コンテンツへスキップ
Merck
  • A neutrophil intrinsic impairment affecting Rab27a and degranulation in cystic fibrosis is corrected by CFTR potentiator therapy.

A neutrophil intrinsic impairment affecting Rab27a and degranulation in cystic fibrosis is corrected by CFTR potentiator therapy.

Blood (2014-06-18)
Kerstin Pohl, Elaine Hayes, Joanne Keenan, Michael Henry, Paula Meleady, Kevin Molloy, Bakr Jundi, David A Bergin, Cormac McCarthy, Oliver J McElvaney, Michelle M White, Martin Clynes, Emer P Reeves, Noel G McElvaney
要旨

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.

材料
製品番号
ブランド
製品内容

Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
塩化ナトリウム, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
塩化マグネシウム 溶液, for molecular biology, 1.00 M±0.01 M
Sigma-Aldrich
塩化マグネシウム, anhydrous, ≥98%
Sigma-Aldrich
塩化カルシウム 溶液, BioUltra, for molecular biology, ~1 M in H2O
Sigma-Aldrich
塩化ナトリウム 溶液, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
塩化ナトリウム 溶液, 0.9% in water, BioXtra, suitable for cell culture
Sigma-Aldrich
塩化ナトリウム, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
塩化カルシウム, anhydrous, BioReagent, suitable for insect cell culture, suitable for plant cell culture, ≥96.0%
Sigma-Aldrich
HEPES, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
HEPES緩衝液, 1 M in H2O
Sigma-Aldrich
塩化マグネシウム, powder, <200 μm
Sigma-Aldrich
塩化ナトリウム, JIS special grade, ≥99.5%
SAFC
HEPES
Sigma-Aldrich
塩化ナトリウム 溶液, BioUltra, for molecular biology, ~5 M in H2O
Sigma-Aldrich
塩化カルシウム, anhydrous, powder, 99.99% trace metals basis
Sigma-Aldrich
塩化ナトリウム, BioUltra, for molecular biology, ≥99.5% (AT)
Sigma-Aldrich
塩化ナトリウム, 99.999% trace metals basis
Sigma-Aldrich
塩化マグネシウム 溶液, BioUltra, for molecular biology, 2 M in H2O
Sigma-Aldrich
塩化マグネシウム, BioReagent, suitable for insect cell culture, ≥97.0%
Sigma-Aldrich
イソフタル酸, 99%
Sigma-Aldrich
HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)
Sigma-Aldrich
塩化ナトリウム, BioXtra, ≥99.5% (AT)
SAFC
HEPES
Sigma-Aldrich
塩化ナトリウム, SAJ first grade, ≥99.0%
Sigma-Aldrich
塩化カルシウム, AnhydroBeads, −10 mesh, ≥99.9% trace metals basis
Sigma-Aldrich
塩化マグネシウム 溶液, PCR Reagent, 25 mM MgCI2 solution for PCR
Sigma-Aldrich
塩化マグネシウム, AnhydroBeads, −10 mesh, 99.9% trace metals basis
Sigma-Aldrich
HEPES, BioXtra, pH 5.0-6.5 (1 M in H2O), ≥99.5% (titration)