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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

Pharmacologic inhibition or mutation of CFTR neutrophils affects proinflammatory cytokine production and NF-κB activation. a, b Cytokine levels in the supernatant of cultured neutrophils at 4 h. a TNF-α. b MIP-2. *P < 0.05 for LPS vs LPS + MalH-2; **P < 0.01 vs PBS or MalH-2 only. c MIP-2 levels in the supernatant of cultured neutrophils stimulated with LPS at 12 h. **P < 0.01 for wild-type neutrophils receiving MalH-2 or F508del neutrophils stimulated with LPS vs LPS only. d Effect of inhibition of NF-κB on production of MIP-2 in wild-type neutrophils with CFTR inhibition by MalH-2. **P < 0.01 for LPS vs LPS + MalH-2; ##P < 0.01 for LPS vs LPS MalH-2 + BMS-34553. N = 6–8 in each group. Values are mean ± SD
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Fig3: Pharmacologic inhibition or mutation of CFTR neutrophils affects proinflammatory cytokine production and NF-κB activation. a, b Cytokine levels in the supernatant of cultured neutrophils at 4 h. a TNF-α. b MIP-2. *P < 0.05 for LPS vs LPS + MalH-2; **P < 0.01 vs PBS or MalH-2 only. c MIP-2 levels in the supernatant of cultured neutrophils stimulated with LPS at 12 h. **P < 0.01 for wild-type neutrophils receiving MalH-2 or F508del neutrophils stimulated with LPS vs LPS only. d Effect of inhibition of NF-κB on production of MIP-2 in wild-type neutrophils with CFTR inhibition by MalH-2. **P < 0.01 for LPS vs LPS + MalH-2; ##P < 0.01 for LPS vs LPS MalH-2 + BMS-34553. N = 6–8 in each group. Values are mean ± SD

Mentions: To test whether inhibition of CFTR in neutrophils would increase proinflammatory cytokine production, we cultured isolated wild-type bone marrow neutrophils for 4 h and measured two important pro-inflammatory cytokines/chemokines—TNF-α and MIP-2—in the medium supernatant. In LPS-stimulated bone marrow neutrophils, both TNF-α and MIP-2 production were increased, and inhibition of CFTR with MalH-2 further increased TNF-α and MIP-2 production (Fig. 3a, b). At 12 h, MalH-2 significantly increased MIP-2 production in LPS-stimulated wild-type neutrophils compared to LPS only (Fig. 3c). For confirmation, we used F508del neutrophils as a positive control. At 12 h, MIP-2 levels in F508del neutrophils stimulated with LPS were increased by three-fold compared to the wild-type neutrophils stimulated with LPS (Fig. 3c), suggesting that CFTR is a modulator of proinflammatory cytokines in bone marrow neutrophils. To investigate whether CFTR inhibition induced excessive MIP-2 production in neutrophils through an NF-κB pathway, LPS simulated wild-type neutrophils were pretreated either with BMS-345541 (50 μM, which was chosen to reach maximal inhibitory effects on NF-κB in neutrophils) or BMS-345541 + MalH-2. BMS-345541 pretreated neutrophils stimulated with LPS produced less MIP-2 compared to LPS alone. BMS-345541 also completely inhibited excessive MIP-2 production in the LPS-stimulated neutrophils with MalH-2 pretreatment (Fig. 3d), suggesting that CFTR regulates MIP-2 production through the NF-κB pathway in LPS-stimulated bone marrow neutrophils.Fig. 3


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)

Pharmacologic inhibition or mutation of CFTR neutrophils affects proinflammatory cytokine production and NF-κB activation. a, b Cytokine levels in the supernatant of cultured neutrophils at 4 h. a TNF-α. b MIP-2. *P < 0.05 for LPS vs LPS + MalH-2; **P < 0.01 vs PBS or MalH-2 only. c MIP-2 levels in the supernatant of cultured neutrophils stimulated with LPS at 12 h. **P < 0.01 for wild-type neutrophils receiving MalH-2 or F508del neutrophils stimulated with LPS vs LPS only. d Effect of inhibition of NF-κB on production of MIP-2 in wild-type neutrophils with CFTR inhibition by MalH-2. **P < 0.01 for LPS vs LPS + MalH-2; ##P < 0.01 for LPS vs LPS MalH-2 + BMS-34553. N = 6–8 in each group. Values are mean ± SD
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Fig3: Pharmacologic inhibition or mutation of CFTR neutrophils affects proinflammatory cytokine production and NF-κB activation. a, b Cytokine levels in the supernatant of cultured neutrophils at 4 h. a TNF-α. b MIP-2. *P < 0.05 for LPS vs LPS + MalH-2; **P < 0.01 vs PBS or MalH-2 only. c MIP-2 levels in the supernatant of cultured neutrophils stimulated with LPS at 12 h. **P < 0.01 for wild-type neutrophils receiving MalH-2 or F508del neutrophils stimulated with LPS vs LPS only. d Effect of inhibition of NF-κB on production of MIP-2 in wild-type neutrophils with CFTR inhibition by MalH-2. **P < 0.01 for LPS vs LPS + MalH-2; ##P < 0.01 for LPS vs LPS MalH-2 + BMS-34553. N = 6–8 in each group. Values are mean ± SD
Mentions: To test whether inhibition of CFTR in neutrophils would increase proinflammatory cytokine production, we cultured isolated wild-type bone marrow neutrophils for 4 h and measured two important pro-inflammatory cytokines/chemokines—TNF-α and MIP-2—in the medium supernatant. In LPS-stimulated bone marrow neutrophils, both TNF-α and MIP-2 production were increased, and inhibition of CFTR with MalH-2 further increased TNF-α and MIP-2 production (Fig. 3a, b). At 12 h, MalH-2 significantly increased MIP-2 production in LPS-stimulated wild-type neutrophils compared to LPS only (Fig. 3c). For confirmation, we used F508del neutrophils as a positive control. At 12 h, MIP-2 levels in F508del neutrophils stimulated with LPS were increased by three-fold compared to the wild-type neutrophils stimulated with LPS (Fig. 3c), suggesting that CFTR is a modulator of proinflammatory cytokines in bone marrow neutrophils. To investigate whether CFTR inhibition induced excessive MIP-2 production in neutrophils through an NF-κB pathway, LPS simulated wild-type neutrophils were pretreated either with BMS-345541 (50 μM, which was chosen to reach maximal inhibitory effects on NF-κB in neutrophils) or BMS-345541 + MalH-2. BMS-345541 pretreated neutrophils stimulated with LPS produced less MIP-2 compared to LPS alone. BMS-345541 also completely inhibited excessive MIP-2 production in the LPS-stimulated neutrophils with MalH-2 pretreatment (Fig. 3d), suggesting that CFTR regulates MIP-2 production through the NF-κB pathway in LPS-stimulated bone marrow neutrophils.Fig. 3

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