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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.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.

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.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.

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