CFTR expression in human neutrophils and the phagolysosomal chlorination defect in cystic fibrosis

Richard G. Painter, Vincent G. Valentine, Nicholas A. Lanson, Kevin Leidal, Qiang Zhang, Gisele Lombard, Connie Thompson, Anand Viswanathan, William M. Nauseef, Guangdi Wang, Guoshun Wang

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Abstract

Production of hypochlorous acid (HOCI) in neutrophils, a critical oxidant involved in bacterial killing, requires chloride anions. Because the primary defect of cystic fibrosis (CF) is the loss of chloride transport function of the CF transmembrane conductance regulator (CFTR), we hypothesized that CF neutrophils may be deficient in chlorination of bacterial components due to a limited chloride supply to the phagolysosomal compartment. Multiple approaches, including RT-PCR, immunofluorescence staining, and immunoblotting, were used to demonstrate that CFTR is expressed in resting neutrophils at the mRNA and protein levels. Probing fractions of resting neutrophils isolated by Percoll gradient fractionation and free flow electropheresis for CFTR revealed its presence exclusively in secretory vesicles. The CFTR chloride channel was also detected in phagolysosomes, a special organelle formed after phagocytosis. Interestingly, HL-60 cells, a human promyelocytic leukemia cell line, upregulated CFTR expresssion when induced to differentiate into neutrophils with DMSO, strongly suggesting its potential role in mature neutrophil function. Analyses by gas chromatography and mass spectrometry (GC-MS) revealed that neutrophils from CF patients had a defect in their ability to chlorinate bacterial proteins from Pseudomonas aeruginosa metabolically prelabeled with [13C]-L-tyrosine, unveiling defective intraphagolysosomal HOCI production. In contrast, both normal and CF neutrophils exhibited normal extracellular production of HOCI when stimulated with phorbol ester, indicating that CF neutrophils had the normal ability to produce this oxidant in the extracellular medium. This report provides evidence which suggests that CFTR channel expression in neutrophils and its dysfunction affect neutrophil chlorination of phagocytosed bacteria.

Original languageEnglish (US)
Pages (from-to)10260-10269
Number of pages10
JournalBiochemistry
Volume45
Issue number34
DOIs
StatePublished - Aug 29 2006
Externally publishedYes

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Hypochlorous Acid
Chlorination
Halogenation
Cystic Fibrosis
Chlorides
Neutrophils
Oxidants
Defects
Cystic Fibrosis Transmembrane Conductance Regulator
Chloride Channels
Bacterial Proteins
Phorbol Esters
Fractionation
Dimethyl Sulfoxide
Gas chromatography
Anions
Mass spectrometry
Tyrosine
Bacteria
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Painter, R. G., Valentine, V. G., Lanson, N. A., Leidal, K., Zhang, Q., Lombard, G., ... Wang, G. (2006). CFTR expression in human neutrophils and the phagolysosomal chlorination defect in cystic fibrosis. Biochemistry, 45(34), 10260-10269. https://doi.org/10.1021/bi060490t

CFTR expression in human neutrophils and the phagolysosomal chlorination defect in cystic fibrosis. / Painter, Richard G.; Valentine, Vincent G.; Lanson, Nicholas A.; Leidal, Kevin; Zhang, Qiang; Lombard, Gisele; Thompson, Connie; Viswanathan, Anand; Nauseef, William M.; Wang, Guangdi; Wang, Guoshun.

In: Biochemistry, Vol. 45, No. 34, 29.08.2006, p. 10260-10269.

Research output: Contribution to journalArticle

Painter, RG, Valentine, VG, Lanson, NA, Leidal, K, Zhang, Q, Lombard, G, Thompson, C, Viswanathan, A, Nauseef, WM, Wang, G & Wang, G 2006, 'CFTR expression in human neutrophils and the phagolysosomal chlorination defect in cystic fibrosis', Biochemistry, vol. 45, no. 34, pp. 10260-10269. https://doi.org/10.1021/bi060490t
Painter RG, Valentine VG, Lanson NA, Leidal K, Zhang Q, Lombard G et al. CFTR expression in human neutrophils and the phagolysosomal chlorination defect in cystic fibrosis. Biochemistry. 2006 Aug 29;45(34):10260-10269. https://doi.org/10.1021/bi060490t
Painter, Richard G. ; Valentine, Vincent G. ; Lanson, Nicholas A. ; Leidal, Kevin ; Zhang, Qiang ; Lombard, Gisele ; Thompson, Connie ; Viswanathan, Anand ; Nauseef, William M. ; Wang, Guangdi ; Wang, Guoshun. / CFTR expression in human neutrophils and the phagolysosomal chlorination defect in cystic fibrosis. In: Biochemistry. 2006 ; Vol. 45, No. 34. pp. 10260-10269.
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