RNA interference against CFTR affects HL60-derived neutrophil microbicidal function

Free Radic Biol Med. 2010 Dec 15;49(12):1872-80. doi: 10.1016/j.freeradbiomed.2010.09.012. Epub 2010 Sep 24.

Abstract

Biosynthesis of hypochlorous acid, a potent antimicrobial oxidant, in phagosomes is one of the chief mechanisms employed by polymorphonuclear neutrophils to combat infections. This reaction, catalyzed by myeloperoxidase, requires chloride anion (Cl(-)) as a substrate. Thus, Cl(-) availability is a rate-limiting factor that affects neutrophil microbicidal function. Our previous research demonstrated that defective CFTR, a cAMP-activated chloride channel, present in cystic fibrosis (CF) patients leads to deficient chloride transport to neutrophil phagosomes and impaired bacterial killing. To confirm this finding, here we used RNA interference against this chloride channel to abate CFTR expression in the neutrophil-like cells derived from HL60 cells, a promyelocytic leukemia cell line, with dimethyl sulfoxide. The resultant CFTR deficiency in the phagocytes compromised their bactericidal capability, thereby recapitulating the phenotype seen in CF patient cells. The results provide further evidence suggesting that CFTR plays an important role in phagocytic host defense.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation / genetics
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism
  • HL-60 Cells
  • Humans
  • Hydrogen Peroxide / metabolism
  • Hypochlorous Acid / metabolism
  • Microbial Viability
  • Models, Biological
  • Neutrophils / physiology*
  • Phagocytes / physiology*
  • Phagocytosis
  • Pseudomonas aeruginosa / physiology*
  • RNA Interference*
  • RNA, Small Interfering / metabolism
  • Superoxides / metabolism

Substances

  • CFTR protein, human
  • RNA, Small Interfering
  • Superoxides
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Hypochlorous Acid
  • Hydrogen Peroxide