CFTR-mediated halide transport in phagosomes of human neutrophils

Richard G. Painter, Luis Marrero, Gisele A. Lombard, Vincent G. Valentine, William M. Nauseef, Guoshun Wang

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Chloride serves as a critical component of innate host defense against infection, providing the substrate for MPO-catalyzed production of HOCl in the phagosome of human neutrophils. Here, we used halide-specific fluorescent sensors covalently coupled to zymosan particles to investigate the kinetics of chloride and iodide transport in phagosomes of human neutrophils. Using the self-ratioable fluorescent probe specific for chloride anion, we measured chloride dynamics within phagosomes in response to extracellular chloride changes by quantitative fluorescence microscopy. Under the experimental conditions used, normal neutrophils showed rapid phagosomal chloride uptake with an initial influx rate of 0.31 ± 0.04 mM/s (n=5). GlyH-101, a CFTR inh, decreased the rate of uptake in a dose-dependent manner. Neutrophils isolated from CF patients showed a significantly slower rate of chloride uptake by phagosomes, having an initial influx rate of 0.043 ± 0.012 mM/s (n=5). Interestingly, the steady-state level of chloride in CF phagosomes was ∼26 mM, significantly lower than that of the control (∼68 mM). As CFTR transports chloride as well as other halides, we conjugated an iodide-sensitive probe as an independent approach to confirm the results. The dynamics of iodide uptake by neutrophil phagosomes were monitored by flow cytometry. CFTRinh172 blocked 40-50% of the overall iodide uptake by phagosomes in normal neutrophils. In a parallel manner, the level of iodide uptake by CF phagosomes was only 20-30% of that of the control. Taken together, these results implicate CFTR in transporting halides into the phagosomal lumen.

Original languageEnglish (US)
Pages (from-to)933-942
Number of pages10
JournalJournal of Leukocyte Biology
Volume87
Issue number5
DOIs
StatePublished - May 2010

Fingerprint

Phagosomes
Chlorides
Neutrophils
Iodides
Zymosan
Fluorescent Dyes
Fluorescence Microscopy
Anions
Flow Cytometry

Keywords

  • Chloride sensor
  • Fluorescent halide probes
  • Hypochlorous acid
  • Iodide sensor
  • Ion transport

ASJC Scopus subject areas

  • Cell Biology
  • Immunology

Cite this

Painter, R. G., Marrero, L., Lombard, G. A., Valentine, V. G., Nauseef, W. M., & Wang, G. (2010). CFTR-mediated halide transport in phagosomes of human neutrophils. Journal of Leukocyte Biology, 87(5), 933-942. https://doi.org/10.1189/jlb.1009655

CFTR-mediated halide transport in phagosomes of human neutrophils. / Painter, Richard G.; Marrero, Luis; Lombard, Gisele A.; Valentine, Vincent G.; Nauseef, William M.; Wang, Guoshun.

In: Journal of Leukocyte Biology, Vol. 87, No. 5, 05.2010, p. 933-942.

Research output: Contribution to journalArticle

Painter, RG, Marrero, L, Lombard, GA, Valentine, VG, Nauseef, WM & Wang, G 2010, 'CFTR-mediated halide transport in phagosomes of human neutrophils', Journal of Leukocyte Biology, vol. 87, no. 5, pp. 933-942. https://doi.org/10.1189/jlb.1009655
Painter RG, Marrero L, Lombard GA, Valentine VG, Nauseef WM, Wang G. CFTR-mediated halide transport in phagosomes of human neutrophils. Journal of Leukocyte Biology. 2010 May;87(5):933-942. https://doi.org/10.1189/jlb.1009655
Painter, Richard G. ; Marrero, Luis ; Lombard, Gisele A. ; Valentine, Vincent G. ; Nauseef, William M. ; Wang, Guoshun. / CFTR-mediated halide transport in phagosomes of human neutrophils. In: Journal of Leukocyte Biology. 2010 ; Vol. 87, No. 5. pp. 933-942.
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