Winkler Aaron R, Nocka Karl H, Sulahian Timothy H, Kobzik Lester, Williams Cara M M
Wyeth Research, Cambridge, Massachusetts 02140, USA.
Exp Lung Res. 2008 Nov;34(9):599-629. doi: 10.1080/01902140802366261.
Pulmonary macrophages (MØs) are essential for clearance of inhaled particles, innate immunity, and lung tissue maintenance. However, the products of activated MØs have also been implicated in inflammation and tissue destruction, including in chronic obstructive pulmonary disease (COPD). Primary human alveolar macrophages (AMs) are available in limited numbers via bronchoalveolar lavage (BAL) or sputum induction, and BAL macrophages are not commonly available to all researchers. A readily available, plentiful, but representative surrogate for AMs would advance understanding of the contribution of macrophages to lung pathophysiology. Herein the authors describe a method for the in vitro derivation of AM-like cells using primary human peripheral blood monocytes differentiated in suspension with granulocyte-macrophage colony-stimulating factor (GM-CSF). The method produces a cell population with a consistent and stable phenotype. Flow cytometry reveals that GM-CSF-derived macrophages (GM-MØs) express lineage markers, immunoglobulin gamma (Fc gamma) receptors, adhesion molecules, antigen presentation coreceptors, and scavenger receptors akin to AMs. Functionally, cigarette smoke activates extracellular signal-related kinase (ERK) and p38 mitogen-activated protein (MAP) kinase, enhances interleukin 8 (IL8) production from GM-MØs and inhibits phagocytosis, phenotypes previously described for smokers' AMs. Global transcriptional profiling revealed significant overlap in regulated genes between smokers' AMs and GM-MØs treated with cigarette smoke preparations in vitro.
肺巨噬细胞（MØs）对于清除吸入颗粒、固有免疫和维持肺组织至关重要。然而，活化的MØs产物也与炎症和组织破坏有关，包括在慢性阻塞性肺疾病（COPD）中。通过支气管肺泡灌洗（BAL）或痰液诱导获得的原代人肺泡巨噬细胞（AMs）数量有限，并且并非所有研究人员都能普遍获得BAL巨噬细胞。一种易于获得、数量丰富且具有代表性的AMs替代物将有助于深入了解巨噬细胞对肺病理生理学的贡献。在此，作者描述了一种使用在悬浮状态下用粒细胞-巨噬细胞集落刺激因子（GM-CSF）分化的原代人外周血单核细胞体外诱导产生AMs样细胞的方法。该方法产生的细胞群体具有一致且稳定的表型。流式细胞术显示，GM-CSF衍生的巨噬细胞（GM-MØs）表达谱系标志物、免疫球蛋白γ（Fcγ）受体、黏附分子、抗原呈递共受体和类似于AMs的清道夫受体。在功能上，香烟烟雾激活细胞外信号相关激酶（ERK）和p38丝裂原活化蛋白（MAP）激酶，增强GM-MØs产生白细胞介素8（IL8）并抑制吞噬作用，这些表型先前在吸烟者的AMs中已有描述。全基因组转录谱分析显示，吸烟者的AMs与体外经香烟烟雾制剂处理的GM-MØs之间在受调控基因方面存在显著重叠。
