Pohl Kerstin, Hayes Elaine, Keenan Joanne, Henry Michael, Meleady Paula, Molloy Kevin, Jundi Bakr, Bergin David A, McCarthy Cormac, McElvaney Oliver J, White Michelle M, Clynes Martin, Reeves Emer P, McElvaney Noel G
Respiratory Research Division, Department of Medicine, Royal College of Surgeons in Ireland, Education and Research Centre, Beaumont Hospital, Dublin, Ireland; and.
National Institute for Cellular Biotechnology, Dublin City University, Dublin, Ireland.
Blood. 2014 Aug 14;124(7):999-1009. doi: 10.1182/blood-2014-02-555268. Epub 2014 Jun 16.
Studies have endeavored to reconcile whether dysfunction of neutrophils in people with cystic fibrosis (CF) is a result of the genetic defect or is secondary due to infection and inflammation. In this study, we illustrate that disrupted function of the CF transmembrane conductance regulator (CFTR), such as that which occurs in patients with ∆F508 and/or G551D mutations, correlates with impaired degranulation of antimicrobial proteins. We demonstrate that CF blood neutrophils release less secondary and tertiary granule components compared with control cells and that activation of the low-molecular-mass GTP-binding protein Rab27a, involved in the regulation of granule trafficking, is defective. The mechanism leading to impaired degranulation involves altered ion homeostasis caused by defective CFTR function with increased cytosolic levels of chloride and sodium, yet decreased magnesium measured in CF neutrophils. Decreased magnesium concentration in vivo and in vitro resulted in significantly decreased levels of GTP-bound Rab27a. Treatment of G551D patients with the ion channel potentiator ivacaftor resulted in normalized neutrophil cytosolic ion levels and activation of Rab27a, thereby leading to increased degranulation and bacterial killing. Our results confirm that intrinsic alterations of circulating neutrophils from patients with CF are corrected by ivacaftor, thus illustrating additional clinical benefits for CFTR modulator therapy.
多项研究致力于阐明囊性纤维化(CF)患者中性粒细胞功能障碍是基因缺陷所致,还是由感染和炎症继发引起。在本研究中,我们表明CF跨膜电导调节因子(CFTR)功能紊乱,如发生在携带∆F508和/或G551D突变患者中的情况,与抗菌蛋白脱颗粒受损相关。我们证明,与对照细胞相比,CF患者血液中的中性粒细胞释放的次级和三级颗粒成分较少,且参与颗粒运输调节的低分子量GTP结合蛋白Rab27a的激活存在缺陷。导致脱颗粒受损的机制涉及CFTR功能缺陷引起的离子稳态改变,CF中性粒细胞中氯化物和钠的胞质水平升高,而镁含量降低。体内和体外镁浓度降低导致结合GTP的Rab27a水平显著下降。用离子通道增强剂依伐卡托治疗G551D患者可使中性粒细胞胞质离子水平正常化,并激活Rab27a,从而导致脱颗粒增加和细菌杀伤作用增强。我们的结果证实,依伐卡托可纠正CF患者循环中性粒细胞的内在改变,从而说明CFTR调节剂治疗具有更多临床益处。