Altered Degranulation and pH of Neutrophil Phagosomes Impacts Antimicrobial Efficiency in Cystic Fibrosis.
Altered Degranulation and pH of Neutrophil Phagosomes Impacts Antimicrobial Efficiency in Cystic Fibrosis.
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DOI：
10.3389/fimmu.2020.600033
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发表时间：
2020
影响因子：
7.3
通讯作者：
Reeves EP
中科院分区：
文献类型：
--
作者：
Hayes E;Murphy MP;Pohl K;Browne N;McQuillan K;Saw LE;Foley C;Gargoum F;McElvaney OJ;Hawkins P;Gunaratnam C;McElvaney NG;Reeves EP
研究致力于了解囊性纤维化 (PWCF) 患者中性粒细胞抗菌能力受损的原因。本研究的目的是关注细菌吞噬体。评估了细胞质颗粒脱颗粒和 pH 值变化的可能变化。从 PWCF (n = 28)、接受 ivacaftor 治疗的 PWCF (n = 10) 和健康对照 (n = 28) 中纯化循环中性粒细胞。通过蛋白质印迹分析和流式细胞术评估脱颗粒。使用 BCECF-AM 标记的金黄色葡萄球菌或 SNARF 标记的白色念珠菌测定吞噬体的 pH 值。所有测试处理的抗菌效果均通过菌落形成单位计数来确定。 CF 和健康对照中性粒细胞的细菌杀灭效果存在差异 (p = 0.0006)。通过使用流式细胞术和亚细胞分级分离，发现 CF 吞噬体中吞噬体内脱颗粒的动力学显着改变，初级颗粒 CD63 (p = 0.0001) 和髓过氧化物酶 (MPO) 含量 (p = 0.03) 增加证明了这一点。相反，观察到二级和三级颗粒 CD66b 减少（p = 0.002）以及 hCAP-18 和 MMP-9 减少（p = 0.02）。 8 分钟吞噬作用后，PWCF 中性粒细胞吞噬体的 pH 值显着升高 (p = 0.0001)，并且与 HC 相比，活细菌的百分比显着增加 (p = 0.002)。结果表明，记录的吞噬体 pH 值的变化为 MPO 相关过氧化物酶、α-防御素和 azurocidine 酶促杀灭金黄色葡萄球菌和铜绿假单胞菌产生了次优条件。使用离子通道增强剂 ivacaftor 治疗后，CF 中性粒细胞中 MPO 脱粒失调 (p = 0.02) 和延长的吞噬体碱化模式在体内正常化 (p = 0.04)。我们的结果证实，PWCF 循环中性粒细胞的改变可以通过 CFTR 调节剂治疗得到纠正，并提出与 CF 中质子通道活性可能延迟相关的问题。
Studies have endeavored to understand the cause for impaired antimicrobial killing by neutrophils of people with cystic fibrosis (PWCF). The aim of this study was to focus on the bacterial phagosome. Possible alterations in degranulation of cytoplasmic granules and changes in pH were assessed. Circulating neutrophils were purified from PWCF (n = 28), PWCF receiving ivacaftor therapy (n = 10), and healthy controls (n = 28). Degranulation was assessed by Western blot analysis and flow cytometry. The pH of phagosomes was determined by use of BCECF-AM-labelled Staphylococcus aureus or SNARF labelled Candida albicans. The antibacterial effect of all treatments tested was determined by colony forming units enumeration. Bacterial killing by CF and healthy control neutrophils were found to differ (p = 0.0006). By use of flow cytometry and subcellular fractionation the kinetics of intraphagosomal degranulation were found to be significantly altered in CF phagosomes, as demonstrated by increased primary granule CD63 (p = 0.0001) and myeloperoxidase (MPO) content (p = 0.03). In contrast, decreased secondary and tertiary granule CD66b (p = 0.002) and decreased hCAP-18 and MMP-9 (p = 0.02), were observed. After 8 min phagocytosis the pH in phagosomes of neutrophils of PWCF was significantly elevated (p = 0.0001), and the percentage of viable bacteria was significantly increased compared to HC (p = 0.002). Results demonstrate that the recorded alterations in phagosomal pH generate suboptimal conditions for MPO related peroxidase, and α-defensin and azurocidine enzymatic killing of Staphylococcus aureus and Pseudomonas aeruginosa. The pattern of dysregulated MPO degranulation (p = 0.02) and prolonged phagosomal alkalinization in CF neutrophils were normalized in vivo following treatment with the ion channel potentiator ivacaftor (p = 0.04). Our results confirm that alterations of circulating neutrophils from PWCF are corrected by CFTR modulator therapy, and raise a question related to possible delayed proton channel activity in CF.
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从人类嗜中性粒细胞分离的功能活性吞噬体中的氯化物转运。
影响因子：
7.4
作者：
Aiken, Martha L.;Painter, Richard G.;Zhou, Yun;Wang, Guoshun
通讯作者：
Wang, Guoshun
影响因子：
17.1
作者：
Bergin, David A.;Reeves, Emer P.;McElvaney, Noel G.
通讯作者：
McElvaney, Noel G.
影响因子：
5.5
作者：
Ng HP;Jennings S;Wellems D;Sun F;Xu J;Nauseef WM;Wang G
通讯作者：
Wang G
DOI：
10.1164/ajrccm.150.1.7912987
发表时间：
1994-07-01
影响因子：
24.7
作者：
BIRRER, P;MCELVANEY, NG;CRYSTAL, RG
通讯作者：
CRYSTAL, RG
影响因子：
3.1
作者：
Hampton, MB;Kettle, AJ;Winterbourn, CC
通讯作者：
Winterbourn, CC