Reduced Clearance of Fungal Spores by Chronic Obstructive Pulmonary Disease GM-CSF- and M-CSF-derived Macrophages.
Reduced Clearance of Fungal Spores by Chronic Obstructive Pulmonary Disease GM-CSF- and M-CSF-derived Macrophages.
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慢性阻塞性肺病 GM-CSF 和 M-CSF 衍生巨噬细胞对真菌孢子的清除率降低。
DOI：
10.1165/rcmb.2017-0351le
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发表时间：
2018
影响因子：
6.4
通讯作者：
Wrench C
中科院分区：
文献类型：
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作者：
Wrench C
慢性阻塞性肺病 (COPD) 与由巨噬细胞、中性粒细胞和 T 淋巴细胞组成的特征性潜在炎症有关。尽管肺部先天免疫细胞数量增加，但慢性阻塞性肺病患者仍容易出现感染加重。这些主要归因于病毒和/或细菌感染；然而，人们越来越多地认识到真菌病原体是 COPD 恶化的诱因，烟曲霉是 COPD 稳定状态和恶化期间患者痰液中最常见的菌种 (1)。在之前的一项研究中，在约一半 COPD 受试者的痰液中培养了丝状烟曲霉，该丝状烟曲霉与使用较高剂量的吸入皮质类固醇有关 (2)。此外，对烟曲霉的过敏被证明与较差的肺功能和预后相关，尽管这些关系的确切性质需要进一步研究 (2, 3)。为什么这些真菌孢子持续存在于慢性阻塞性肺病患者的气道中尚不清楚，但这表明肺泡巨噬细胞无法清除。在 COPD 中，肺泡巨噬细胞对细菌和凋亡细胞的吞噬作用降低 (4, 5)，据推测这有助于疾病的发病机制。此外，这种缺陷也发生在与循环单核细胞分化的巨噬细胞（单核细胞衍生的巨噬细胞 [MDM]）中(5)，这表明这些细胞可用于模拟 COPD 中巨噬细胞功能的这一方面。这种缺陷是否延伸到烟曲霉等真菌孢子尚不清楚。因此，我们使用 MDM 模型，分别使用粒细胞-巨噬细胞集落刺激因子 (GM-CSF；G-Mw) 和 M-CSF (M-Mw)，将来自非吸烟者、吸烟者和 COPD 患者的单核细胞分化为促炎和分解巨噬细胞（数据补充中的表 E1）(6)。然后，我们测量了这些细胞对烟曲霉的吞噬潜力和细胞因子输出（有关详细信息，请参阅本文数据补充中的方法部分）。这些模型被认为代表了极化的巨噬细胞；因此，通过使用两种巨噬细胞表型，我们可以研究对烟曲霉的先天反应的任何变化是否是由于巨噬细胞表型的差异造成的，以及疾病状态是否导致了这些差异 (6, 7)。使用表达 GFP 的烟曲霉，我们证实 G-Mw（图 1A）和 M-Mw（图 1B）对分生孢子的摄取具有时间依赖性，但与来自健康对照的细胞相比，来自吸烟者和 COPD 患者的细胞显示吞噬这些孢子的细胞百分比显着降低。与慢性阻塞性肺病患者相比，吸烟者的细胞反应在任一巨噬细胞表型上均无统计学差异。我们还通过测量每个细胞摄取的分生孢子数量（通过中值荧光强度测量；图E1A和E1B）来评估每个细胞的吞噬能力，并观察到吸烟者和慢性阻塞性肺病患者的细胞有类似的抑制反应。通过共聚焦显微镜证实了分生孢子的内化（图1C和1D）。年龄、吸烟史或任何肺功能参数与 G-Mw 或 M-Mw 摄取分生孢子之间没有观察到相关性。然而，当比较同一患者的细胞时，G-Mw 和 M-Mw 的吞噬作用之间存在微弱的正相关性（图 E1E）。这些数据表明真菌孢子的清除是......
Chronic obstructive pulmonary disease (COPD) is associated with a characteristic underlying inflammation that comprises macrophages, neutrophils, and T-lymphocytes. Despite the presence of increased numbers of pulmonary innate immune cells, patients with COPD are prone to infective exacerbations. These are predominantly attributed to viral and/or bacterial infection; however, fungal pathogens are increasingly being recognized as contributors to COPD exacerbations, with Aspergillus fumigatus being the most common species detected in the sputum of patients with COPD both at steady state and during exacerbation (1). In a previous study, filamentous A. fumigatus was cultured in sputum from approximately half of the subjects with COPD and was associated with the use of higher doses of inhaled corticosteroid (2). In addition, sensitization to A. fumigatus was shown to correlate with poorer lung function and prognosis, although the exact nature of these relationships requires further study (2, 3). Why these fungal spores persist in the airways of patients with COPD is not known, but it suggests a failure of clearance by alveolar macrophages. In COPD, alveolar macrophages exhibit reduced phagocytosis of bacteria and apoptotic cells (4, 5), and it is postulated that this contributes to the pathogenesis of disease. Furthermore, this defect also occurs in macrophages that have been differentiated from circulating monocytes (monocyte-derived macrophages [MDMs])(5), suggesting that these cells can be used to model this aspect of macrophage function in COPD. Whether this defect extends to fungal spores such as A. fumigatus is unknown. Therefore, we used the MDM model to differentiate monocytes into proinflammatory and resolving macrophages from nonsmokers, smokers, and patients with COPD (Table E1 in the data supplement), using granulocyte–macrophage colony–stimulating factor (GM-CSF; G-Mw) and M-CSF (M-Mw), respectively (6). We then measured the phagocytic potential and cytokine output of these cells in response to A. fumigatus (for details, see this article’s METHODS section in the data supplement). These models are considered to represent polarized macrophages; therefore, by using both macrophage phenotypes, one can investigate whether any changes in the innate response to A. fumigatus are due to differences in the macrophage phenotype, and whether the disease state is contributing to these differences (6, 7). Using GFP-expressing A. fumigatus, we confirmed that uptake of conidia by both G-Mw (Figure 1A) and M-Mw (Figure 1B) was time dependent, but that cells derived from smokers and patients with COPD showed a significant reduction in the percentage of cells that had phagocytosed these spores compared with cells derived from healthy controls. There was no statistical difference in the responses of cells from smokers compared with patients with COPD in either macrophage phenotype. We also assessed the capacity of each cell to phagocytose by measuring the number of conidia taken up per cell (measured by median fluorescence intensity; Figures E1A and E1B), and observed similar suppressed responses by cells from smokers and patients with COPD. Internalization of conidia was confirmed by confocal microscopy (Figures 1C and 1D). No correlations were observed between age, smoking history, or any lung-function parameters and the uptake of conidia by either G-Mw or M-Mw. However, when comparing cells from the same patient, there was a weak positive correlation between phagocytosis of G-Mw and that of M-Mw (Figure E1E). These data suggest that the clearance of fungal spores is …
DOI：
10.1146/annurev-pathol-052016-100342
发表时间：
2017-01-24
期刊：
Annual review of pathology
影响因子：
--
作者：
Wheeler ML;Limon JJ;Underhill DM
通讯作者：
Underhill DM
影响因子：
5.8
作者：
Huerta A;Soler N;Esperatti M;Guerrero M;Menendez R;Gimeno A;Zalacaín R;Mir N;Aguado JM;Torres A
通讯作者：
Torres A