Limits...
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.

Show MeSH

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
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3116128&req=5

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
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3116128&req=5

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.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