Neutrophil Chemotaxis Caused by Chronic Obstructive Pulmonary Disease Alveolar Macrophages: The Role of CXCL8 and the Receptors CXCR1/CXCR2

Alveolar macrophages produce neutrophil chemoattractants; this cellular cross-talk contributes to neutrophilic airway inflammation in chronic obstructive pulmonary disease (COPD). We have investigated the chemotaxis cross-talk mechanisms between these cells using COPD alveolar macrophages. Using conditioned media from stimulated COPD alveolar macrophages, we investigated the relative contributions of growth-related oncogene (CXCL1), interleukin-8 (CXCL8), and regulated on activation normal T cell expressed and secreted (CCL5) to neutrophil chemotaxis and evaluated the effect of blocking the chemokine receptors CXCR1 and CXCR2 on chemotaxis caused by macrophage-conditioned media. Furthermore, we evaluated whether corticosteroid treatment of stimulated alveolar macrophages inhibited the chemotaxis ability of conditioned media. Alveolar macrophages isolated from COPD (n = 8) and smoker (S) (n = 8) lungs were treated with ultra-pure lipopolysaccharide in the presence and absence of dexamethasone (1 μM). Supernatants were used for neutrophil chemotaxis assays. SB656933 (2-hydroxy-N,N-dimethyl-3-{2-[[(R)-1-(5-methyl-furan-2-yl)-propyl]amino]-3,4-dioxo-cyclobut-1-enylamino}-benzamide) (CXCR2 antagonist) and Sch527123 [1-(2-chloro-3-fluorophenyl)-3-(4-chloro-2-hydroxy-3-piperazin-1-ylsulfonylphenyl)urea, 3-(2-chloro-3-fluoro-phenyl)-1-(4-chloro-2-hydroxy-3-piperazin-1-ylsulfonyl-phenyl)urea] (dual CXCR1 and CXCR2 antagonist) and blocking antibodies for CXCL8, CXCL1, and CCL5 were assessed. Conditioned media caused neutrophil chemotaxis in COPD and smokers (60.5 and 79.9% of total cells, respectively). Dexamethasone did not significantly reduce neutrophil chemotaxis in COPD or S. SB656933 and Sch527123 inhibited chemotaxis in a concentration-dependent manner, with the dual antagonist Sch527123 causing greater inhibition of chemotaxis. CXCL8 antibody inhibited neutrophil chemotaxis to basal levels, although there was no significant effect of blocking either CXCL1 or CCL5 (P > 0.05). CXCL8 plays a major role in neutrophil chemotaxis caused by alveolar macrophage–derived conditioned media, and this is most effectively inhibited by dual antagonism of CXCR1 and CXCR2. Corticosteroids do not inhibit chemotaxis caused by macrophage-derived chemokines.

[1]  C. Harbron,et al.  Synergistic Effects of p38 Mitogen-Activated Protein Kinase Inhibition with a Corticosteroid in Alveolar Macrophages from Patients with Chronic Obstructive Pulmonary Disease , 2011, Journal of Pharmacology and Experimental Therapeutics.

[2]  D. Ray,et al.  Inhibition of Lipopolysaccharide-Stimulated Chronic Obstructive Pulmonary Disease Macrophage Inflammatory Gene Expression by Dexamethasone and the p38 Mitogen-Activated Protein Kinase Inhibitor N-cyano-N′-(2-{[8-(2,6-difluorophenyl)-4-(4-fluoro-2-methylphenyl)-7-oxo-7,8-dihydropyrido[2,3-d] pyrimidi , 2008, Journal of Pharmacology and Experimental Therapeutics.

[3]  P. Paré,et al.  The nature of small-airway obstruction in chronic obstructive pulmonary disease. , 2004, The New England journal of medicine.

[4]  Dave Singh,et al.  LPS challenge in healthy subjects: an investigation of neutrophil chemotaxis mechanisms involving CXCR1 and CXCR2. , 2012, International immunopharmacology.

[5]  O. Götze,et al.  The CXC-chemokine neutrophil-activating peptide-2 induces two distinct optima of neutrophil chemotaxis by differential interaction with interleukin-8 receptors CXCR-1 and CXCR-2. , 1997, Blood.

[6]  D. Ray,et al.  Reduced glucocorticoid receptor expression and function in airway neutrophils. , 2012, International immunopharmacology.

[7]  L. Fabbri,et al.  Severity of airflow limitation is associated with severity of airway inflammation in smokers. , 1998, American journal of respiratory and critical care medicine.

[8]  R. Louis,et al.  Leukotriene B4 contributes to exhaled breath condensate and sputum neutrophil chemotaxis in COPD. , 2009, Chest.

[9]  R. Tal-Singer,et al.  SB-656933, a novel CXCR2 selective antagonist, inhibits ex vivo neutrophil activation and ozone-induced airway inflammation in humans. , 2011, British journal of clinical pharmacology.

[10]  H. Magnussen,et al.  SCH527123, a novel CXCR2 antagonist, inhibits ozone-induced neutrophilia in healthy subjects , 2009, European Respiratory Journal.

[11]  R. Djukanović,et al.  Impaired neutrophil chemotaxis in chronic obstructive pulmonary disease. , 2007, American journal of respiratory and critical care medicine.

[12]  K. Horiuchi,et al.  Leukocyte ADAM17 Regulates Acute Pulmonary Inflammation , 2011, PloS one.

[13]  S. Farrow,et al.  IFN‐γ synergistically enhances LPS signalling in alveolar macrophages from COPD patients and controls by corticosteroid‐resistant STAT1 activation , 2012, British journal of pharmacology.

[14]  T. Yoneda,et al.  Airway inflammation in COPD assessed by sputum levels of interleukin-8. , 1997, Chest.

[15]  P. Barnes.  Mediators of Chronic Obstructive Pulmonary Disease , 2004, Pharmacological Reviews.

[16]  P. Barnes,et al.  Increased levels of the chemokines GROα and MCP-1 in sputum samples from patients with COPD , 2002, Thorax.

[17]  F. Bazzoni,et al.  The neutrophil as a cellular source of chemokines , 2000, Immunological reviews.

[18]  Colin McCann,et al.  LTB4 is a signal-relay molecule during neutrophil chemotaxis. , 2012, Developmental cell.

[19]  L. Palmberg,et al.  Increased neutrophil migration in smokers with or without chronic obstructive pulmonary disease , 2012, Respirology.

[20]  W. Falk,et al.  Rapid quantitation of neutrophil chemotaxis: use of a polyvinylpyrrolidone-free polycarbonate membrane in a multiwell assembly. , 1980, Journal of immunological methods.

[21]  R. Sakalauskas,et al.  The effect of induced sputum and bronchoalveolar lavage fluid from patients with chronic obstructive pulmonary disease on neutrophil migration in vitro. , 2010, Medicina.

[22]  K. Ley,et al.  DARC on RBC limits lung injury by balancing compartmental distribution of CXC chemokines , 2009, European journal of immunology.

[23]  C. Mackay,et al.  Different functions for the interleukin 8 receptors (IL-8R) of human neutrophil leukocytes: NADPH oxidase and phospholipase D are activated through IL-8R1 but not IL-8R2. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[24]  P. Barnes,et al.  Neutrophil chemotactic activity of sputum from patients with COPD: role of interleukin 8 and leukotriene B4. , 2003, Chest.

[25]  P. Jorens,et al.  Interleukin-8: an important chemoattractant in sputum of patients with chronic inflammatory airway diseases. , 1993, The American journal of physiology.

[26]  I. Pavord,et al.  Sputum eosinophilia and the short term response to inhaled mometasone in chronic obstructive pulmonary disease , 2005, Thorax.

[27]  T. Standiford,et al.  Regulation of human alveolar macrophage- and blood monocyte-derived interleukin-8 by prostaglandin E2 and dexamethasone. , 1992, American journal of respiratory cell and molecular biology.

[28]  P. Hiemstra,et al.  Monocyte chemoattractant protein 1 , interleukin 8 , and chronic airways in ̄ ammation in COPD , 2000 .

[29]  R. Stockley,et al.  The chemotactic activity of sputum from patients with bronchiectasis. , 1998, American journal of respiratory and critical care medicine.

[30]  J. Armstrong,et al.  Glucocorticoid sensitivity of lipopolysaccharide‐stimulated chronic obstructive pulmonary disease alveolar macrophages , 2009, Clinical and experimental immunology.

[31]  A. Gaggar,et al.  A Critical Role for LTA4H in Limiting Chronic Pulmonary Neutrophilic Inflammation , 2010, Science.

[32]  A. Gillissen,et al.  Inhibition of granulocyte migration by tiotropium bromide , 2011, Respiratory research.

[33]  N. Mukaida.  Pathophysiological roles of interleukin-8/CXCL8 in pulmonary diseases. , 2003, American journal of physiology. Lung cellular and molecular physiology.