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. 1980 Oct;66(4):736–747. doi: 10.1172/JCI109911

Stimulus-specific deactivation of chemotactic factor-induced cyclic AMP response and superoxide generation by human neutrophils.

L Simchowitz, J P Atkinson, I Spilberg
PMCID: PMC371648  PMID: 6252250

Abstract

The responses of isolated human peripheral neutrophils to either simultaneous or sequential additions of two chemotactic factors were studied. Simultaneous additions of formyl-methionyl-leucyl-phenylalanine (10-100 nM) and the fifth component of complement, C5a (1-10 microliters/ml), evoked partially additive responses of membrane depolarization as measured by the fluorescent dye 3,3'-dipropyl-thiocarbocyanine, a transient elevation of intracellular cyclic AMP (cAMP), and superoxide (O2-) generation as assessed by ferricytochrome c reduction. Preincubation of the cells with either formyl-methionyl-leucyl-phenylalanine or C5a alone caused dose-dependent inhibition of the depolarization, the cAMP increase, and O2- release induced by a subsequent exposure to an optimal dose of the same stimulus, i.e., deactivation occurred. In contrast, when cells were treated with one chemotactic factor and then exposed to the other stimulus, the cells exhibited a normal response of peak depolarization, the rise in cAMP, and O2-0 production i.e., cross-deactivation failed to occur. The results imply that deactivation of these phenomena is stimulus specific. Further, these observations are consistent with the hypothesis that cross-deactivation of chemotaxis is mediated by one or more processes that are irrelevant to O2- generation, and that occur distal to the depolarization and cAMP steps in the sequence of neutrophil activation: possibly microtubule polymerization and orientation.

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Selected References

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

  1. Atkinson J. P., Kelly J. P., Weiss A., Wedner H. J., Parker C. W. Enhanced intracellular cGMP concentrations and lectin-induced lymphocyte transformation. J Immunol. 1978 Dec;121(6):2282–2291. [PubMed] [Google Scholar]
  2. Atkinson J. P., Weiss A., Ito M., Kelly J., Parker C. W. Effects of ascorbic acid and sodium ascorbate on cyclic nucleotide metabolism in human lymphocytes. J Cyclic Nucleotide Res. 1979;5(2):107–123. [PubMed] [Google Scholar]
  3. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  4. Fantone J., Senior R. M., Kreutzer D. L., Jones M., Ward P. A. Biochemical quantitation of the chemotactic factor inactivator activity in human serum. J Lab Clin Med. 1979 Jan;93(1):17–24. [PubMed] [Google Scholar]
  5. Goetzl E. J., Austen K. F. Active site chemotactic factors and the regulation of the human neutrophil chemotactic response. Antibiot Chemother (1971) 1974;19:218–232. doi: 10.1159/000395433. [DOI] [PubMed] [Google Scholar]
  6. Harper J. F., Brooker G. Femtomole sensitive radioimmunoassay for cyclic AMP and cyclic GMP after 2'0 acetylation by acetic anhydride in aqueous solution. J Cyclic Nucleotide Res. 1975;1(4):207–218. [PubMed] [Google Scholar]
  7. Hatch G. E., Nichols W. K., Hill H. R. A simplified procedure for cyclic nucleotide radioimmunoassay and its application to human blood leukocytes. J Immunol Methods. 1978;19(4):301–308. doi: 10.1016/0022-1759(78)90014-5. [DOI] [PubMed] [Google Scholar]
  8. Hawkins D. Neutrophilic leukocytes in immunologic reactions in vitro. 3. Pharmacologic modulation of lysosomal constituent release. Clin Immunol Immunopathol. 1974 Jan;2(2):141–152. doi: 10.1016/0090-1229(74)90034-8. [DOI] [PubMed] [Google Scholar]
  9. Henson P. M. Pathologic mechanisms in neutrophil-mediated injury. Am J Pathol. 1972 Sep;68(3):593–612. [PMC free article] [PubMed] [Google Scholar]
  10. Henson P. M., Zanolari B., Schwartzman N. A., Hong S. R. Intracellular control of human neutrophil secretion. I. C5a-induced stimulus-specific desensitization and the effects of cytochalasin B. J Immunol. 1978 Sep;121(3):851–855. [PubMed] [Google Scholar]
  11. Jackowski S., Sha'afi R. I. Response of adenosine cyclic 3',5'-monophosphate level in rabbit neutrophils to the chemotactic peptide formyl-methionyl-leucyl-phenylalanine. Mol Pharmacol. 1979 Sep;16(2):473–481. [PubMed] [Google Scholar]
  12. Naccache P. H., Showell H. J., Becker E. L., Sha'afi R. I. Changes in ionic movements across rabbit polymorphonuclear leukocyte membranes during lysosomal enzyme release. Possible ionic basis for lysosomal enzyme release. J Cell Biol. 1977 Dec;75(3):635–649. doi: 10.1083/jcb.75.3.635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. O'Flaherty J. T., Kreutzer D. L., Showell H. J., Vitkauskas G., Becker E. L., Ward P. A. Selective neutrophil desensitization to chemotactic factors. J Cell Biol. 1979 Mar;80(3):564–572. doi: 10.1083/jcb.80.3.564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Showell H. J., Williams D., Becker E. L., Naccache P. H., Sha'afi R. Desensitization and deactivation of the secretory responsiveness of rabbit neutrophils induced by the chemotactic peptide, formyl-methionyl-leucyl-phenylalanine. J Reticuloendothel Soc. 1979 Feb;25(2):139–150. [PubMed] [Google Scholar]
  15. Simchowitz L., Fischbein L. C., Spilberg I., Atkinson J. P. Induction of a transient elevation in intracellular levels of adenosine-3',5'-cyclic monophosphate by chemotactic factors: an early event in human neutrophil activation. J Immunol. 1980 Mar;124(3):1482–1491. [PubMed] [Google Scholar]
  16. Simchowitz L., Mehta J., Spilberg I. Chemotactic factor-induced generation of superoxide radicals by human neutrophils: effect of metabolic inhibitors and antiinflammatory drugs. Arthritis Rheum. 1979 Jul;22(7):755–763. doi: 10.1002/art.1780220711. [DOI] [PubMed] [Google Scholar]
  17. Simchowitz L., Spilberg I. Chemotactic factor-induced generation of superoxide radicals by human neutrophils: evidence for the role of sodium. J Immunol. 1979 Nov;123(5):2428–2435. [PubMed] [Google Scholar]
  18. Simchowitz L., Spilberg I. Generation of superoxide radicals by human peripheral neutrophils activated by chemotactic factor. Evidence for the role of calcium. J Lab Clin Med. 1979 Apr;93(4):583–593. [PubMed] [Google Scholar]
  19. Sims P. J., Waggoner A. S., Wang C. H., Hoffman J. F. Studies on the mechanism by which cyanine dyes measure membrane potential in red blood cells and phosphatidylcholine vesicles. Biochemistry. 1974 Jul 30;13(16):3315–3330. doi: 10.1021/bi00713a022. [DOI] [PubMed] [Google Scholar]
  20. Smith C. W., Hollers J. C., Patrick R. A., Hassett C. Motility and adhesiveness in human neutrophils. Effects of chemotactic factors. J Clin Invest. 1979 Feb;63(2):221–229. doi: 10.1172/JCI109293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Spilberg I., Mandell B., Hoffstein S. A proposed model for chemotactic deactivation: evidence for microtubule modulation of polymorphonuclear leukocyte chemotaxis. J Lab Clin Med. 1979 Aug;94(2):361–369. [PubMed] [Google Scholar]
  22. Steiner A. L., Parker C. W., Kipnis D. M. Radioimmunoassay for cyclic nucleotides. I. Preparation of antibodies and iodinated cyclic nucleotides. J Biol Chem. 1972 Feb 25;247(4):1106–1113. [PubMed] [Google Scholar]
  23. Vitkauskas G., Showell H. J., Becker E. L. Specific binding of synthetic chemotactic peptides to rabbit peritoneal neutrophils: effects on dissociability of bound peptide, receptor activity and subsequent biologic responsiveness (deactivation). Mol Immunol. 1980 Feb;17(2):171–180. doi: 10.1016/0161-5890(80)90069-3. [DOI] [PubMed] [Google Scholar]
  24. Waggoner A. Optical probes of membrane potential. J Membr Biol. 1976 Jun 30;27(4):317–334. doi: 10.1007/BF01869143. [DOI] [PubMed] [Google Scholar]
  25. Ward P. A., Becker E. L. The deactivation of rabbit neutrophils by chemotactic factor and the nature of the activatable esterase. J Exp Med. 1968 Apr 1;127(4):693–709. doi: 10.1084/jem.127.4.693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Williams L. T., Snyderman R., Pike M. C., Lefkowitz R. J. Specific receptor sites for chemotactic peptides on human polymorphonuclear leukocytes. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1204–1208. doi: 10.1073/pnas.74.3.1204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. van GELDER B., SLATER E. C. The extinction coefficient of cytochrome c. Biochim Biophys Acta. 1962 Apr 23;58:593–595. doi: 10.1016/0006-3002(62)90073-2. [DOI] [PubMed] [Google Scholar]

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