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Role of CFTR expressed by neutrophils in modulating acute lung inflammation and injury in mice.

Su X, Looney MR, Su HE, Lee JW, Song Y, Matthay MA - Inflamm. Res. (2011)

Bottom Line: Cystic fibrosis transmembrane conductance regulator (CFTR) regulates infection and inflammation.Wild-type mice were reconstituted with F508del neutrophils or bone marrow and then intratracheally challenged with LPS to observe lung inflammatory response.Pharmacologic inhibition and genetic mutation of CFTR in neutrophils activated NF-κB and facilitated macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-α (TNF-α) production.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Research Institute, University of California, San Francisco, HSW 825, 505 Parnassus AVE, San Francisco, CA 94143-0130, USA. suxiao1@yahoo.com

ABSTRACT

Objective and design: Cystic fibrosis transmembrane conductance regulator (CFTR) regulates infection and inflammation. In this study, we investigated whether a lack of functional CFTR in neutrophils would promote lipopolysaccharide (LPS)-induced lung inflammation and injury.

Materials and methods: CFTR-inhibited or F508del-CFTR-mutated neutrophils were stimulated with LPS and cultured to evaluate production of cytokines and NF-κB activation. Wild-type mice were reconstituted with F508del neutrophils or bone marrow and then intratracheally challenged with LPS to observe lung inflammatory response.

Results: Pharmacologic inhibition and genetic mutation of CFTR in neutrophils activated NF-κB and facilitated macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-α (TNF-α) production. Wild-type mice reconstituted with F508del neutrophils and bone marrow had more severe lung inflammation and injury after LPS challenge compared to wild-type mice receiving wild-type neutrophils or bone marrow reconstitution.

Conclusions: Lack of functional CFTR in neutrophils can promote LPS-induced acute lung inflammation and injury.

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Related in: MedlinePlus

Pro- and anti-inflammatory cytokines in neutrophils balance is impaired without functional CFTR. a–c Production of MIP-2 (a) and interleukin-10 (b) and NF-κB activation (c) in the cultured bone-marrow derived wild-type neutrophils pretreated with CFTRinh-172 and then stimulated with LPS at 12 h. **P < 0.01 for LPS + CFTRinh-172 vs LPS only. d–f Production of MIP-2 (d) and interleukin-10 (e) and NF-κB activation (f) in the cultured bone-marrow derived wild-type neutrophils and F508del neutrophils simulated with LPS at 12 h. *P < 0.05, **P < 0.01 for wild-type neutrophils receiving LPS vs F508del neutrophils stimulated with LPS. N = 6–8 in each group. Values are mean ± SD
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Fig4: Pro- and anti-inflammatory cytokines in neutrophils balance is impaired without functional CFTR. a–c Production of MIP-2 (a) and interleukin-10 (b) and NF-κB activation (c) in the cultured bone-marrow derived wild-type neutrophils pretreated with CFTRinh-172 and then stimulated with LPS at 12 h. **P < 0.01 for LPS + CFTRinh-172 vs LPS only. d–f Production of MIP-2 (d) and interleukin-10 (e) and NF-κB activation (f) in the cultured bone-marrow derived wild-type neutrophils and F508del neutrophils simulated with LPS at 12 h. *P < 0.05, **P < 0.01 for wild-type neutrophils receiving LPS vs F508del neutrophils stimulated with LPS. N = 6–8 in each group. Values are mean ± SD

Mentions: To study whether lack of functional CFTR in neutrophils affects both pro- and anti-inflammatory cytokines, isolated bone marrow neutrophils were pretreated with CFTRinh-172 (5 μM), stimulated with LPS, and cultured for 12 h to measure MIP-2 and IL-10 in the medium. The adherent neutrophils were then harvested for measurement of p65 NF-κB subunit in nuclear extract; MIP-2 levels increased more than two-fold (Fig. 4a) and IL-10 was significantly decreased (Fig. 4b) in the medium. The p65 NF-κB subunit in the neutrophil nucleus extract with LPS plus CFTRinh-172 inhibition was increased two-fold compared to LPS only (Fig. 4c). To confirm this result, F508del and wild-type neutrophils were cultured and stimulated with LPS for 12 h to measure MIP-2 and IL-10 in the medium. The cells were then harvested for measurement of p65 NF-κB in the nucleus. MIP-2 levels were elevated three-fold (Fig. 4d) and IL-10 was reduced (Fig. 4e). There was increased p65 NF-κB translocation to the nucleus in the F508del neutrophils stimulated with LPS compared to wild-type neutrophils receiving LPS challenge (Fig. 4f).Fig. 4


Role of CFTR expressed by neutrophils in modulating acute lung inflammation and injury in mice.

Su X, Looney MR, Su HE, Lee JW, Song Y, Matthay MA - Inflamm. Res. (2011)

Pro- and anti-inflammatory cytokines in neutrophils balance is impaired without functional CFTR. a–c Production of MIP-2 (a) and interleukin-10 (b) and NF-κB activation (c) in the cultured bone-marrow derived wild-type neutrophils pretreated with CFTRinh-172 and then stimulated with LPS at 12 h. **P < 0.01 for LPS + CFTRinh-172 vs LPS only. d–f Production of MIP-2 (d) and interleukin-10 (e) and NF-κB activation (f) in the cultured bone-marrow derived wild-type neutrophils and F508del neutrophils simulated with LPS at 12 h. *P < 0.05, **P < 0.01 for wild-type neutrophils receiving LPS vs F508del neutrophils stimulated with LPS. N = 6–8 in each group. Values are mean ± SD
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Fig4: Pro- and anti-inflammatory cytokines in neutrophils balance is impaired without functional CFTR. a–c Production of MIP-2 (a) and interleukin-10 (b) and NF-κB activation (c) in the cultured bone-marrow derived wild-type neutrophils pretreated with CFTRinh-172 and then stimulated with LPS at 12 h. **P < 0.01 for LPS + CFTRinh-172 vs LPS only. d–f Production of MIP-2 (d) and interleukin-10 (e) and NF-κB activation (f) in the cultured bone-marrow derived wild-type neutrophils and F508del neutrophils simulated with LPS at 12 h. *P < 0.05, **P < 0.01 for wild-type neutrophils receiving LPS vs F508del neutrophils stimulated with LPS. N = 6–8 in each group. Values are mean ± SD
Mentions: To study whether lack of functional CFTR in neutrophils affects both pro- and anti-inflammatory cytokines, isolated bone marrow neutrophils were pretreated with CFTRinh-172 (5 μM), stimulated with LPS, and cultured for 12 h to measure MIP-2 and IL-10 in the medium. The adherent neutrophils were then harvested for measurement of p65 NF-κB subunit in nuclear extract; MIP-2 levels increased more than two-fold (Fig. 4a) and IL-10 was significantly decreased (Fig. 4b) in the medium. The p65 NF-κB subunit in the neutrophil nucleus extract with LPS plus CFTRinh-172 inhibition was increased two-fold compared to LPS only (Fig. 4c). To confirm this result, F508del and wild-type neutrophils were cultured and stimulated with LPS for 12 h to measure MIP-2 and IL-10 in the medium. The cells were then harvested for measurement of p65 NF-κB in the nucleus. MIP-2 levels were elevated three-fold (Fig. 4d) and IL-10 was reduced (Fig. 4e). There was increased p65 NF-κB translocation to the nucleus in the F508del neutrophils stimulated with LPS compared to wild-type neutrophils receiving LPS challenge (Fig. 4f).Fig. 4

Bottom Line: Cystic fibrosis transmembrane conductance regulator (CFTR) regulates infection and inflammation.Wild-type mice were reconstituted with F508del neutrophils or bone marrow and then intratracheally challenged with LPS to observe lung inflammatory response.Pharmacologic inhibition and genetic mutation of CFTR in neutrophils activated NF-κB and facilitated macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-α (TNF-α) production.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Research Institute, University of California, San Francisco, HSW 825, 505 Parnassus AVE, San Francisco, CA 94143-0130, USA. suxiao1@yahoo.com

ABSTRACT

Objective and design: Cystic fibrosis transmembrane conductance regulator (CFTR) regulates infection and inflammation. In this study, we investigated whether a lack of functional CFTR in neutrophils would promote lipopolysaccharide (LPS)-induced lung inflammation and injury.

Materials and methods: CFTR-inhibited or F508del-CFTR-mutated neutrophils were stimulated with LPS and cultured to evaluate production of cytokines and NF-κB activation. Wild-type mice were reconstituted with F508del neutrophils or bone marrow and then intratracheally challenged with LPS to observe lung inflammatory response.

Results: Pharmacologic inhibition and genetic mutation of CFTR in neutrophils activated NF-κB and facilitated macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-α (TNF-α) production. Wild-type mice reconstituted with F508del neutrophils and bone marrow had more severe lung inflammation and injury after LPS challenge compared to wild-type mice receiving wild-type neutrophils or bone marrow reconstitution.

Conclusions: Lack of functional CFTR in neutrophils can promote LPS-induced acute lung inflammation and injury.

Show MeSH
Related in: MedlinePlus