Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1994 May 1;299(Pt 3):881–887. doi: 10.1042/bj2990881

Immunocomplexes stimulate different signalling events to chemoattractants in the neutrophil and regulate L-selectin and beta 2-integrin expression differently.

Y Molad 1, K A Haines 1, D C Anderson 1, J P Buyon 1, B N Cronstein 1
PMCID: PMC1138103  PMID: 7514872

Abstract

Neutrophils express receptors for numerous phlogistons which, when occupied, trigger distinct signal-transduction pathways. Previous studies have shown that stimulation of neutrophils with chemoattractants induces shedding of the adhesive molecule L-selectin and increased expression of the beta 2-integrin CD11b/CD18. We determined the effect of ligation of classic, G-protein-linked chemoattractant receptors [C5a, interleukin-8 (IL-8), formylmethionyl-leucylphenylalanine (FMLP) and substance P], receptors for the Fc portion of IgG (Fc gamma receptors) and receptors for transforming growth factor beta (TGF beta) on expression of adhesive molecules by neutrophils and the stimulus-transduction mechanisms thought to mediate these changes. We were surprised to observe that occupancy of Fc gamma receptors by immunocomplexes (BSA-anti-BSA) stimulated increased expression by neutrophils of CD11b/CD18 at concentrations which did not affect L-selectin expression (EC50 9 micrograms/ml versus 350 micrograms/ml respectively, P < 0.00001, n = 5). In contrast, similar to previous studies, recombinant C5a, recombinant IL-8 and FMLP all stimulated increased expression of CD11b/CD18 (170-260% of basal, P < 0.001, n = 5) and shedding of L-selectin (56-75% reduction from basal, P < 0.001, n = 5) at similar concentrations and with similar potencies (EC50 = 2, 5, and 3 nM respectively). In contrast, neither TGF beta 1 nor, surprisingly, substance P affected expression of CD11b/CD18 or L-selectin. The regulation of expression of CD11b/CD18 or L-selectin in response to FMLP or immunocomplexes was unaffected by cytochalasin B (5 micrograms/ml) or the tyrosine kinase inhibitor tyrphostin-25 (25 microM). Although occupancy of both chemoattractant (FMLP) and Fc gamma receptors stimulated increments in the second messenger diacylglycerol, disruption of actin microfilaments by cytochalasin B enhanced diacylglycerol generation in response to FMLP but not in response to ligation of Fc gamma receptors. Moreover, both FMLP and immune aggregates provoked fluxes of intracellular Ca2+ concentration which differed with respect to both magnitude and kinetics and did not correlate well with regulation of adhesive-molecule expression. As upregulation of CD11b/CD18 is tightly linked to exocytosis of specific granules, these results suggest that shedding of L-selectin by activated neutrophils is not linked to exocytosis. These studies provide further evidence that receptors for chemoattractants and immunocomplexes on the neutrophil are linked to multiple signalling pathways.

Full text

PDF
881

Selected References

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

  1. Altieri D. C., Edgington T. S. A monoclonal antibody reacting with distinct adhesion molecules defines a transition in the functional state of the receptor CD11b/CD18 (Mac-1). J Immunol. 1988 Oct 15;141(8):2656–2660. [PubMed] [Google Scholar]
  2. Anderson D. C., Miller L. J., Schmalstieg F. C., Rothlein R., Springer T. A. Contributions of the Mac-1 glycoprotein family to adherence-dependent granulocyte functions: structure-function assessments employing subunit-specific monoclonal antibodies. J Immunol. 1986 Jul 1;137(1):15–27. [PubMed] [Google Scholar]
  3. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  4. Berger M., O'Shea J., Cross A. S., Folks T. M., Chused T. M., Brown E. J., Frank M. M. Human neutrophils increase expression of C3bi as well as C3b receptors upon activation. J Clin Invest. 1984 Nov;74(5):1566–1571. doi: 10.1172/JCI111572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berkow R. L., Dodson R. W. Tyrosine-specific protein phosphorylation during activation of human neutrophils. Blood. 1990 Jun 15;75(12):2445–2452. [PubMed] [Google Scholar]
  6. Buyon J. P., Abramson S. B., Philips M. R., Slade S. G., Ross G. D., Weissmann G., Winchester R. J. Dissociation between increased surface expression of gp165/95 and homotypic neutrophil aggregation. J Immunol. 1988 May 1;140(9):3156–3160. [PubMed] [Google Scholar]
  7. Goldstein I. M., Roos D., Kaplan H. B., Weissmann G. Complement and immunoglobulins stimulate superoxide production by human leukocytes independently of phagocytosis. J Clin Invest. 1975 Nov;56(5):1155–1163. doi: 10.1172/JCI108191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goldstein I., Hoffstein S., Gallin J., Weissmann G. Mechanisms of lysosomal enzyme release from human leukocytes: microtubule assembly and membrane fusion induced by a component of complement. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2916–2920. doi: 10.1073/pnas.70.10.2916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gomez-Cambronero J., Huang C. K., Bonak V. A., Wang E., Casnellie J. E., Shiraishi T., Sha'afi R. I. Tyrosine phosphorylation in human neutrophil. Biochem Biophys Res Commun. 1989 Aug 15;162(3):1478–1485. doi: 10.1016/0006-291x(89)90841-3. [DOI] [PubMed] [Google Scholar]
  10. Griffin J. D., Spertini O., Ernst T. J., Belvin M. P., Levine H. B., Kanakura Y., Tedder T. F. Granulocyte-macrophage colony-stimulating factor and other cytokines regulate surface expression of the leukocyte adhesion molecule-1 on human neutrophils, monocytes, and their precursors. J Immunol. 1990 Jul 15;145(2):576–584. [PubMed] [Google Scholar]
  11. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  12. Haines K. A., Reibman J., Tang X. Y., Blake M., Weissmann G. Effects of protein I of Neisseria gonorrhoeae on neutrophil activation: generation of diacylglycerol from phosphatidylcholine via a specific phospholipase C is associated with exocytosis. J Cell Biol. 1991 Aug;114(3):433–442. doi: 10.1083/jcb.114.3.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Huang C. K., Bonak V., Laramee G. R., Casnellie J. E. Protein tyrosine phosphorylation in rabbit peritoneal neutrophils. Biochem J. 1990 Jul 15;269(2):431–436. doi: 10.1042/bj2690431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Huang C. K., Laramee G. R., Casnellie J. E. Chemotactic factor induced tyrosine phosphorylation of membrane associated proteins in rabbit peritoneal neutrophils. Biochem Biophys Res Commun. 1988 Mar 15;151(2):794–801. doi: 10.1016/s0006-291x(88)80351-6. [DOI] [PubMed] [Google Scholar]
  15. Jutila M. A., Kishimoto T. K., Finken M. Low-dose chymotrypsin treatment inhibits neutrophil migration into sites of inflammation in vivo: effects on Mac-1 and MEL-14 adhesion protein expression and function. Cell Immunol. 1991 Jan;132(1):201–214. doi: 10.1016/0008-8749(91)90019-8. [DOI] [PubMed] [Google Scholar]
  16. Kishimoto T. K., Jutila M. A., Berg E. L., Butcher E. C. Neutrophil Mac-1 and MEL-14 adhesion proteins inversely regulated by chemotactic factors. Science. 1989 Sep 15;245(4923):1238–1241. doi: 10.1126/science.2551036. [DOI] [PubMed] [Google Scholar]
  17. Kishimoto T. K., Jutila M. A., Butcher E. C. Identification of a human peripheral lymph node homing receptor: a rapidly down-regulated adhesion molecule. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2244–2248. doi: 10.1073/pnas.87.6.2244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kolasinski S. L., Haines K. A., Siegel E. L., Cronstein B. N., Abramson S. B. Neuropeptides and inflammation. A somatostatin analog as a selective antagonist of neutrophil activation by substance P. Arthritis Rheum. 1992 Apr;35(4):369–375. doi: 10.1002/art.1780350402. [DOI] [PubMed] [Google Scholar]
  19. Lefkowitz R. J. Thrombin receptor. Variations on a theme. Nature. 1991 May 30;351(6325):353–354. doi: 10.1038/351353a0. [DOI] [PubMed] [Google Scholar]
  20. Miller L. J., Bainton D. F., Borregaard N., Springer T. A. Stimulated mobilization of monocyte Mac-1 and p150,95 adhesion proteins from an intracellular vesicular compartment to the cell surface. J Clin Invest. 1987 Aug;80(2):535–544. doi: 10.1172/JCI113102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Naccache P. H., Gilbert C., Caon A. C., Gaudry M., Huang C. K., Bonak V. A., Umezawa K., McColl S. R. Selective inhibition of human neutrophil functional responsiveness by erbstatin, an inhibitor of tyrosine protein kinase. Blood. 1990 Nov 15;76(10):2098–2104. [PubMed] [Google Scholar]
  22. Nasmith P. E., Mills G. B., Grinstein S. Guanine nucleotides induce tyrosine phosphorylation and activation of the respiratory burst in neutrophils. Biochem J. 1989 Feb 1;257(3):893–897. doi: 10.1042/bj2570893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. O'Shea J. J., Brown E. J., Seligmann B. E., Metcalf J. A., Frank M. M., Gallin J. I. Evidence for distinct intracellular pools of receptors for C3b and C3bi in human neutrophils. J Immunol. 1985 Apr;134(4):2580–2587. [PubMed] [Google Scholar]
  24. Philips M. R., Buyon J. P., Winchester R., Weissmann G., Abramson S. B. Up-regulation of the iC3b receptor (CR3) is neither necessary nor sufficient to promote neutrophil aggregation. J Clin Invest. 1988 Aug;82(2):495–501. doi: 10.1172/JCI113623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Reibman J., Haines K. A., Gude D., Weissmann G. Differences in signal transduction between Fc gamma receptors (Fc gamma RII, Fc gamma RIII) and FMLP receptors in neutrophils. Effects of colchicine on pertussis toxin sensitivity and diacylglycerol formation. J Immunol. 1991 Feb 1;146(3):988–996. [PubMed] [Google Scholar]
  26. Reibman J., Korchak H. M., Vosshall L. B., Haines K. A., Rich A. M., Weissmann G. Changes in diacylglycerol labeling, cell shape, and protein phosphorylation distinguish "triggering" from "activation" of human neutrophils. J Biol Chem. 1988 May 5;263(13):6322–6328. [PubMed] [Google Scholar]
  27. Reibman J., Meixler S., Lee T. C., Gold L. I., Cronstein B. N., Haines K. A., Kolasinski S. L., Weissmann G. Transforming growth factor beta 1, a potent chemoattractant for human neutrophils, bypasses classic signal-transduction pathways. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6805–6809. doi: 10.1073/pnas.88.15.6805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Roos D., Homan-Müller J. W., Weening R. S. Effect of cytochalasin B on the oxidative metabolism of human peripheral blood granulocytes. Biochem Biophys Res Commun. 1976 Jan 12;68(1):43–50. doi: 10.1016/0006-291x(76)90007-3. [DOI] [PubMed] [Google Scholar]
  29. Schwartz B. R., Ochs H. D., Beatty P. G., Harlan J. M. A monoclonal antibody-defined membrane antigen complex is required for neutrophil-neutrophil aggregation. Blood. 1985 Jun;65(6):1553–1556. [PubMed] [Google Scholar]
  30. Serra M. C., Bazzoni F., Della Bianca V., Greskowiak M., Rossi F. Activation of human neutrophils by substance P. Effect on oxidative metabolism, exocytosis, cytosolic Ca2+ concentration and inositol phosphate formation. J Immunol. 1988 Sep 15;141(6):2118–2124. [PubMed] [Google Scholar]
  31. Smith C. W., Kishimoto T. K., Abbassi O., Hughes B., Rothlein R., McIntire L. V., Butcher E., Anderson D. C., Abbass O. Chemotactic factors regulate lectin adhesion molecule 1 (LECAM-1)-dependent neutrophil adhesion to cytokine-stimulated endothelial cells in vitro. J Clin Invest. 1991 Feb;87(2):609–618. doi: 10.1172/JCI115037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Smith C. W., Rothlein R., Hughes B. J., Mariscalco M. M., Rudloff H. E., Schmalstieg F. C., Anderson D. C. Recognition of an endothelial determinant for CD 18-dependent human neutrophil adherence and transendothelial migration. J Clin Invest. 1988 Nov;82(5):1746–1756. doi: 10.1172/JCI113788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Snyderman R., Smith C. D., Verghese M. W. Model for leukocyte regulation by chemoattractant receptors: roles of a guanine nucleotide regulatory protein and polyphosphoinositide metabolism. J Leukoc Biol. 1986 Dec;40(6):785–800. doi: 10.1002/jlb.40.6.785. [DOI] [PubMed] [Google Scholar]
  34. Stevenson K. B., Nauseef W. M., Clark R. A. The neutrophil glycoprotein Mo1 is an integral membrane protein of plasma membranes and specific granules. J Immunol. 1987 Dec 1;139(11):3759–3763. [PubMed] [Google Scholar]
  35. Todd R. F., 3rd, Arnaout M. A., Rosin R. E., Crowley C. A., Peters W. A., Babior B. M. Subcellular localization of the large subunit of Mo1 (Mo1 alpha; formerly gp 110), a surface glycoprotein associated with neutrophil adhesion. J Clin Invest. 1984 Oct;74(4):1280–1290. doi: 10.1172/JCI111538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vedder N. B., Harlan J. M. Increased surface expression of CD11b/CD18 (Mac-1) is not required for stimulated neutrophil adherence to cultured endothelium. J Clin Invest. 1988 Mar;81(3):676–682. doi: 10.1172/JCI113372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ward P. A., Zvaifler N. J. Quantitative phagocytosis by neutrophils. I. A new method with immune complexes. J Immunol. 1973 Dec;111(6):1771–1776. [PubMed] [Google Scholar]
  38. Weissmann G., Smolen J. E., Korchak H. M. Release of inflammatory mediators from stimulated neutrophils. N Engl J Med. 1980 Jul 3;303(1):27–34. doi: 10.1056/NEJM198007033030109. [DOI] [PubMed] [Google Scholar]
  39. Zigmond S. H., Hirsch J. G. Effects of cytochalasin B on polymorphonuclear leucocyte locomotion, phagocytosis and glycolysis. Exp Cell Res. 1972 Aug;73(2):383–393. doi: 10.1016/0014-4827(72)90062-6. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES