Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1990 Nov 1;111(5):2171–2181. doi: 10.1083/jcb.111.5.2171

Tumor necrosis factor and CD11/CD18 (beta 2) integrins act synergistically to lower cAMP in human neutrophils

PMCID: PMC2116341  PMID: 1699953

Abstract

The ability of neutrophils (PMN) to undergo a prolonged respiratory burst in response to cytokines such as tumor necrosis factor-alpha (TNF) depends on expression of CD11/CD18 (beta 2) integrins and interaction with matrix protein-coated surfaces (Nathan, C., S. Srimal, C. Farber, E. Sanchez, L. Kabbash, A. Asch, J. Gailit, and S. D. Wright. 1989. J. Cell Biol. 109:1341-1349). We tested the hypothesis that changes in cAMP mediate the joint action of cytokines and integrins. When plated on FBS- or fibrinogen-coated surfaces, PMN responded to TNF with a sustained fall in intracellular cAMP. This did not occur without TNF; in suspended PMN; in PMN treated with anti-CD18 mAb; or in PMN genetically deficient in beta 2 integrins. A preceding fall in cAMP appeared essential for TNF to induce a respiratory burst, because drugs that elevate cAMP blocked the burst if added any time before, but not after, its onset. Adenosine analogues and cytochalasins also block the TNF-induced respiratory burst if added before, but not after, its onset. Both also blocked the TNF-induced fall in cAMP. The effect of cytochalasins led us to examine the relationship between cAMP and actin reorganization. The same conditions that led to a sustained fall in cAMP led at the same time to cell spreading and the assembly of actin filaments. As with the respiratory burst, cAMP-elevating agents inhibited TNF-induced cell spreading and actin filament assembly if added before, but not after, spreading began. Thus, occupation of TNF receptors and engagement of CD18 integrins interact synergistically in PMN to promote a fall in cAMP. The fall in cAMP is closely related to cell spreading and actin reorganization. These changes are necessary for TNF to induce a prolonged respiratory burst. We conclude that integrins can act jointly with cytokines to affect cell shape and function through alterations in the level of a second messenger, cAMP.

Full Text

The Full Text of this article is available as a PDF (2.4 MB).

Selected References

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

  1. Ambruso D. R., Bolscher B. G., Stokman P. M., Verhoeven A. J., Roos D. Assembly and activation of the NADPH:O2 oxidoreductase in human neutrophils after stimulation with phorbol myristate acetate. J Biol Chem. 1990 Jan 15;265(2):924–930. [PubMed] [Google Scholar]
  2. Becker E. L. The short and happy life of neutrophil activation. J Leukoc Biol. 1990 Apr;47(4):378–389. doi: 10.1002/jlb.47.4.378. [DOI] [PubMed] [Google Scholar]
  3. Berkow R. L., Wang D., Larrick J. W., Dodson R. W., Howard T. H. Enhancement of neutrophil superoxide production by preincubation with recombinant human tumor necrosis factor. J Immunol. 1987 Dec 1;139(11):3783–3791. [PubMed] [Google Scholar]
  4. Bourne H. R., Lehrer R. I., Cline M. J., Melmon K. L. Cyclic 3',5'-adenosine monophosphate in the human lukocyte: synthesis, degradation, andeffects n neutrophil candidacidal activity. J Clin Invest. 1971 Apr;50(4):920–929. doi: 10.1172/JCI106564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buyon J. P., Slade S. G., Reibman J., Abramson S. B., Philips M. R., Weissmann G., Winchester R. Constitutive and induced phosphorylation of the alpha- and beta-chains of the CD11/CD18 leukocyte integrin family. Relationship to adhesion-dependent functions. J Immunol. 1990 Jan 1;144(1):191–197. [PubMed] [Google Scholar]
  6. Chan D., Goate A., Puck T. T. Involvement of vimentin in the reverse transformation reaction. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2747–2751. doi: 10.1073/pnas.86.8.2747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chatila T. A., Geha R. S., Arnaout M. A. Constitutive and stimulus-induced phosphorylation of CD11/CD18 leukocyte adhesion molecules. J Cell Biol. 1989 Dec;109(6 Pt 2):3435–3444. doi: 10.1083/jcb.109.6.3435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coffey R. G., Davis J. S., Djeu J. Y. Stimulation of guanylate cyclase activity and reduction of adenylate cyclase activity by granulocyte-macrophage colony-stimulating factor in human blood neutrophils. J Immunol. 1988 Apr 15;140(8):2695–2701. [PubMed] [Google Scholar]
  9. Cox J. P., Karnovsky M. L. The depression of phagocytosis by exogenous cyclic nucleotides, prostaglandins, and theophylline. J Cell Biol. 1973 Nov;59(2 Pt 1):480–490. doi: 10.1083/jcb.59.2.480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cronstein B. N., Kramer S. B., Rosenstein E. D., Korchak H. M., Weissmann G., Hirschhorn R. Occupancy of adenosine receptors raises cyclic AMP alone and in synergy with occupancy of chemoattractant receptors and inhibits membrane depolarization. Biochem J. 1988 Jun 15;252(3):709–715. doi: 10.1042/bj2520709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cronstein B. N., Kramer S. B., Weissmann G., Hirschhorn R. Adenosine: a physiological modulator of superoxide anion generation by human neutrophils. J Exp Med. 1983 Oct 1;158(4):1160–1177. doi: 10.1084/jem.158.4.1160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Davies P., Fox R. I., Polyzonis M., Allison A. C., Haswell A. D. The inhibition of phagocytosis and facilitation of exocytosis in rabbit polymorphonuclear leukocytes by cytochalasin B. Lab Invest. 1973 Jan;28(1):16–22. [PubMed] [Google Scholar]
  13. De la Harpe J., Nathan C. F. A semi-automated micro-assay for H2O2 release by human blood monocytes and mouse peritoneal macrophages. J Immunol Methods. 1985 Apr 22;78(2):323–336. doi: 10.1016/0022-1759(85)90089-4. [DOI] [PubMed] [Google Scholar]
  14. Degerman E., Smith C. J., Tornqvist H., Vasta V., Belfrage P., Manganiello V. C. Evidence that insulin and isoprenaline activate the cGMP-inhibited low-Km cAMP phosphodiesterase in rat fat cells by phosphorylation. Proc Natl Acad Sci U S A. 1990 Jan;87(2):533–537. doi: 10.1073/pnas.87.2.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Detmers P. A., Wright S. D., Olsen E., Kimball B., Cohn Z. A. Aggregation of complement receptors on human neutrophils in the absence of ligand. J Cell Biol. 1987 Sep;105(3):1137–1145. doi: 10.1083/jcb.105.3.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dustin M. L., Springer T. A. T-cell receptor cross-linking transiently stimulates adhesiveness through LFA-1. Nature. 1989 Oct 19;341(6243):619–624. doi: 10.1038/341619a0. [DOI] [PubMed] [Google Scholar]
  17. Fällman M., Lew D. P., Stendahl O., Andersson T. Receptor-mediated phagocytosis in human neutrophils is associated with increased formation of inositol phosphates and diacylglycerol. Elevation in cytosolic free calcium and formation of inositol phosphates can be dissociated from accumulation of diacylglycerol. J Clin Invest. 1989 Sep;84(3):886–891. doi: 10.1172/JCI114249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gamble J. R., Harlan J. M., Klebanoff S. J., Vadas M. A. Stimulation of the adherence of neutrophils to umbilical vein endothelium by human recombinant tumor necrosis factor. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8667–8671. doi: 10.1073/pnas.82.24.8667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Heyworth C. M., Wallace A. V., Houslay M. D. Insulin and glucagon regulate the activation of two distinct membrane-bound cyclic AMP phosphodiesterases in hepatocytes. Biochem J. 1983 Jul 15;214(1):99–110. doi: 10.1042/bj2140099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hughes A. R., Martin M. W., Harden T. K. Pertussis toxin differentiates between two mechanisms of attenuation of cyclic AMP accumulation by muscarinic cholinergic receptors. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5680–5684. doi: 10.1073/pnas.81.18.5680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Iannone M. A., Wolberg G., Zimmerman T. P. Chemotactic peptide induces cAMP elevation in human neutrophils by amplification of the adenylate cyclase response to endogenously produced adenosine. J Biol Chem. 1989 Dec 5;264(34):20177–20180. [PubMed] [Google Scholar]
  22. Kramer I. M., van der Bend R. L., Verhoeven A. J., Roos D. The 47-kDa protein involved in the NADPH:O2 oxidoreductase activity of human neutrophils is phosphorylated by cyclic AMP-dependent protein kinase without induction of a respiratory burst. Biochim Biophys Acta. 1988 Sep 16;971(2):189–196. doi: 10.1016/0167-4889(88)90191-7. [DOI] [PubMed] [Google Scholar]
  23. Lamb N. J., Fernandez A., Conti M. A., Adelstein R., Glass D. B., Welch W. J., Feramisco J. R. Regulation of actin microfilament integrity in living nonmuscle cells by the cAMP-dependent protein kinase and the myosin light chain kinase. J Cell Biol. 1988 Jun;106(6):1955–1971. doi: 10.1083/jcb.106.6.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lamb N. J., Fernandez A., Feramisco J. R., Welch W. J. Modulation of vimentin containing intermediate filament distribution and phosphorylation in living fibroblasts by the cAMP-dependent protein kinase. J Cell Biol. 1989 Jun;108(6):2409–2422. doi: 10.1083/jcb.108.6.2409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Laudanna C., Miron S., Berton G., Rossi F. Tumor necrosis factor-alpha/cachectin activates the O2(-)-generating system of human neutrophils independently of the hydrolysis of phosphoinositides and the release of arachidonic acid. Biochem Biophys Res Commun. 1990 Jan 15;166(1):308–315. doi: 10.1016/0006-291x(90)91946-p. [DOI] [PubMed] [Google Scholar]
  26. Lehmeyer J. E., Johnston R. B., Jr Effect of anti-inflammatory drugs and agents that elevate intracellular cyclic AMP on the release of toxic oxygen metabolites by phagocytes: studies in a model of tissue-bound IgG. Clin Immunol Immunopathol. 1978 Apr;9(4):482–490. doi: 10.1016/0090-1229(78)90144-7. [DOI] [PubMed] [Google Scholar]
  27. Loetscher H., Pan Y. C., Lahm H. W., Gentz R., Brockhaus M., Tabuchi H., Lesslauer W. Molecular cloning and expression of the human 55 kd tumor necrosis factor receptor. Cell. 1990 Apr 20;61(2):351–359. doi: 10.1016/0092-8674(90)90815-v. [DOI] [PubMed] [Google Scholar]
  28. Marone G., Thomas L. L., Lichtenstein L. M. The role of agonists that activate adenylate cyclase in the control of cAMP metabolism and enzyme release by human polymorphonuclear leukocytes. J Immunol. 1980 Nov;125(5):2277–2283. [PubMed] [Google Scholar]
  29. Matsuyama T., Yamada A., Kay J., Yamada K. M., Akiyama S. K., Schlossman S. F., Morimoto C. Activation of CD4 cells by fibronectin and anti-CD3 antibody. A synergistic effect mediated by the VLA-5 fibronectin receptor complex. J Exp Med. 1989 Oct 1;170(4):1133–1148. doi: 10.1084/jem.170.4.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nathan C. F. Neutrophil activation on biological surfaces. Massive secretion of hydrogen peroxide in response to products of macrophages and lymphocytes. J Clin Invest. 1987 Dec;80(6):1550–1560. doi: 10.1172/JCI113241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nathan C. F. Respiratory burst in adherent human neutrophils: triggering by colony-stimulating factors CSF-GM and CSF-G. Blood. 1989 Jan;73(1):301–306. [PubMed] [Google Scholar]
  32. Nathan C., Srimal S., Farber C., Sanchez E., Kabbash L., Asch A., Gailit J., Wright S. D. Cytokine-induced respiratory burst of human neutrophils: dependence on extracellular matrix proteins and CD11/CD18 integrins. J Cell Biol. 1989 Sep;109(3):1341–1349. doi: 10.1083/jcb.109.3.1341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nourshargh S., Hoult J. R. Inhibition of human neutrophil degranulation by forskolin in the presence of phosphodiesterase inhibitors. Eur J Pharmacol. 1986 Mar 18;122(2):205–212. doi: 10.1016/0014-2999(86)90104-4. [DOI] [PubMed] [Google Scholar]
  34. RODRIGUEZ J., DEINHARDT F. Preparation of a semipermanent mounting medium for fluorescent antibody studies. Virology. 1960 Oct;12:316–317. doi: 10.1016/0042-6822(60)90205-1. [DOI] [PubMed] [Google Scholar]
  35. Richter J., Andersson T., Olsson I. Effect of tumor necrosis factor and granulocyte/macrophage colony-stimulating factor on neutrophil degranulation. J Immunol. 1989 May 1;142(9):3199–3205. [PubMed] [Google Scholar]
  36. Rivkin I., Rosenblatt J., Becker E. L. The role of cyclic AMP in the chemotactic responsiveness and spontaneous motility of rabbit peritoneal neutrophils. The inhibition of neutrophil movement and the elevation of cyclic AMP levels by catecholamines, prostaglandins, theophylline and cholera toxin. J Immunol. 1975 Oct;115(4):1126–1134. [PubMed] [Google Scholar]
  37. Sadler S. E., Maller J. L. In vivo regulation of cyclic AMP phosphodiesterase in Xenopus oocytes. Stimulation by insulin and insulin-like growth factor 1. J Biol Chem. 1987 Aug 5;262(22):10644–10650. [PubMed] [Google Scholar]
  38. Saltiel A. R. Insulin generates an enzyme modulator from hepatic plasma membranes: regulation of adenosine 3',5'-monophosphate phosphodiesterase, pyruvate dehydrogenase, and adenylate cyclase. Endocrinology. 1987 Mar;120(3):967–972. doi: 10.1210/endo-120-3-967. [DOI] [PubMed] [Google Scholar]
  39. Schall T. J., Lewis M., Koller K. J., Lee A., Rice G. C., Wong G. H., Gatanaga T., Granger G. A., Lentz R., Raab H. Molecular cloning and expression of a receptor for human tumor necrosis factor. Cell. 1990 Apr 20;61(2):361–370. doi: 10.1016/0092-8674(90)90816-w. [DOI] [PubMed] [Google Scholar]
  40. Schwartz M. A., Cragoe E. J., Jr, Lechene C. P. pH regulation in spread cells and round cells. J Biol Chem. 1990 Jan 25;265(3):1327–1332. [PubMed] [Google Scholar]
  41. Shappell S. B., Toman C., Anderson D. C., Taylor A. A., Entman M. L., Smith C. W. Mac-1 (CD11b/CD18) mediates adherence-dependent hydrogen peroxide production by human and canine neutrophils. J Immunol. 1990 Apr 1;144(7):2702–2711. [PubMed] [Google Scholar]
  42. Singer I. I., Scott S., Kawka D. W., Kazazis D. M. Adhesomes: specific granules containing receptors for laminin, C3bi/fibrinogen, fibronectin, and vitronectin in human polymorphonuclear leukocytes and monocytes. J Cell Biol. 1989 Dec;109(6 Pt 1):3169–3182. doi: 10.1083/jcb.109.6.3169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Smith C. A., Davis T., Anderson D., Solam L., Beckmann M. P., Jerzy R., Dower S. K., Cosman D., Goodwin R. G. A receptor for tumor necrosis factor defines an unusual family of cellular and viral proteins. Science. 1990 May 25;248(4958):1019–1023. doi: 10.1126/science.2160731. [DOI] [PubMed] [Google Scholar]
  44. Southwick F. S., Dabiri G. A., Paschetto M., Zigmond S. H. Polymorphonuclear leukocyte adherence induces actin polymerization by a transduction pathway which differs from that used by chemoattractants. J Cell Biol. 1989 Oct;109(4 Pt 1):1561–1569. doi: 10.1083/jcb.109.4.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Sporn S. A., Eierman D. F., Johnson C. E., Morris J., Martin G., Ladner M., Haskill S. Monocyte adherence results in selective induction of novel genes sharing homology with mediators of inflammation and tissue repair. J Immunol. 1990 Jun 1;144(11):4434–4441. [PubMed] [Google Scholar]
  46. Therrien S., Naccache P. H. Guanine nucleotide-induced polymerization of actin in electropermeabilized human neutrophils. J Cell Biol. 1989 Sep;109(3):1125–1132. doi: 10.1083/jcb.109.3.1125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tung P., Pai G., Johnson D. G., Punzalan R., Levin S. R. Relationships between adenylate cyclase and Na+, K(+)-ATPase in rat pancreatic islets. J Biol Chem. 1990 Mar 5;265(7):3936–3939. [PubMed] [Google Scholar]
  48. Turner C. E., Pavalko F. M., Burridge K. The role of phosphorylation and limited proteolytic cleavage of talin and vinculin in the disruption of focal adhesion integrity. J Biol Chem. 1989 Jul 15;264(20):11938–11944. [PubMed] [Google Scholar]
  49. Verghese M. W., Fox K., McPhail L. C., Snyderman R. Chemoattractant-elicited alterations of cAMP levels in human polymorphonuclear leukocytes require a Ca2+-dependent mechanism which is independent of transmembrane activation of adenylate cyclase. J Biol Chem. 1985 Jun 10;260(11):6769–6775. [PubMed] [Google Scholar]
  50. Wacholtz M. C., Patel S. S., Lipsky P. E. Leukocyte function-associated antigen 1 is an activation molecule for human T cells. J Exp Med. 1989 Aug 1;170(2):431–448. doi: 10.1084/jem.170.2.431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wright S. D., Meyer B. C. Phorbol esters cause sequential activation and deactivation of complement receptors on polymorphonuclear leukocytes. J Immunol. 1986 Mar 1;136(5):1759–1764. [PubMed] [Google Scholar]
  52. Wright S. D., Weitz J. I., Huang A. J., Levin S. M., Silverstein S. C., Loike J. D. Complement receptor type three (CD11b/CD18) of human polymorphonuclear leukocytes recognizes fibrinogen. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7734–7738. doi: 10.1073/pnas.85.20.7734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wymann M. P., Kernen P., Deranleau D. A., Baggiolini M. Respiratory burst oscillations in human neutrophils and their correlation with fluctuations in apparent cell shape. J Biol Chem. 1989 Sep 25;264(27):15829–15834. [PubMed] [Google Scholar]
  54. Yonehara S., Ishii A., Yonehara M. A cell-killing monoclonal antibody (anti-Fas) to a cell surface antigen co-downregulated with the receptor of tumor necrosis factor. J Exp Med. 1989 May 1;169(5):1747–1756. doi: 10.1084/jem.169.5.1747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Yuo A., Kitagawa S., Suzuki I., Urabe A., Okabe T., Saito M., Takaku F. Tumor necrosis factor as an activator of human granulocytes. Potentiation of the metabolisms triggered by the Ca2+-mobilizing agonists. J Immunol. 1989 Mar 1;142(5):1678–1684. [PubMed] [Google Scholar]
  56. Zhang Y. H., Lin J. X., Yip Y. K., Vilcek J. Enhancement of cAMP levels and of protein kinase activity by tumor necrosis factor and interleukin 1 in human fibroblasts: role in the induction of interleukin 6. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6802–6805. doi: 10.1073/pnas.85.18.6802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Zurier R. B., Hoffstein S., Weissmann G. Mechanisms of lysosomal enzyme release from human leukocytes. I. Effect of cyclic nucleotides and colchicine. J Cell Biol. 1973 Jul;58(1):27–41. doi: 10.1083/jcb.58.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. de la Harpe J., Nathan C. F. Adenosine regulates the respiratory burst of cytokine-triggered human neutrophils adherent to biologic surfaces. J Immunol. 1989 Jul 15;143(2):596–602. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES