Hodge Sandra, Tran Hai B, Hamon Rhys, Roscioli Eugene, Hodge Greg, Jersmann Hubertus, Ween Miranda, Reynolds Paul N, Yeung Arthur, Treiberg Jennifer, Wilbert Sibylle
Lung Research Unit, Hanson Institute, and Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, Australia;
Department of Medicine, University of Adelaide, Adelaide, Australia; and.
Am J Physiol Lung Cell Mol Physiol. 2017 May 1;312(5):L678-L687. doi: 10.1152/ajplung.00518.2016. Epub 2017 Mar 3.
We reported defective efferocytosis associated with cigarette smoking and/or airway inflammation in chronic lung diseases, including chronic obstructive pulmonary disease, severe asthma, and childhood bronchiectasis. We also showed defects in phagocytosis of nontypeable (NTHi), a common colonizer of the lower airway in these diseases. These defects could be substantially overcome with low-dose azithromycin; however, chronic use may induce bacterial resistance. The aim of the present study was therefore to investigate two novel macrolides-2'-desoxy-9-(S)-erythromycylamine (GS-459755) and azithromycin-based 2'-desoxy molecule (GS-560660)-with significantly diminished antibiotic activity against , , , and We tested their effects on efferocytosis, phagocytosis of NTHi, cell viability, receptors involved in recognition of apoptotic cells and/or NTHi (flow cytometry), secreted and cleaved intracellular IL-1β (cytometric bead array, immunofluorescence/confocal microscopy), and nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) using primary alveolar macrophages and THP-1 macrophages ± 10% cigarette smoke extract. Dose-response experiments showed optimal prophagocytic effects of GS-459755 and GS-560660 at concentrations of 0.5-1 µg/ml compared with our findings with azithromycin. Both macrolides significantly improved phagocytosis of apoptotic cells and NTHi (e.g., increases in efferocytosis and phagocytosis of NTHi: GS-459755, 23 and 22.5%, = 0.043; GS-560660, 23.5 and 22%, = 0.043, respectively). Macrophage viability remained >85% following 24 h exposure to either macrolide at concentrations up to 20 µg/ml. Secreted and intracellular-cleaved IL-1β was decreased with both macrolides with no significant changes in recognition molecules c-mer proto-oncogene tyrosine kinase; scavenger receptor class A, member 1; Toll-like receptor 2/4; or CD36. Particulate cytoplasmic immunofluorescence of NLRP3 inflammasome was also reduced significantly. We conclude that GS-459755 and GS-560660 may be useful for reducing airway inflammation in chronic lung diseases without inducing bacterial resistance.
我们报告了在慢性肺部疾病（包括慢性阻塞性肺疾病、重度哮喘和儿童支气管扩张症）中，与吸烟和/或气道炎症相关的吞噬细胞清除功能缺陷。我们还发现这些疾病的下呼吸道常见定植菌——不可分型流感嗜血杆菌（NTHi）的吞噬功能存在缺陷。低剂量阿奇霉素可显著克服这些缺陷；然而，长期使用可能会诱导细菌耐药性。因此，本研究的目的是研究两种新型大环内酯类药物——2'-脱氧-9-(S)-红霉素胺（GS-459755）和基于阿奇霉素的2'-脱氧分子（GS-560660），它们对肺炎链球菌、金黄色葡萄球菌、流感嗜血杆菌和卡他莫拉菌的抗生素活性显著降低。我们使用原代肺泡巨噬细胞和THP-1巨噬细胞±10%香烟烟雾提取物，测试了它们对吞噬细胞清除功能、NTHi吞噬功能、细胞活力、参与识别凋亡细胞和/或NTHi的受体（流式细胞术）、分泌和裂解的细胞内白细胞介素-1β（细胞因子珠阵列、免疫荧光/共聚焦显微镜）以及含核苷酸结合寡聚化结构域样受体家族吡啉结构域3（NLRP3）的影响。剂量反应实验表明，与阿奇霉素相比，GS-459755和GS-560660在浓度为0.5-1μg/ml时具有最佳的促吞噬作用。两种大环内酯类药物均显著改善了凋亡细胞和NTHi的吞噬功能（例如，NTHi的吞噬细胞清除功能和吞噬功能增加：GS-459755分别为23%和22.5%，P = 0.043；GS-560660分别为23.5%和22%，P = 0.043）。在浓度高达20μg/ml的情况下，巨噬细胞暴露于任何一种大环内酯类药物24小时后，细胞活力仍>85%。两种大环内酯类药物均降低了分泌型和细胞内裂解型白细胞介素-1β，而识别分子c-mer原癌基因酪氨酸激酶、A类清道夫受体成员1、Toll样受体2/4或CD36无显著变化。NLRP3炎性小体的颗粒状细胞质免疫荧光也显著降低。我们得出结论，GS-459755和GS-560660可能有助于减少慢性肺部疾病中的气道炎症，而不会诱导细菌耐药性。
