Inhibition of IP-10 secretion in lung epithelial cell/PBMC co-cultures by human IFN-γ antibody

<p><b>Copyright information:</b></p><p>Taken from "The role of IFN-γ in regulation of IFN-γ-inducible protein 10 (IP-10) expression in lung epithelial cell and peripheral blood mononuclear cell co-cultures"</p><p>http://respiratory-research.com/content/8/1/80</p><p>Respiratory Research 2007;8(1):80-80.</p><p>Published online 8 Nov 2007</p><p>PMCID:PMC2174934.</p><p></p

IL-12 (100 ng/ml) mediated secretion of IP-10 in lung epithelial cell/PBMC co-cultures

<p><b>Copyright information:</b></p><p>Taken from "The role of IFN-γ in regulation of IFN-γ-inducible protein 10 (IP-10) expression in lung epithelial cell and peripheral blood mononuclear cell co-cultures"</p><p>http://respiratory-research.com/content/8/1/80</p><p>Respiratory Research 2007;8(1):80-80.</p><p>Published online 8 Nov 2007</p><p>PMCID:PMC2174934.</p><p></p> Data represent the mean ± SEM of 7–14 independent experiments, **p < 0.01, ANOVA with Bonferroni's multiple comparison test

Basal, IFN-γ and IL-12 mediated secretion of IP-10 in lung epithelial cell/PBMC co-cultures cultured in transwell chambers with separating filters

<p><b>Copyright information:</b></p><p>Taken from "The role of IFN-γ in regulation of IFN-γ-inducible protein 10 (IP-10) expression in lung epithelial cell and peripheral blood mononuclear cell co-cultures"</p><p>http://respiratory-research.com/content/8/1/80</p><p>Respiratory Research 2007;8(1):80-80.</p><p>Published online 8 Nov 2007</p><p>PMCID:PMC2174934.</p><p></p> Data represent the mean ± SEM of 4 independent experiments. Control (□) values are shown as IP-10 secretion in lung epithelial cell/PBMC co-cultures cultured in transwell chambers without a separating filter

The dose dependent inhibition of IP-10 secretion by PI3 kinase inhibitor in Calu-3/PBMCs (A

<p><b>Copyright information:</b></p><p>Taken from "The role of IFN-γ in regulation of IFN-γ-inducible protein 10 (IP-10) expression in lung epithelial cell and peripheral blood mononuclear cell co-cultures"</p><p>http://respiratory-research.com/content/8/1/80</p><p>Respiratory Research 2007;8(1):80-80.</p><p>Published online 8 Nov 2007</p><p>PMCID:PMC2174934.</p><p></p>) and A549/PBMCs (B.) co-cultures with or without IFN-γ and IL-12 treatment. The dose dependent inhibition by PI3 kinase inhibitor of IL-12 mediated IFN-γ secretion in A549/PBMCs co-cultures is seen in (C.) Data represent the mean ± SEM of 4–7 independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001, ANOVA with Bonferroni's multiple comparison test

The effects of conditioned media (CM) on IP-10 secretion from cells cultured alone

<p><b>Copyright information:</b></p><p>Taken from "The role of IFN-γ in regulation of IFN-γ-inducible protein 10 (IP-10) expression in lung epithelial cell and peripheral blood mononuclear cell co-cultures"</p><p>http://respiratory-research.com/content/8/1/80</p><p>Respiratory Research 2007;8(1):80-80.</p><p>Published online 8 Nov 2007</p><p>PMCID:PMC2174934.</p><p></p> The lung epithelial cells or PBMCs were cultured for 18 hours in CM simultaneously with either IFN-γ (10 ng/ml) or IL-12 (100 ng/ml) incubation. PBMCs cultured alone were used as control. Data represent the mean ± SEM of 4–6 independent experiments, *p < 0.05, **p < 0.01, ANOVA with Bonferroni's multiple comparison test

Cigarette Smoke Induced Airway Inflammation Is Independent of NF-κB Signalling

<div><h3>Rationale</h3><p>COPD is an inflammatory lung disease largely associated with exposure to cigarette smoke (CS). The mechanism by which CS leads to the pathogenesis of COPD is currently unclear; it is known however that many of the inflammatory mediators present in the COPD lung can be produced via the actions of the transcription factor Nuclear Factor-kappaB (NF-κB) and its upstream signalling kinase, Inhibitor of κB kinase-2 (IKK-2). Therefore the NF-κB/IKK-2 signalling pathway may represent a therapeutic target to attenuate the inflammation associated with COPD.</p> <h3>Aim</h3><p>To use a range of assays, genetically modified animals and pharmacological tools to determine the role of NF-κB in CS-induced airway inflammation.</p> <h3>Methods</h3><p>NF-κB pathway activation was measured in pre-clinical models of CS-induced airway inflammation and in human lung tissue from COPD patients. This data was complemented by employing mice missing a functional NF-κB pathway in specific cell types (epithelial and myeloid cells) and with systemic inhibitors of IKK-2.</p> <h3>Results</h3><p>We showed in an airway inflammation model known to be NF-κB-dependent that the NF-κB pathway activity assays and modulators were functional in the mouse lung. Then, using the same methods, we demonstrated that the NF-κB pathway appears not to play an important role in the inflammation observed after exposure to CS. Furthermore, assaying human lung tissue revealed that in the clinical samples there was also no increase in NF-κB pathway activation in the COPD lung, suggesting that our pre-clinical data is translational to human disease.</p> <h3>Conclusions</h3><p>In this study we present compelling evidence that the IKK-2/NF-κB signalling pathway does not play a prominent role in the inflammatory response to CS exposure and that this pathway may not be important in COPD pathogenesis.</p> </div

Temporal characterisation of NF-κB pathway activation after 3 days of CS challenge.

<p>Mice were challenged for 3 days with CS (500 ml/min, 1 hour, twice daily) or ambient air. Samples were collected at increasing times after the last challenge. NF-κB subunits were measured in the lung nuclear fractions. A: p65, B: p50, C RelB and D: p52. Data are presented as mean ± s.e.m. of n = 6–8 observations.</p

Temporal characterisation of airway inflammation after 14 days of CS challenge.

<p>Mice were challenged for 14 days with CS (500 ml/min, 1 hour, twice daily) or ambient air. Samples were collected at increasing time points after the final challenge. A) BALF neutrophilia, B) BALF macrophage number, C) BALF lymphocytes and D) NF-κB (p65):DNA association in lung nuclear extract. Data are presented as mean ± s.e.m. of n = 6–8 observations.</p

Temporal characterisation of the airway inflammation after LPS or 3 days of CS challenge.

<p>Mice were challenged with LPS (1 mg/ml) or endotoxin free saline for 30 minutes. Samples were collected at increasing time points after challenge. A) LPS-induced BALF neutrophilia. B) LPS-induced NF-κB(p65):DNA association in lung nuclear extract. Mice were challenged for 3 days with CS (500 ml/min, 1 hour, twice daily) or ambient air. Samples were collected at increasing time points after the final challenge. C) CS-induced BALF neutrophilia. D) CS-induced NF-κB (p65):DNA association in lung nuclear extract. Data are presented as mean ± s.e.m. of n = 6–8 observations.</p

Profile of IKK-2^ΔEpi mice in the 3 and 14 day CS-driven models.

<p>IKK-2^ΔEpi mice were challenged with 3 or 14 days CS (500 ml/min, 1 hour, twice daily) or ambient air. BALF samples were collected twenty-four hours after the final challenge. BALF neutrophilia after 3 days of CS challenge (A) BALF neutrophilia, macrophages and lymphocytes after 14 days of CS challenge (B, C and D, respectively). Data are presented as mean ± s.e.m. of n = 8–16 observations. # indicates a statistically significant difference (p<0.05) from air challenged control groups (Mann-Whitney test). Any changes in data collected from the GM control mice i.e. fl/fl, TET alone and CRE alone did not reach statistical significance when compared to the appropriate control group.</p