Increased neutrophil mediator release in patients with pulmonary hypertension – suppression by inhaled iloprost

Frank Rose; Katja Hattar; Sybille Gakisch; Friedrich Grimminger; Horst Olschewski; Werner Seeger; Annette Tschuschner; Ralph T. Schermuly; Norbert Weissmann; Jörg Hanze; Ulf Sibelius; Hossein A. Ghofrani

doi:10.1160/TH03-03-0173

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2003; 90(06): 1141-1149
DOI: 10.1160/TH03-03-0173

Wound Healing and Inflammation/Infection

Schattauer GmbH

Increased neutrophil mediator release in patients with pulmonary hypertension – suppression by inhaled iloprost

Frank Rose

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Katja Hattar

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Sybille Gakisch

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Friedrich Grimminger

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Horst Olschewski

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Werner Seeger

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Annette Tschuschner

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Ralph T. Schermuly

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Norbert Weissmann

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Jörg Hanze

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Ulf Sibelius

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

,

Hossein A. Ghofrani

¹Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany

› Author Affiliations

Abstract

Summary

Polymorphonuclear neutrophils (PMN) have been implicated in various vascular inflammatory processes. We isolated PMN from venous blood samples of 10 patients with severe primary pulmonary arterial hypertension (PPH), 7 patients with pulmonary hypertension secondary to chronic thromboembolism (CTEPH), and 12 healthy controls. When stimulated with the calcium-ionophore A23187, platelet activating factor (PAF) or the microbial agent n-formyl-Methionyl-Leucyl-Phenylalanine (fMLP), significantly increased release of elastase and super-oxide anion was noted in both groups with pulmonary hyper-tension. Moreover, the neutrophils of CTEPH patients responded with an enhanced liberation of leukotriene (LT) B₄ and 5-hydroxyeicosatetraenoic acid (5-HETE). Inhalation of aerosolized iloprost (5 µg) caused a rapid decline in pulmonary vascular resistance, in both PPH and CTEPH. This hemodynamic response was paralleled by a significant suppression of ionophore- and ligand-induced elastase and superoxide release, as well as LTB₄ and 5-HETE formation. The neutrophil inhibitory effect of the inhalation maneuver was fully reproduced by in vitro incubation of neutrophils with 1-10 pg/ml iloprost for 2 hours. This is the first study to demonstrate that circulating neutrophils from patients with PPH and CTEPH possess an enhanced readiness to respond with inflammatory mediator generation to different stimulatory agents ex-vivo, and that PMN respiratory burst, elastase secretion and leukotriene generation are promptly reduced by an iloprost inhalation maneuver. Neutrophils might participate in the inflammatory processes in pulmonary arterial hypertension.

Keywords

Inflammation - prostaglandins - leukocytes - lung embolism - remodeling

Full Text

References

References
1 Zhang P, Summer WR, Bagby GJ. et al. Innate immunity and pulmonary host defense. Immunol Rev 2000; 173: 39-51.
2 Cirillo P, Golino P, Ragni M. et al. Activated platelets and leucocytes cooperatively stimulate smooth muscle cell proliferation and proto-oncogene expression via release of soluble growth factors. Cardiovasc Res 1999; 43: 210-8.
3 Robbins IM, Barst RJ Rubin LJ. et al. Increased levels of prostaglandin d(2) suggest macrophage activation in patients with primary pulmonary hypertension. Chest 2001; 120: 1639-44.
4 Brennan LA, Steinhorn RH, Wedgwood S. et al. Increased superoxide generation is associated with pulmonary hypertension in fetal lambs. A role for NADPH oxidase. Circ Res 2003; 92 (06) 683-91.
5 Lai YL, Wu HD, Chen CF. Antioxidants attenuate chronic hypoxic pulmonary hypertension. J Cardiovasc Pharmacol 1998; 32: 714-20.
6 Schwartz JD, Monea S, Marcus SG. et al. Soluble factor(s) released from neutrophils activates endothelial cell matrix metalloproteinase-2. J Surg Res 1998; 76: 79-85.
7 Skold CM, Liu X, Umino T. et al. Human neutrophil elastase augments fibroblast-mediated contraction of released collagen gels. Am J Respir Crit Care Med 1999; 159: 1138-46.
8 Zaidi SH, You XM, Ciura S. et al. Suppressed smooth muscle proliferation and inflammatory cell invasion after arterial injury in elafinoverexpressing mice. J Clin Invest 2000; 105: 1687-9.
9 Cowan KN, Heilbut A, Humpl T. et al. Complete reversal of fatal pulmonary hypertension in rats by a serine elastase inhibitor. Nat Med 2000; 6: 698-702.
10 Rabinovitch M. Elastase and the pathobiology of unexplained pulmonary hypertension. Chest 1998; 114: 213S-24S.
11 Wolters PJ, Chapman HA. Importance of lysosomal cysteine proteases in lung disease. Respir Res 2000; 1 (03) 170-7.
12 Olschewski H, Ghofrani HA, Walmrath D. Inhaled prostacyclin and iloprost in severe pulmonary hypertension secondary to lung fibrosis. Am J Respir Crit Care Med 1999; 160: 600-7.
13 Hoeper M, Schwarze M, Ehlerding S. et al. Long-term treatment of primary pulmonary hypertension with aerosolized iloprost, a prostacyclin analogue. N Engl J Med 2000; 342: 1866-70.
14 Olschewski H, Simonneau G, Galie N. et al. Aerosolized Iloprost Randomized Study Group. Inhaled iloprost for severe pulmonary hypertension. N Engl J Med 2002; 347 (05) 322-9.
15 Walmrath D, Schneider T, Schermuly R. et al. Direct comparison of inhaled nitric oxide and aerosolized prostacyclin in acute respiratory distress syndrome. Am J Respir Crit Care Med 1996; 153 (03) 991-6.
16 Mazzone A, Faggioli P, Cusa C. et al. Effects of iloprost on adhesion molecules and F1 + 2 in peripheral ischemia. Eur J Clin Invest 2002; 32 (12) 882-8.
17 Ghofrani HA, Rose F, Schermuly RT. et al. Amplification of the pulmonary vasodilatory response to inhaled iloprost by subthreshold phosphodiesterase types 3 and 4 inhibition in severe pulmonary hypertension. Crit Care Med 2002; 30 (11) 2489-92.
18 Korn RL, Fisher CA, Stenach N. et al. Iloprost reduces procoagulant activity in the extra-corporeal circuit. J Surg Res 1993; 55 (04) 433-40.
19 Beghetti M, Reber G, de MP. et al. Aerosolized iloprost induces a mild but sustained inhibition of platelet aggregation. Eur Respir J 2002; 19 (03) 518-24.
20 Riva CM, Morganroth ML, Ljungman AG. Iloprost inhibits neutrophil-induced lung injury and neutrophil adherence to endothelial monolayers. Am J Respir Cell Mol Biol 1990; 3: 301-9.
21 Mazzone A, Mazzucchelli I, Fossati G. Iloprost effects on phagocytes in patients suffering from ischaemic diseases: in vivo evidence for down-regulation of alpha M beta 2 integrin. Eur J Clin Invest 1996; 26: 860-6.
22 Grimminger F, Hattar K, Papavassilis C. et al. Neutrophil activation by anti-proteinase 3 antibodies in Wegener’s granulomatosis: role of exogenous arachidonic acid and leukotriene B4 generation. J Exp Med 1996; 184: 1567-72.
23 Rose F, Kiss L, Grimminger F. et al. E. coli hemolysin-induced lipid mediator metabolism in alveolar macrophages: impact of eicosapentaenoic acid. Am J Physiol Lung Cell Mol Physiol 2000; 279: L100-109.
24 Cohen HJ, Chovaniec ME. Superoxide generation by digitonin-stimulated guinea-pig granulocytes. J Clin Invest 1978; 61: 1081-7.
25 Kramps JA. L-Pyroglutamyl-L-propyl-L-valine-p-nitro-anilide, a highly specific substrate for granulocyte elastase. Scand J Clin Lab Invest 1983; 43: 427-32.
26 Khalfi F, Gressier B, Dine T. et al. Verapamil inhibits elastase release and superoxide anion production in human neutrophils. Biol Pharm Bull 1998; 21: 109-12.
27 Haga Y, Dumitrescu A, Zhang Y. et al. Effects of calcium blockers on the cytosolic calcium, H₂O₂ production and elastase release in human neutrophils. Pharmacol Toxicol 1996; 79: 312-7.
28 Boudier C, Cadene M, Bieth M. et al. Inhibiton of neutrophil cathepsin G by oxidizied mucus proteinase inhibitor. Effect of heparin. Biochemistry 1999; 38: 8451-7.
29 Bos JC, Grooteman MP, van Houte AJ. et al. Low polymorphonuclear cell degranulation during citrate anticoagulation: a comparison between citrate and heparin dialysis. Nephrol Dial Transplant 1997; 12: 1387-93.
30 Gessler T, Schmehl T, Hoeper M. et al. Ultrasonic versus jet nebulization of iloprost in severe pulmonary hypertension. Eur Respir J 2001; 17: 14-19.
31 Nilius SM, Hasse A, Kuger P. et al. Agonist-induced long-term desensitization of the human prostacyclin receptor. FEBS Lett 2000; 484 (03) 211-6.
32 Grimminger F, Kreusler B, Schneider U. et al. Influence of microvascular adherence on neutrophil leukotriene generation. Evidence for cooperative eicosanoid synthesis. J Immunol 1990; 144: 1866-72.