time intervals similar to in vivo studies. Oxygen free radical producing activity of PMN leukocytes was monitored by measuring luminoldependent chemiluminescence (CL). Opsonized zymosan was used to activate PMN leukocytes. The studies in which the PMN leukocytes were stimulated in in vivo, both oxygen derived free radicals and superoxide dismutase (SOD) inhibitable oxygen free radical CL decreased significantly for 60 min and tended to reach thereafter to the pre-stimulated values. The resting chemiluminescence (chemiluminescence without zymosan stimulation in the assay medium) increased significantly for 15 min reaching to pre-stimulated values at 30 min and thereafter. In in vitro studies, oxygen derived free radicals CL of pre-stimulated PMN leukocytes (Group B) was depressed for the whole duration of investigation while SOD inhibitable CL was depressed for only 60 min. There was approximately a two-fold increase in the resting CL within 5 min of PMN leukocyte activation and it remained high for the whole duration of study. The chemiluminescence of non-stimulated PMN leukocytes in vitro (group A) remained practically normal throughout the period of observation. In in vivo studies, total white blood cells (WBC) and PMN leukocyte counts decreased initially and tended to approach towards pre-stimulated values at the end of the protocol. There were no changes in these counts in in vitro studies. These results indicate that the capacity to generate OFR is decreased in the in vivo and in vitro pre-stimulated PMN leukocytes. However this activity recovers with time. This study also suggests that the activated PMN leukocytes are not destroyed.

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References

  1. Del Maestro RF, Thaw HH, Bjork J, Planker M, Arfors KE: Free radicals as mediators of tissue injury. Acta Physiol Scand 492 (Suppl): 43–57, 1980

    Google Scholar 

  2. Jolly SR, Kane WJ, Bailie MB, Abrams GD, Lucchesi BR: Canine myocardial reperfusion injury: Its reduction by the combined administration of superoxide dismutase and catalase. Circ Res 54: 277–285, 1984

    Google Scholar 

  3. Perkowski SZ, Havill AN, Flynn JT, Gee MH: Role of intrapulmonary release of eicosanoids and superoxide anion as mediators of pulmonary dysfunction of endothelial injury in sheep with intermittent complement activation. Circ Res 53: 574–583, 1983

    Google Scholar 

  4. Prasad K, Kalra J: Oxygen free radicals and heart failure. Angiology 39: 417–420, 1988

    Google Scholar 

  5. Prasad K, Kalra J, Buchko G. Acute hemorrhage and oxygen free radicals. Angiology 39: 1005–1013, 1988

    Google Scholar 

  6. Prasad K, Kalra J & Bharradwaj B: Increased chemiluminescence of polymorphonuclear leukocytes in dogs with volume overload heart failure. Br J Exp Path 70: 463–468, 1989

    Google Scholar 

  7. Prasad K, Kalra J, Chan WP, Chaudhary AK: Effect of oxygen free radicals on cardiovascular function at organ and cellular level. Am Heart J 117: 1196–1202, 1989

    Google Scholar 

  8. Hess ML, Okabe E, Kontos HA: Proton and free oxygen radical interaction with the calcium transport system of cardiac sarcoplasmic reticulum. J Mol Cell Cardiol 13: 767–772, 1981

    Google Scholar 

  9. Fantone JC, Ward PA: Role of oxygen-derived free radicals and metabolites in leukocytes-dependent inflammatory reactions. Am J Pathol 107: 397–417, 1982

    Google Scholar 

  10. Dzau VJ, Packer M, Lilly LS, Swartz SL, Hollenberg NK, Williams GH: Prostaglandins in severe congestive heart failure. Relation to activation of the Renin-Angiotensin system and hyponatremia. N Engl J Med 310: 347–352, 1984

    Google Scholar 

  11. Babior BM: The respiratory burst of phagocytes. J Clin Invest 73: 599–601, 1984

    Google Scholar 

  12. Kato T, Wokalek H, Schöpf E, Eggert H, Ernst M, Rietschel ET, Fischer H: Measurement of chemiluminescence in freshly drawn human blood. Klin Wochenschr 59: 203–211, 1981

    Google Scholar 

  13. LKB-Wallac 1251 Luminometer Manual. The study of phagocytosis and opsonisation using luminol enhanced chemiluminescence. Application Note 513. Wallac of Finland. pp 1–5, 1985

  14. Tono-oko T, Veno N, Matsumoto T: Chemiluminescence of whole blood. A simple and rapid method for the estimation to phagocytic function of granulocytes and opsonic activity in whole blood. Clin Immun Immunopath 26: 66–75, 1983

    Google Scholar 

  15. Hastings MJG, Petricevic I, Williams AJ, Cole PJ, Easmon CSF: The effect of culture media on the production and measurement of luminol-dependent chemiluminescence. Brit J Exp Path 63: 147–153, 1982

    Google Scholar 

  16. Holt ME, Ryall MET, Campbell AK: Albumin inhibits human polymorphonuclear leukocyte luminol-dependent chemiluminescence: evidence for oxygen free radical scavenging. Brit J Exp Path 65: 231–241, 1984

    Google Scholar 

  17. Nelson RD, Herron MJ, Schmidtke JR, Simmons RL: Chemiluminescence response of human leukocytes: Influence of medium components on light production. Infection Immun 17: 513–520, 1977

    Google Scholar 

  18. Dechatelet CR, Shirley PS: Evaluation of chronic granulomatous diseases by a chemiluminescent assay of microlitre quantities of whole blood. Clin Chem 27: 1739–1741, 1981

    Google Scholar 

  19. Dewald B, Baggioleni M, Curnette JT, Babior BM: Subcellular localization of the superoxide forming system in human neutrophils. J Clin Invest 63: 21–29, 1979

    Google Scholar 

  20. Godin DV, Wohaieb SA, Garnett ME, Goumeniouk AD: Antioxidant enzyme alterations in experimental and clinical diabetes. Mol Cell Biochem 84: 223–231, 1988

    Google Scholar 

  21. Guarnieri C, Flamigni F, Caldarera CM: Role of oxygen in the cellular damage induced by reoxygenation of hypoxic heart. J Mol Cell Cardiol 12: 797–808, 1980

    Google Scholar 

  22. Abdalla DSP, Monteiro HP, Oliveira JAC, Bechara EJH: Activities of superoxide dismutase and glutathione peroxidase in schizophrenic and manic-depressive patients. Clin Chem 32: 805–807, 1986

    Google Scholar 

  23. L'abbé MR, Fischer PWF: An automated method for the determination of Cu, Zn-superoxide dismutase in plasma and erythrocytes using an ABA-200TM discrete analyzer. Clin Biochem 19: 175–178, 1986

    Google Scholar 

  24. Hammerschmidt DE, Stroncek DF, Bowers TK, LammiKeefe CJ, Kurth DM, Andrew Ozalins BA, Nicoloff DM, Lillehei RC, Craddock PR, Jacob HS: Complement activation and neutropenia occuring during cardiopulmonary bypass. J Thorac Cardiovasc Surg 81: 370–377, 1981

    Google Scholar 

  25. Fehr J, Jacob HS: In vitro granulocyte adherence and in vivo margination. Two associated complement dependent functions. J Exp Med 146: 641–652, 1977

    Google Scholar 

  26. Craddock PR, Fehr J, Brigham KL, Kronenberg RS, Jacob HS: Complement and leukocyte mediated pulmonary dysfunction in hemodialysis. N Eng J Med 296: 769–774, 1977

    Google Scholar 

  27. Craddock PR, Hammerschmidt DE, White JG, Dalmasso AP, Jacob HS: Complement (C50)-induced granulocyte aggregration in vitro. A possible mechanism of complement mediated leukostasis and leukopenia. J Clin Invest 60: 261–264, 1977

    Google Scholar 

  28. Hammerschmidt DE, Craddock PR, McCullough J, Kronenberg RS, Dalmasso AP, Jacob HS: Complement activation and pulmonary leukostasis during nylon fiber filtration leukopharesis. Blood 51: 721–730, 1972

    Google Scholar 

  29. Engler RL, Dahlgren MD, Peterson MA, Schmid-Schoenbein G: Role of leukocytes in the response to acute myocardial ischemia and reflow in dogs. Am J Physiol 251: H314-H323, 1986

    Google Scholar 

  30. Engler RL, Dahlgren MA, Dobbs A, Schmid-Schoenbein G: Accumulation of polymorphonuclear leukocytes during 3 hour experimental myocardial ischemia. Am J Physiol 251: H93-H100, 1986

    Google Scholar 

  31. Forman MB, Puett DW, Virmani R: Endothelial and myocardial injury during ischemia and reperfusion: Pathogenesis and therapeutic implications. J Am Coll Cardiol 13: 450–459, 1989

    Google Scholar 

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Prasad, K., Chaudhary, A.K. & Kalra, J. Oxygen-derived free radicals producing activity and survival of activated polymorphonuclear leukocytes. Mol Cell Biochem 103, 51–62 (1991). https://doi.org/10.1007/BF00229593

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