Airway Disease, National Heart and Lung Institute, Imperial College London, London, United Kingdom.
Airway Disease, National Heart and Lung Institute, Imperial College London, London, United Kingdom; Department of Histopathology, Royal Brompton and Harefield NHS Foundation Trust, London, United Kingdom.
J Allergy Clin Immunol. 2014 Jan;133(1):207-16.e1-11. doi: 10.1016/j.jaci.2013.08.044. Epub 2013 Oct 28.
In patients with chronic obstructive pulmonary disease (COPD), pulmonary macrophages increase in number, release increased levels of inflammatory mediators, and respond poorly to glucocorticosteroids. Whether this is due to a change in macrophage phenotype or localized activation is unknown.
We sought to investigate whether macrophages from patients with COPD are a distinct phenotype.
Macrophage populations were isolated from human lung tissue from nonsmokers, smokers, and patients with COPD by using Percoll density gradients. Five macrophage populations were isolated on the basis of density (1.011-1.023, 1.023-1.036, 1.036-1.048, 1.048-1.061, and 1.061-1.073 g/mL), and cell-surface expression of CD14, CD16, CD163, CD40, and CD206 was assessed by using flow cytometry. Release of active matrix metalloproteinase 9, TNF-α, CXCL8, and IL-10 was measured by using ELISA.
The 2 least dense fractions were more than 90% apoptotic/necrotic, with the remaining fractions greater than 70% viable. Macrophages from nonsmokers and smokers were CD163(+), CD206(+), CD14(+), and CD40(-), whereas macrophages from patients with COPD were less defined, showing significantly lower expression of all receptors. There were no differences in receptor expression associated with density. Macrophages from patients with COPD of a density of 1.036 to 1.048 g/mL released higher levels of active matrix metalloproteinase 9 compared with cells from nonsmokers, with no difference between the remaining fractions. This population of macrophages from patients with COPD was less responsive to budesonide compared with those from nonsmokers and smokers when stimulated with LPS. Glucocorticosteroid insensitivity was selective for proinflammatory cytokines because budesonide inhibition of LPS-stimulated IL-10 release was similar for all macrophages.
This study identifies a specific macrophage phenotype in the lungs of patients with COPD who are glucocorticosteroid insensitive with a density of 1.036 to 1.048 g/mL but do not correspond to the current concept of macrophage phenotypes.
在慢性阻塞性肺疾病（COPD）患者中，肺巨噬细胞数量增加，释放的炎症介质水平升高，并且对糖皮质激素的反应不佳。这是否是由于巨噬细胞表型的改变或局部激活尚不清楚。
我们试图研究 COPD 患者的巨噬细胞是否是一种独特的表型。
通过 Percoll 密度梯度从非吸烟者、吸烟者和 COPD 患者的肺组织中分离巨噬细胞群体。根据密度（1.011-1.023、1.023-1.036、1.036-1.048、1.048-1.061 和 1.061-1.073 g/mL）分离了 5 种巨噬细胞群体，并通过流式细胞术评估细胞表面 CD14、CD16、CD163、CD40 和 CD206 的表达。通过 ELISA 测量活性基质金属蛋白酶 9、TNF-α、CXCL8 和 IL-10 的释放。
前两个最低密度的分数超过 90%为凋亡/坏死，其余分数超过 70%为存活。非吸烟者和吸烟者的巨噬细胞为 CD163（+）、CD206（+）、CD14（+）和 CD40（-），而 COPD 患者的巨噬细胞则不太明确，所有受体的表达均明显降低。受体表达与密度无关。与非吸烟者相比，密度为 1.036 至 1.048 g/mL 的 COPD 患者的巨噬细胞释放的活性基质金属蛋白酶 9 水平更高，而其余部分之间没有差异。与非吸烟者和吸烟者相比，来自 COPD 患者的密度为 1.036 至 1.048 g/mL 的巨噬细胞对 LPS 刺激的反应性较低。糖皮质激素不敏感是针对促炎细胞因子的，因为布地奈德对 LPS 刺激的 IL-10 释放的抑制作用对所有巨噬细胞都是相似的。
这项研究确定了一种在糖皮质激素不敏感的 COPD 患者肺部中存在的特定巨噬细胞表型，其密度为 1.036 至 1.048 g/mL，但与当前的巨噬细胞表型概念不对应。
