LPS challenge in healthy subjects: An investigation of neutrophil chemotaxis mechanisms involving CXCR1 and CXCR2

Aul, Raminder; Patel, Sheena; Summerhill, Susan; Kilty, Iain; Plumb, Jonathan; Singh, Dave

doi:10.1016/j.intimp.2012.04.008

Cited by 29 publications

(39 citation statements)

References 34 publications

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“…Representative images of n = 3 independent experiments are shown. SB656933 on neutrophil chemotaxis, highlighting the potential advantage of targeting CXCR1 in addition to CXCR2 (Aul et al, 2012). The current study similarly showed a greater effect of Sch527123, which caused greater inhibition than SB656933 at the highest concentration, and showed significant inhibitory effects at lower concentrations (i.e., 1 mM).…”

Section: Discussionsupporting

confidence: 66%

“…We previously compared these drugs using sputum supernatant obtained from healthy patients after inhaled LPS challenge; this model was used to simulate the mixture of cytokines present that cause neutrophilic lung disease (Aul et al, 2012). We observed a greater effect of Sch527123 compared with Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Induced sputum supernatants have been used to investigate neutrophil chemotaxis (Richman-Eisenstat et al, 1993;Mikami et al, 1998;Beeh et al, 2003;Babusyte et al, 2010;Aul et al, 2012); this method is physiologically relevant as a number of chemokines are present in the media. We have recently shown that induced sputum supernatants from healthy subjects, obtained after lipopolysaccharide (LPS) inhalation, cause a greater degree of chemotaxis compared with the same concentrations of individual chemokines found in the sputum (Aul et al, 2012). Furthermore, dual antagonists blocking CXCR1/2 had a greater inhibitory effect on sputum supernatant-induced neutrophil chemotaxis compared with CXCR2 blockade alone.…”

Section: Introductionmentioning

confidence: 99%

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

Kaur

Singh

2013

J Pharmacol Exp Ther

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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 growthrelated 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 5 8) and smoker (S) (n 5 8) lungs were treated with ultra-pure lipopolysaccharide in the presence and absence of dexamethasone (1 mM). 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 macrophagederived chemokines.

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Section: Discussionsupporting

confidence: 66%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

Kaur

Singh

2013

J Pharmacol Exp Ther

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“…The chemotaxis assay was performed using 96-well Multiscreen™ MIC plates incorporating 5 μm pore diameter polycarbonate filter membranes (Merck-Millipore, Watford-UK) based on the method of transwell diffusion [27,28]. CD14+ monocytes (1 × 10 5 cells/well) were applied to the upper plate and incubated with either anti-human CCR5 monoclonal antibody (mAb) (20 μg/ml, R&D Systems) or PBS alone.…”

Section: Methodsmentioning

confidence: 99%

“…Migrated cells in the lower plate were collected, incubated with lysis buffer containing Quant-IT PicoGreen fluorescent dsDNA binding probe (Life Technologies, Paisley-UK) and analysed using a BMG LabTech FluoStar Optima optical density plate reader. CD14+ monocyte chemotaxis was expressed as a proportion (%) of maximum CD14+ monocyte migration (CD14+ cells added directly to the wells of the lower plate) [28]. All conditions were assayed in triplicate.…”

Section: Methodsmentioning

confidence: 99%

Increased levels of soluble interleukin-6 receptor and CCL3 in COPD sputum

et al. 2014

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BackgroundCOPD patients have increased numbers of macrophages and neutrophils in the lungs. Interleukin-6 (IL-6) trans-signaling via its soluble receptor sIL-6R, governs the influx of innate immune cells to inflammatory foci through regulation of the chemokine CCL3. We hypothesized that there would be enhanced levels of IL-6, sIL-6R and CCL3 in COPD sputum.Methods59 COPD patients, 15 HNS and 15 S underwent sputum induction and processing with phosphate buffered saline to obtain supernatants for IL-6, sIL-6R and CCL3 analysis. Cytoslides were produced for differential cell counting and immunocytochemistry (COPD; n = 3) to determine cell type surface expression of the CCL3 receptors CCR5 and CCR1.ResultsCOPD patients expressed higher levels (p < 0.05) of sIL-6R and CCL3 compared to controls (sIL-6R medians pg/ml: COPD 166.4 vs S 101.1 vs HNS 96.4; CCL3 medians pg/ml: COPD 117.9 vs S 0 vs HNS 2.7). COPD sIL-6R levels were significantly correlated with sputum neutrophil (r = 0.5, p < 0.0001) and macrophage (r = 0.3, p = 0.01) counts. Immunocytochemical analysis revealed that CCR5 and CCR1 were exclusively expressed on airway macrophages.ConclusionEnhanced airway generation of sIL-6R may promote IL-6 trans-signaling in COPD. Associated upregulation of CCL3 may facilitate the recruitment of macrophages into the airways by ligation of CCR1 and CCR5.Electronic supplementary materialThe online version of this article (doi:10.1186/s12931-014-0103-4) contains supplementary material, which is available to authorized users.

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Boronic acid-containing aminopyridine- and aminopyrimidinecarboxamide CXCR1/2 antagonists: Optimization of aqueous solubility and oral bioavailability

Schuler

Engles

Maeda

et al. 2015

Bioorganic & Medicinal Chemistry Letters

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The chemokine receptors CXCR1 and CXCR2 are important pharmaceutical targets due to their key roles in inflammatory diseases and cancer progression. We have previously identified 2-[5-(4-Fluoro-phenylcarbamoyl)-pyridin-2-ylsulfanylmethyl]-phenylboronic acid (SX-517) and 6-(2-boronic acid-5-trifluoromethoxy-benzylsulfanyl)-N-(4-fluoro-phenyl)-nicotinamide (SX-576) as potent non-competitive boronic acid-containing CXCR1/2 antagonists. Herein we report the synthesis and evaluation of aminopyridine and aminopyrimidine analogues of SX-517 and SX-576, identifying (2-{(Benzyl)[(5-boronic acid-2-pyridyl)methyl]amino}-5-pyrimidinyl)(4-fluorophenylamino)formaldehyde as a potent chemokine antagonist with improved aqueous solubility and oral bioavailability.

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LPS challenge in healthy subjects: An investigation of neutrophil chemotaxis mechanisms involving CXCR1 and CXCR2

Cited by 29 publications

References 34 publications

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

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

Increased levels of soluble interleukin-6 receptor and CCL3 in COPD sputum

Boronic acid-containing aminopyridine- and aminopyrimidinecarboxamide CXCR1/2 antagonists: Optimization of aqueous solubility and oral bioavailability

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