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
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DOI：
10.1124/jpet.112.201855
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发表时间：
2013-10-01
影响因子：
3.5
通讯作者：
Singh, Dave
中科院分区：
文献类型：
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作者：
Kaur, Manminder;Singh, Dave
肺泡巨噬细胞产生中性粒细胞趋化剂；这种细胞串扰会导致慢性阻塞性肺病（COPD）中的中性粒细胞气道炎症。我们使用慢性阻塞性肺病肺泡巨噬细胞研究了这些细胞之间的趋化串扰机制。使用来自受刺激的 COPD 肺泡巨噬细胞的条件培养基，我们研究了生长相关癌基因 (CXCL1)、白细胞介素 8 (CXCL8) 和调节正常 T 细胞表达和分泌的激活 (CCL5) 对中性粒细胞趋化性的相对贡献，并评估了阻断趋化因子受体 CXCR1 和 CXCR2 对巨噬细胞条件培养基引起的趋化性的影响。此外，我们评估了刺激的肺泡巨噬细胞的皮质类固醇治疗是否抑制条件培养基的趋化能力。在存在和不存在地塞米松 (1 μM) 的情况下，用超纯脂多糖处理从 COPD (n = 8) 和吸烟者 (S) (n = 8) 肺部分离的肺泡巨噬细胞。上清液用于中性粒细胞趋化性测定。 SB656933（2-羟基-N,N-二甲基-3-{2-[[(R)-1-(5-甲基-呋喃-2-基)-丙基]氨基]-3,4-二氧代-环丁-1-烯基氨基}-苯甲酰胺）（CXCR2拮抗剂）和Sch527123 [1-(2-氯-3-氟苯基)-3-(4-氯-2-羟基-3-哌嗪-1-基磺酰基苯基)脲、3-(2-氯-3-氟-苯基)-1-(4-氯-2-羟基-3-哌嗪-1-基磺酰基-苯基)脲]（CXCR1和CXCR2双重拮抗剂）和CXCL8、CXCL1和的封闭抗体对 CCL5 进行了评估。条件培养基引起 COPD 和吸烟者的中性粒细胞趋化性（分别占总细胞的 60.5% 和 79.9%）。地塞米松不会显着降低 COPD 或 S 中的中性粒细胞趋化性。SB656933 和 Sch527123 以浓度依赖性方式抑制趋化性，而双重拮抗剂 Sch527123 会引起更大的趋化性抑制。 CXCL8 抗体将中性粒细胞趋化性抑制至基础水平，但阻断 CXCL1 或 CCL5 没有显着效果 (P > 0.05)。 CXCL8 在肺泡巨噬细胞来源的条件培养基引起的中性粒细胞趋化性中起主要作用，CXCR1 和 CXCR2 的双重拮抗作用可最有效地抑制这一作用。皮质类固醇不会抑制巨噬细胞衍生的趋化因子引起的趋化作用。
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 mu M). 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.