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Exudate polymorphonuclear leukocytes isolated from skin chambers are primed for enhanced response to subsequent stimulation with chemoattractant f-Met-Leu-Phe and C3-opsonized yeast particles

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Abstract

The ability to respond metabolically to stimulation with both soluble and paniculate substances was investigated in human polymorphonuclear leukocytes (PMNLs) isolated from an aseptic inflammatory reaction. Exudate PMNLs isolated from skin chambers (E-PMNLs) and blood PMNLs isolated from the peripheral blood (B-PMNLs) of the same individual were investigated in parallel. E-PMNLs were primed, resulting in an increased chemiluminescence (CL) response to subsequent stimulation with the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (FMLP) (334%) and serum-opsonized yeast particles (C3 yeast) (201%), as compared to B-PMNLs. Phorbol myristate acetate (PMA) on the other hand, induced a CL response in E-PMNLs that was only 70% of the response obtained in B-PMNLs. A similar primed state resulting in enhancement of the CL response to FMLP and C3 yeast could be induced in B-PMNLs by pretreatment with a bacterial culture filtrate. Pretreatment of E-PMNLs with the bacterial culture filtrate, however, did not increase the CL response to FMLPs any further. The enhanced functional response to FMLPs in E-PMNLs was accompanied by an increased binding of the peptide, demonstrated by a doubling of the amount of bound f-Met-Leu-[3H]Phe (209%), as compared to B-PMNLs. The increased C3-yeast-induced CL generation in E-PMNLs was accompanied by an increased ingestion and attachment of C3-opsonized yeast particles. The enhancement of phagocytosis in E-PMNLs was, however, dependent upon the opsonin used, since IgG-opsonized yeast particles were phagocytosed to the same extent by E-PMNLs and B-PMNLs, thereby indicating that selective receptor modulation is also involved in the priming of E-PMNLs for an enhanced response to C3-yeast. These results show that exudate cells isolated from skin chambers are modulated with respect to receptor-mediated functions resulting in an increased metabolic response to FMLP coupled with an increased binding of the peptide and an increased phagocytosis of C3-coated yeast particles. Receptor modulation during exudation may be an important mechanism in regulating the inflammatory response by PMNLs.

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References

  1. Klebanoff, S. J., andR. A. Clark. 1978. The neutrophil: Function and Clinical Disorders. Elsevier/North-Holland Biomedical Press, Amsterdam.

    Google Scholar 

  2. Babior, B. M. 1984. The respiratory burst of phagocytes.J. Clin. Invest. 73:599.

    Google Scholar 

  3. Becker, E. L., H. J. Showell, P. M. Henson, andL. S. Hsu. 1974. The ability of chemotactic factors to induce lysosomal enzyme release. I. The characteristics of the release, the importance of surface and the relation of enzyme release to chemotactic responsiveness.J. Immunol. 112:2047.

    Google Scholar 

  4. Hed, J. 1981. Response of polymorphonuclear leukocytes to particles opsonized with IgG or C3b.Monogr. Allergy 17:92.

    Google Scholar 

  5. Wilkinson, P. C. 1982. Chemotaxis and Inflammation, 2nd ed. Churchill-Livingstone, Edinburgh.

    Google Scholar 

  6. Huey, R., andT. E. Hugli. 1985. Characterization of a C5a receptor on human polymorphonuclear leukocytes (PMN).J. Immunol. 135:2063.

    Google Scholar 

  7. Allen, R. A., A. J. Jesaitis, L. A. Sklar, C. G. Cochrane, andR. G. Painter. 1986. Physiochemical properties of theN-formyl peptide receptor on human neutrophils.J. Biol. Chem. 26:1854.

    Google Scholar 

  8. Goldman, D. W., andE. J. Goetzl. 1982. Specific binding of leukotriene B4 to receptors on human polymorphonuclear leukocytes.J. Immunol. 129:1600.

    Google Scholar 

  9. O'Flaherty, J. T., J. R. Surles, J. Redman, D. Jacobson, C. Piantadosi, andR. L. Wykle. 1986. Binding and metabolism of platelet-activating factor by human neutrophils.J. Clin. Invest. 78:381.

    Google Scholar 

  10. Berger, M., J. O'Shea, A. S. Cross, T. M. Folks, T. M. Chused, E. J. Brown, andM. M. Frank. 1984. Human neutrophils increase expression of C3bi as well as C3b receptors upon activation.J. Clin. Invest. 74:1566.

    Google Scholar 

  11. Fearon, D. T. 1980. Identification of the membrane glycoprotein that is the C3b receptor of the human erythrocyte, polymorphonuclear leukocyte, B lymphocyte, and monocyte.J. Exp. Med. 152:20.

    Google Scholar 

  12. Fleit, H. B., S. D. Wright, andJ. C. Unkeless. 1982. Human neutrophil Fc-gamma receptor distribution and structure.Proc. Natl. Acad. Sci. U.S.A. 79:3275.

    Google Scholar 

  13. Ginsburg, I., andP. Q. Quie. 1980. Modulation of human polymorphonuclear leukocyte chemotaxis by leukocyte extracts, bacterial products, inflammatory exudates, and polyelectrolytes.Inflammation 4:301.

    Google Scholar 

  14. Freischlag, J. B. Backstrom, D. Kelly, G. Keehn, andR. W. Busuttil. 1986. Comparison of blood and peritoneal neutrophil activity in rabbits with and without peritonitis.J. Surg. Res. 40:145.

    Google Scholar 

  15. Perianin, A., M. Roch-Arveller, J.-P. Giroud, andJ. Hakim. 1985. Effect of acute non-immune inflammation on locomotion of exudate and blood rabbit neutrophils.Inflammation 9:389.

    Google Scholar 

  16. Thomsen, P., L. M. Bjursten, andL. E. Ericson. 1986. Joint fluid leukocyte activation by preformed immune complexes.Inflammation 10:243.

    Google Scholar 

  17. Zimmerli, W., B. Seligmann, andJ. I. Gallin. 1986. Exudation primes human and guinea pig neutrophils for subsequent responsiveness to the chemotactic peptideN-formyl-methionyl-leucyl-phenylalanine and increases complement C3bi receptor expression.J. Clin. Invest. 77:925.

    Google Scholar 

  18. Fletcher, M. P., B. E. Seligmann, andJ. I. Gallin. 1982. Correlation of human neutrophil secretion, chemoattractant receptor mobilization, and enhanced functional capacity.J. Immunol. 128:941.

    Google Scholar 

  19. O'Shea, J. J., E. J. Brown, B. E. Seligmann, J. A. Metcalf, M. M. Frank, andJ. I. Gallin. 1985. Evidence for distinct intracellular pools of receptors for C3b and C3bi in human neutrophils.J. Immunol. 134:2580.

    Google Scholar 

  20. VanEpps, D. E., andM. L. Garcia. 1980. Enhancement of neutrophil function as a result of prior exposure to chemotactic factor.J. Clin. Invest. 66:167.

    Google Scholar 

  21. Briheim, G., O.Stendahl, and C.Dahlgren. 1987. The increased metabolic response by neutrophils isolated from patients with meningococcal meningitis can be simulated by an interaction in vitro between the phagocytes and meningococcal culture filtrate.J. Infect. Dis. (in press).

  22. Briheim, G., O.Stendahl, and C.Dahlgren. 1987. In vitro modulation of human neutrophil function by a culture filtrate fromNeisseria meningitidis (submitted).

  23. Hellum, K. B., andC. O. Solberg. 1977. Human leukocyte migration: Studies with an improved skin chamber technique.Acta Pathol. Microbiol. Scand. Sect. C. 85:413.

    Google Scholar 

  24. Scheja, A., andA. Forsgren. 1985. A skin chamber technique for leukocyte migration studies; description and reproducibility.Acta Pathol. Microbiol. Immunol. Scand. Sect. C. 93:25.

    Google Scholar 

  25. Boyum, A. 1968. Isolation of mononuclear cells and granulocytes from human blood. Isolation of mononuclear cell by one centrifugation and of granulocytes by combining centrifugation and sedimentation at 1g.Scand J. Clin. Lab. Invest. Suppl. 97:77.

    Google Scholar 

  26. Hed, J. 1977. The extinction of fluorescence by crystal violet and its use to differentiate between attached and ingested microorganisms in phagocytosis.FEMS Lett. 1:357.

    Google Scholar 

  27. Csato, M., A. Dobozy, L. Raith, andN. Simon. 1985. The function of polymorphonuclear leukocytes emigrating into the skin.Acta Derm.-Venereol. 65:106.

    Google Scholar 

  28. Wandall, J. H. 1982. Function of polymorphonuclear neutrophilic leucocytes. Comparison of leucocytes from blood and exudate in healthy volunteers.Acta Pathol. Microbiol. Immunol. Scand. Sect. C. 90:7.

    Google Scholar 

  29. Scheja, A., andA. Forsgren. 1985. Functional properties of polymorphonuclear leukocytes accumulated in a skin chamber.Acta Pathol. Microbiol. Immunol. Scand. Sect. C. 93:31.

    Google Scholar 

  30. Bentwood, B. J., andP. M. Henson. 1980. The sequential release of granule constituents from human neutrophils.J. Immunol. 124:855.

    Google Scholar 

  31. Nelson, R. D., R. T. McCormack, V. D. Fiegel, andR. L. Simmons. 1978. Chemotactic deactivation of human neutrophils: Evidence for nonspecific and specific components.Infect. Immun. 22:441.

    Google Scholar 

  32. Fletcher, M. P., andJ. I. Gallin. 1983. Human neutrophils contain an intracellular pool of putative receptors for the chemoattractantN-formyl-methionyl-leucyl-phenylalanine.Blood 62:792.

    Google Scholar 

  33. Gallin, J. I., M. P. Fletcher, B. E. Seligmann, S. Hoffstein, K. Cehrs, andN. Mounessa. 1982. Human neutrophil-specific granule deficiency: A model to assess the role of neutrophil-specific granules in the evolution of the inflammatory response.Blood 59:1317.

    Google Scholar 

  34. Gallin, J. I., J. A. Metcalf, D. Roos, B. E. Seligmann, andM. M. Friedman. 1984. Organelle-depleted human neutrophil cytoplasts used to study f-Met-Leu-Phe receptor modulation and cell function.J. Immunol. 133:415.

    Google Scholar 

  35. Gallin, J. I. 1984. Neutrophil specific granules: A fuse that ignites the inflammatory response.Clin. Res. 32:320.

    Google Scholar 

  36. Fletcher, M. P., andJ. I. Gallin. 1980. Degranulating stimuli increase the availability of receptors on human neutrophils for the chemoattractant f-Met-Leu-Phe.J. Immunol. 124:1585.

    Google Scholar 

  37. Fearon, D. T., andL. A. Collins. 1983. Increased expression of C3b receptors on polymorphonuclear leukocytes induced by chemotactic factors and by purification procedures.J. Immunol. 130:370.

    Google Scholar 

  38. Lee, J., R. M. Hakim, andD. T. Fearon. 1984. Increased expression of the C3b receptor by neutrophils and complement activation during haemodialysis.Clin. Exp. Immunol. 56:205.

    Google Scholar 

  39. Horan, T. D., D. English, andT. A. McPherson. 1982. Association of neutrophil chemiluminescence with microbicidal activity.Clin. Immun. Immunpathol. 22:259.

    Google Scholar 

  40. Sachs, T., C. F. Moldow, P. R. Craddock, T. K. Bowers, andH. S. Jacob. 1978. Oxygen radicals mediate endothelial cell damage by complement-stimulated granulocytes. An in vitro model of immune vascular damage.J. Clin. Invest. 61:1161.

    Google Scholar 

  41. Weiss, S. J., andA. F. LoBuglio. 1980. An oxygen-dependent mechanism of neutrophilmediated cytotoxicity.Blood 55:1020.

    Google Scholar 

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Briheim, G., Coble, B., Stendahl, O. et al. Exudate polymorphonuclear leukocytes isolated from skin chambers are primed for enhanced response to subsequent stimulation with chemoattractant f-Met-Leu-Phe and C3-opsonized yeast particles. Inflammation 12, 141–152 (1988). https://doi.org/10.1007/BF00916397

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