Neutrophils (PMNs) from patients with adult respiratory distress syndrome (ARDS) were assessed for light scattering, membrane potential, and phagocytic responses using fluorescent probes and flow cytometry to evaluate individual cells. Qualitative and quantitative oxidant responses were measured by nitroblue tetrazolium (NBT) and cytochromec reduction assays, respectively. The results were correlated with the proportion of cells binding the PMN subset-specific monoclonal antibody 31D8. Despite an increased forward scatter signal (4.3±1.6 vs. 1.3±1.1 ARDS vs. control,P=0.041) and spontaneous NBT test (12.6±4.7% vs. 2.5 ±0.8% positive, ARDS vs. control,P=0.033) indicating in vivo priming of ARDS PMNs, there were no significant differences between ARDS and control PMNs in assays of stimulated membrane potential, NBT, and O·2 − production or phagocytosis. However, positive correlations between the degree of prestimulus forward light scatter and subsequent O·2 − production to FMLP (r=0.673,P=0.006) and between the percentage of bands and the O·2 − response to PMA (r=0.660,P =0.003), suggest that the great variability of the ARDS PMN functional responses may relate to varying degrees of in vivo cell priming and/or deactivation. ARDS PMNs demonstrated a significantly lower percentage of 31D8 positive cells (73.4 ±7.5% vs. 94.5±1.6%,P=0.012) and a lower level of 31D8 staining when compared to normals (60.1±10.4% of control level,P=0.001). The lower 31D8 expression did not directly correlate with any functional parameter tested or with the proportion of immature cells. However, patients receiving an intravenous PGE21-infusion demonstrated a significant increase in 31D8 staining relative to controls and inhibition of PMA-stimulated O·2 − production. The data suggest that the function of PMNs from ARDS patients varies widely and reflects great in vivo variation in cell priming. While the mechanism responsible for the lowered expression of the 31D8 antigen and its apparent modulation by PGE1 is unknown, 31D8 may be an indirect marker for in vivo stress factors that regulate the preferential release of a structurally distinct PMN subset from the bone marrow.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Tate, R. M., andJ. E. Repine. 1983. Neutrophils and the adult respiratory distress syndrome,State of the Art 97:552–559.
Fantone, J. C., andP. A. Ward. 1983. Mechanisms of neutrophil-dependent lung injury.In Handbook of Inflammation: Immunology of Inflammation. P. A. Ward, editor. Elsevier, New York.
Gallin, J. I. 1984. Human neutrophil heterogeneity exists but is it meaningful?Blood 63:977.
Seligmann, B. E., T. M. Chused, andJ. I. Gallin. 1982. Binding of fluoresceinated chemoattractant peptide to human neutrophils is heterogeneous and correlated with the heterogeneous stimulation of membrane potential changes.J. Cell. Biol. 95:1133.
Seligmann, B. E., D. A. Melnick, H. L. Malech, andJ. I. Gallin. 1985. An antibody binding to human neutrophils demonstrates antigenic heterogeneity detected early in myeloid maturation which correlates with functional heterogeneity of mature neutrophils.J. Immunol. 135:2647.
Brown, C. C., H. L. Malech, andJ. I. Gallin. 1987. Kinetics of 31D8 denned human neutrophil (N) subpopulations following intravenous endotoxin.Fed. Proc. 46(3):986.
Brown, C. C., H. L. Malech, C. F. Shrimpton, P. C. Beverly, T. Segal, andJ. I. Gallin. 1987. Unique human neutrophil populations defined by monoclonal antibody ED12F8C10.Clin. Res. 35:421A.
Fletcher, M. P., andB. E. Seligmann. 1986. PMN heterogeneity: Long-term stability of fluorescent membrane potential responses to the chemoattractantN-formyl-methionyl-leucylphenylalanine in healthy adults and correlation with respiratory burst activity.Blood 68:611.
Fletcher, M. P. 1986. Modulation of the heterogeneous membrane potential response of neutrophils toN-formyl-methionyl-leucyl-phenylalanine (FMLP) by leukotriene B4: Evidence for cell recruitment.J. Immunol. 136:4213–4219.
Fletcher, M. P., J. Gasson, andR. Weisbart. 1986. Human recombinant granulocyte-macrophage colony stimulating factor (GM-CSF) enhances the proportion of depolarizing neutrophils in response to the chemoattractant F-met-leu-phe (FMLP).Fed. Proc. 45:1103.
Seligmann, B. E., T. M. Chused, andJ. I. Gallin. 1984. Differential binding of chemoattractant peptide to subpopulations of human neutrophils.J. Immunol. 133:2641.
Seligmann, B. E., H. L. Malech, D. A. Melnick, andJ. I. Gallin. 1985. An antibody binding to human neutrophils demonstrates antigenic heterogeneity detected early in myeloid maturation which correlates with functional heterogeneity of mature neutrophils.J. Immunol. 135:2647.
Gallin, I. I., R. J. Jacobson, B. E. Seligmann, J. A. Metcalf, J. H. Mckay, R. A. Sacher, andH. L. Malech. 1986. A neutrophil membrane marker reveals two groups of chronic myelogenous leukemia and its absence may be a marker of disease progression.Blood 68:343–346.
Krause, P. J., H. L. Malech, J. Kristie, C. M. Kosciol, V. C. Herson, L. Eisenfeld, W. T. Pastuszak, A. Kraus, andB. Seligmann. 1986. Polymorphonuclear leukocyte heterogeneity in neonates and adults.Blood 68:200–204.
Holcroft, J. W., M. J. Vassar, andC. J. Weber. 1986. Prostaglandin E1 and survival in patients with the adult respiratory distress syndrome. A prospective trial.Ann. Surg. 203:371–378.
Fletcher, M. P. 1987. Prostaglandin E1 dissociates neutrophil (PMN) membrane potential and secretion responses to F-met-leu-phe (FMLP) from FMLP-induced shape change as assessed by flow cytometry.Clin. Res. 35:3.
Dunn, P. A., andH. W. Tyrer. 1981. Quantitation of neutrophil phagocytosis, using fluorescent latex beads.J. Lab. Clin. Med. 98:374.
Simchowitz, L., J. P. Atkinson, andI. Spilberg. 1980. Stimulus-specific deactivation of chemotactic factor-induced cyclic AMP response and Superoxide generation by human neutrophils.J. Clin. Invest. 66:736–747.
Donabedian, H., andJ. I. Gallin. 1981. Deactivation of human neutrophil chemotaxis by chemoattractants: Effect on receptors for the chemotactic factor f-met-leu-phe.J. Immunol. 127:839–844.
Fletcher, M. P., andB. E. Seligmann. 1985. Monitoring human neutrophil granule secretion by flow cytometry: Secretion and membrane potential changes assessed by light scatter and a fluorescent probe of membrane potential.J. Leuk. Biol. 37:431–447.
Zimmerman, G. A., A. D. Renzetti, andH. R. Hill. 1983. Functional and metabolic activity of granulocytes from patients with adult respiratory distress syndrome. Evidence for activated neutrophils in the pulmonary circulation.Am. Rev. Respir. Dis. 127:290–300.
Fantone, J. C., andD. A. Kinnes. 1983. Prostaglandin E1 and prostaglandin I2 modulation of Superoxide production by human neutrophils.Biochem. Biophys. Res. Commun. 113:506–512.
Sedgwick, J. B., M. L. Berube, andR. B. Zurier. 1985. Stimulus-dependent inhibition of Superoxide generation by prostaglandins.Clin. Immmol. Immunopathol. 34:205–215.
Fantone, J. C., S. L. Kunkel, andP. A. Ward. 1981. Suppression of human polymorphonuclear function after intravenous infusion of prostaglandin E1.Prostaglandins Med. 7:195–198.
Cox, J. W., N. A. Andreadis, R. C. Bone, R. J. Maunder, R. H. Pullen, J. J. Ursprung, andM. J. Vassar. 1988. Pulmonary extraction and pharmacokinetics of prostaglandin E1 during continuous intravenous infusion in patients with adult respiratory distress syndromeAm. Rev. Resp. Dis. 137:5–12.
This work was supported in part by NIH grant P30-DK35747, a University of California, Davis, Dean's Research Grant, and The Upjohn Company.
About this article
Cite this article
Fletcher, M.P., Vassar, M.J. & Holcroft, J.W. Patients with adult respiratory distress syndrome (ARDS) demonstrate in vivo neutrophil activation associated with diminished binding of neutrophil-specific monoclonal antibody 31D8. Inflammation 12, 455–473 (1988). https://doi.org/10.1007/BF00919439
- Nitroblue Tetrazolium
- Adult Respiratory Distress Syndrome
- Neutrophil Activation
- Forward Scatter