Alveolar macrophages play an important role in chronic obstructive pulmonary disease via production of matrix metalloproteinases (MMPs) and cathepsins as well as their inhibitors, tissue inhibitors of metalloproteinases and cystatin C. We hypothesised that expression levels of these molecules by alveolar macrophages at baseline and after stimulation would be influenced by genotype and associated with chronic obstructive pulmonary disease phenotypes. Quantitative PCR and ELISAs/gelatine zymography were used to investigate expression levels of mRNA and protein, respectively. The relationships of expression with genotype, pulmonary function and emphysema were analysed. The results showed that basal expression level of MMP12 mRNA was inversely related to the diffusing capacity of the lung for carbon monoxide/alveolar volume and to forced expiratory volume in 1 s/forced vital capacity after correction for multiple comparisons. The expression level of MMP12 protein stimulated with lipopolysaccharide was also inversely related to the diffusing capacity of the lung for carbon monoxide/alveolar volume and was positively related to the extent of emphysema. The basal expression of MMP1 mRNA was positively correlated with the extent of emphysema. Cathepsin L protein level was positively associated with forced expiratory volume in 1 s % predicted. We conclude that increased MMP12 and MMP1 expression may play a role in the pathogenesis of emphysema. Cathepsin L and MMP9 may be involved in the development of airflow limitation.

Background An imbalance between proteolytic enzymes and their inhibitors is thought to be involved in the pathogenesis of chronic obstructive pulmonary disease. Matrix metalloproteinase-1, also known as interstitial collagenase, has been implicated as a potentially important proteinase in the genesis of chronic obstructive pulmonary disease and, more specifically, emphysema. Methods We performed quantitative immunohistochemical assessment of matrix metalloproteinase-1 expression in the resected lung of 20 smokers/ex-smokers who had varying severity of airflow obstruction and emphysema and compared this with the lungs of 5 nonsmokers. Emphysema was measured using a morphometric measure of the lungs’ surface area/volume ratio and with qualitative and quantitative computed tomography (CT) measures of emphysema. Results There were significantly more matrix metalloproteinase-1-expressing alveolar macrophages and type II pneumocytes as well as a greater percentage of small airways that stained positively for matrix metalloproteinase-1 in the lungs of smokers than in those of nonsmokers ( p < 0.0001, p < 0.0001, and p = 0.0003, respectively). The extent of staining of type II pneumocytes and airways for matrix metalloproteinase-1 was significantly related to the extent of smoking ( p = 0.012 and p = 0.013, respectively). In addition, the extent of matrix metalloproteinase-1 staining of alveolar macrophages was related to the lung surface area/volume ratio and to qualitative estimates of emphysema on CT. Conclusion These findings suggest that cigarette smoking increases expression of matrix metalloproteinase-1 in alveolar macrophages as well as in alveolar and small airway epithelial cells. Smokers who develop emphysema have increased alveolar macrophage expression of matrix metalloproteinase-1.

Palovarotene is an oral γ-selective retinoid agonist. In animal emphysema models, palovarotene reduced inflammation, promoted structural repair and functional improvement.REPAIR (Retinoid treatment of Emphysema in Patients on the α1-antitrypsin International Registry), was an investigator-initiated, double-blind, placebo-controlled randomised study to assess the safety and efficacy of 5 mg·day−1palovarotene given for 1 year to 262 patients with severe α1-antitrypsin deficiency and emphysema confirmed by computed tomography. Change in volume-adjusted 15th percentile point lung density from baseline in 1 year was the primary end-point; functional end-points were also regularly assessed.We randomly assigned 133 and 129 patients to placebo or palovarotene, respectively. Both groups were well matched for all baseline characteristics, including respiratory medications. 88% and 85% of patients completed 1 year of treatment with placebo and palovarotene, respectively. Palovarotene was generally well tolerated. In the study completers population, the placebo-corrected difference of lung density was -0.45 HU at week 28 (p=0.64) and -0.25 HU at week 52 (p=0.94). A nonsignificant treatment difference in most functional parameters of the lung in favour of the drug was observed over time suggesting potential pharmacological effects of palovarotene.Palovarotene 5 mg·day−1over 1 yr failed to show a significant benefit on lung density in moderate-to-severe emphysema secondary to severe α1-antitrypsin deficiency.

Aims: Alpha(1)-antitrypsin deficiency (AATD) is a clinically under-diagnosed genetic disorder that originates from deleterious mutations in the alpha(1)-antitrypsin (AAT) gene, SERPINA1. Severe deficiency is associated with significant pulmonary and hepatic malfunctions. Conventional clinical diagnosis involves the evaluation of serum AAT level and detection of diseased protein isoforms. In this communication, we describe the investigations of a case of severe AATD in which the AAT levels were well below those expected from the MZ phenotype determined by isoelectric focusing for protease inhibitor type (IEF PI-typing).Methods: In addition to the traditional diagnostic method that combines the assessment of serum MT concentration and IEF PI-typing, we investigated the SERPINA1 gene of the proband and participating family members for mutations using Sanger sequencing.Results: We identified a novel mutation (M409T) in the proband, initially missed by the standard diagnostic approach. The novel mutation was present in 4 out of 8 family members who participated in the study.Conclusions: This report illustrates the diagnostic value of incorporating exon sequencing of the MT gene into the algorithm for evaluating AATD, particularly when the AAT serum level is significantly lower than expected from IEF PI-typing. (C) 2012 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Severe α1-antitrypsin deficiency (AATD) is an inherited disorder, leading to development of emphysema in smokers at a relatively young age with disability in their forties or fifties. The emphysema results from excessive elastin degradation by neutrophil elastase as a result of the severe deficiency of its major inhibitor α1-antitrypsin (AAT). The AAT expression is determined by the SERPINA1 gene which expresses codominant alleles. The three most common alleles are the normal M, the S with plasma levels of 60% of normal, and the severely deficient Z with levels of about 15% of normal. Homozygosity for the Z mutant allele is associated with retention of abnormal AAT in the liver, which may lead to neonatal hepatitis, liver disease in children, and liver disease in adults. Regular intravenous infusions of purified human AAT (AAT augmentation therapy) have been used to partially correct the biochemical defect and protect the lung against further injury. Two randomized controlled trials showed a trend of slower progression of emphysema by chest computerized tomography. Integrated analysis of these two studies indicated significantly slower progression of emphysema. AAT is quantified by immunologic measurement of AAT in serum, the phenotype characterized by isoelectric focusing, the common genotypes by targeted DNA analysis, and by sequencing the coding region of the gene when the AAT abnormality remains undefined. AATD is often unrecognized, and diagnosis delayed. Testing for AATD is recommended in patients with chronic irreversible airflow obstruction, especially in those with early onset of disease or positive family history. Testing is also recommended for immediate family members of those with AATD, asthmatics with persistent airflow obstruction, and infants and older subjects with unexplained liver disease. There are over 100 different AAT gene variants; most are rare and only some are associated with clinical disease.

OBJECTIVES: The primary objective of this retrospective study was to evaluate whether abnormal predicted postoperative variables and predicted postoperative product are useful in predicting postoperative complications. The secondary objective was to assess whether an abnormal diffusion heterogeneity index is associated with increased postoperative complications. METHODS: In this retrospective study we evaluated the medical records of 57 patients who underwent lung resection for lung cancer. Calculations of the predicted postoperative variables were done using preoperative testing data, including the extent of the resected lung segments. Predicted postoperative product was obtained by multiplying the predicted postoperative percent-of-predicted FEV1 by the predicted postoperative percent-of-predicted single-breath diffusing capacity of the lung for carbon monoxide (DLCO). The measured product was obtained by multiplying FEV1 by DLCO. We derived diffusion heterogeneity index from measurements of the single-breath DLCO with the 3-equation method, as a measure of the heterogeneity of the distribution of gas exchange in the lung. RESULTS: Patients with complications had lower predicted postoperative FEV1 (P < .001), lower predicted postoperative DLCO (P < .001), lower predicted postoperative maximal oxygen uptake (P < .001), lower predicted postoperative increase in percent-of-predicted DLCO at 70% work load from at-rest percent-of-predicted DLCO (ΔDLCO%) (P < .001), lower predicted postoperative product (P < .001), and lower measured product (P = .004). Interestingly, diffusion heterogeneity index increased with exercise in all patients with complications but decreased with exercise in all patients without complications. CONCLUSIONS: The predicted postoperative variables, predicted postoperative product, measured product, and diffusion heterogeneity index are potentially useful predictors of complications in candidates for lung resection.

BACKGROUND:Oxidative stress induced by smoking is considered to be important in the pathogenesis of Chronic Obstructive Pulmonary Disease (COPD). Heme oxygenase-1 (HMOX1) is an essential enzyme in heme catabolism that is induced by oxidative stress and may play a protective role as an antioxidant in the lung. We determined whether HMOX1 polymorphisms were associated with lung function in COPD patients and whether the variants had functional effects.METHODS:We genotyped five single nucleotide polymorphisms (SNPs) in the HMOX1 gene in Caucasians who had the fastest (n = 278) and the slowest (n = 304) decline of FEV1 % predicted, selected from smokers in the NHLBI Lung Health Study. These SNPs were also studied in Caucasians with the lowest (n = 535) or the highest (n = 533) baseline lung function. Reporter genes were constructed containing three HMOX1 promoter polymorphisms and the effect of these polymorphisms on H2O2 and hemin-stimulated gene expression was determined. The effect of the HMOX1 rs2071749 SNP on gene expression in alveolar macrophages was investigated.RESULTS:We found a nominal association (p = 0.015) between one intronic HMOX1 SNP (rs2071749) and lung function decline but this did not survive correction for multiple comparisons. This SNP was in perfect linkage disequilibrium with rs3761439, located in the promoter of HMOX1. We tested rs3761439 and two other putatively functional polymorphisms (rs2071746 and the (GT)n polymorphism) in reporter gene assays but no significant effects on gene expression were found. There was also no effect of rs2071749 on HMOX1 gene expression in alveolar macrophages.CONCLUSIONS:We found no association of the five HMOX1 tag SNPs with lung function decline and no evidence that the three promoter polymorphisms affected the regulation of the HMOX1 gene.

This review covers protease-antiprotease imbalance in the development of emphysema in smokers. This imbalance is likely to play a major pathogenic role in the development of emphysema in subjects with severe alpha1-antitrypsin deficiency who smoke because of a deficient antiprotease protection against neutrophil elastase release in the lung. Neutrophil elastase is a potent elastolytic enzyme, and its instillation in the lungs of animals results in emphysema. Smoking attracts neutrophils to the lungs and there is an additional accumulation of neutrophils, because the abnormal antitrypsin polymerizes in the lungs and acts as a chemo-attractant to neutrophils. In subjects who do not have antitrypsin deficiency, the case for elastolytic injury by neutrophils causing emphysema is less definite, because of the lack of a severe deficiency of active alpha1-ntitrypsin leading to unopposed elastolysis by neutrophil elastase. It is likely that alveolar macrophages play a pathogenic role in emphysema; they express potent elastolytic enzymes, cathepsins and matrix metalloproteases (MMPs), which are induced by smoking. The numbers of macrophages are increased in the region of the respiratory bronchiole, where centrilobular emphysema develops in smokers. Macrophage cathepsins are inhibited by an antiprotease cystatin C, while the MMPs are inhibited by the tissue inhibitors of metalloproteases (TIMPs). Some pro-inflammatory mediators induce release of MMPs from macrophages without inducing increase in TIMPs, leading to possible protease-antiprotease imbalance. Studies of proteases in alveolar macrophages obtained by bronchoalveolar lavage and studies on lung tissue indicate increased protease expression in subjects with chronic obstructive pulmonary disease (COPD) compared to subjects without COPD.

An abnormal increase in proteolytic enzymes is thought to play a key role in pulmonary emphysema. Alveolar macrophage proteolytic enzymes include cathepsin L, cathepsin S, matrix metalloproteinase 1, 9, and 12, and a number of studies have implicated these proteinases in the alveolar destruction that characterizes emphysema. The aim of this study was to investigate cathepsin L, cathepsin S, matrix metalloproteinase 1, 9, and 12 mRNA expression in alveolar macrophages isolated from patients with varying degrees of emphysema and to correlate their level of expression with measures of emphysema. Alveolar macrophages were isolated from fifty-four patients who underwent surgical resection for lung carcinoma. The level of mRNA expression was determined using real-time PCR. Emphysema was quantified using high-resolution CT scans. Alveolar macrophages were also cultured for 24 h and 48 h; the effect of proinflammatory mediators and promoter polymorphisms on expression was analyzed. There was a significant correlation between matrix metalloproteinase 1 mRNA expression and emphysema. A higher level of matrix metalloproteinase 1 mRNA was associated with more severe emphysema. Matrix metalloproteinase 12 mRNA expression was increased in current smokers as compared with former smokers. Furthermore, there was a negative correlation between matrix metalloproteinase 12 gene expression and carbon monoxide diffusing capacity. The matrix metalloproteinase 9 C-1562T polymorphism significantly influenced matrix metalloproteinase 9 mRNA expression in alveolar macrophages. These results suggest that alveolar macrophage matrix metalloproteinase 1 and 12 may have a role in the lung structural changes leading to the development of emphysema. Furthermore, these data provide evidence to support the concept that multiple proteinases, causing both elastin and collagen degradation, are important in the pathogenesis of pulmonary emphysema.

Objective: The impact of current employment status and other factors on quality of life was evaluated for workers diagnosed with western red cedar asthma in British Columbia, Canada. Methods: Telephone interviews by questionnaire included the Short Form 36 (SF-36) and Marks Asthma quality-of-life instruments. Results: Of the 302 subjects contacted, 70.5% (n = 213) participated. Employment status was the most consistent predictor of quality-of-life domains, with highest scores for employed subjects, particularly those who were no longer exposed to red cedar. Subjects who had quit work because of their asthma had worse scores, particularly for vitality and general health perceptions. Other factors independently associated with specific aspects of poor quality of life were having asthma-like symptoms, taking medication, and not being married. Conclusions: Continued employment was associated with better quality of life for workers with western red cedar asthma.

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Severe alpha1-antitrypsin (AAT) deficiency is an inherited disorder, leading to the development of emphysema in smokers at a relatively young age and disability in most smokers in their 40s. Panacinar emphysema is caused by damage to lung elastin and connective tissue as a result of smoking-induced release of neutrophil elastase in the lung, inadequately inhibited by the severe deficiency in AAT, the major inhibitor of neutrophil elastase. Severe antitrypsin deficiency (mostly protease inhibitor phenotype ZZ) is associated with retention of abnormal AAT in the liver, which may lead to neonatal hepatitis, liver disease in children, and liver disease and cirrhosis in adults. AAT replacement therapy by intravenous infusions of purified human AAT has been used to partially correct the biochemical defect and protect the lung against further injury, but has not yet been proven to be clinically effective in reducing the progression of disease in AAT-deficient patients. However, there was a suggestion of a slower progression of emphysema by chest computerized tomography in a small randomized trial, and non-randomized observational studies suggested a slower rate of decline in lung function in subjects with moderate airflow obstruction. AAT deficiency is often unrecognized. AAT testing is recommended in patients with chronic irreversible airflow obstruction with early onset of disease, those with disability in their 40s or 50s or positive family history, in immediate family members of AAT-deficient patients, in infants and children with liver disease, and adults with unexplained liver disease.

OBJECTIVE To evaluate the effect of lung resection on lung function and exercise capacity values, including diffusion capacity of the lung for carbon monoxide (Dlco), during exercise, and to determine whether postoperative lung function, including exercise capacity and Dlco during exercise, could be predicted from preoperative lung function and the number of functional segments resected. DESIGN Prospective study. SETTING Clinical pulmonary function laboratory in a university teaching hospital. PATIENTS Twenty-eight patients undergoing lung resection at Vancouver General Hospital from October 1998 to May 1999, were studied preoperatively and 1-year postoperatively. INTERVENTIONS We determined FEV(1) and FVC, and maximal oxygen uptake (Vo(2)max) and maximal workload (Wmax) achieved during incremental exercise testing. We used the three-equation modification of the single-breath Dlco technique to determine Dlco at rest (RDlco) and during steady-state exercise at 70% of Wmax, and the increase in Dlco from rest to exercise (ie, the mean increase in Dlco percent predicted at 70% of Wmax from resting Dlco percent predicted [(70%-R)Dlco]). We calculated the predicted postoperative (PPO) values for all the above parameters using the preoperative test data and the extent of functioning bronchopulmonary segments resected, and compared the results with the actual 1-year postoperative results. RESULTS Following lung resection, there was a significant reduction in FEV(1), FVC, and Dlco with decreases of 12%, 13%, and 22% predicted, respectively. There were also significant decreases in Vo(2)max per kilogram of 2.1 mL/min/kg (8% of predicted Vo(2)max) and in Wmax of 12 W (7% of predicted Wmax). However, (70%-R)Dlco did not significantly decrease after lobectomy but decreased after pneumonectomy. The calculated PPO values significantly underestimated postoperative values after pneumonectomy but were acceptable for lobectomy. CONCLUSIONS Exercise tests may be better indicators of functional capacity after lung resection than measurements of FEV(1) and FVC or RDlco. PPO results calculated by estimating the functional contribution of the resected segments, are comparable with those obtained using ventilation-perfusion lung scanning and significantly underestimate postoperative lung function after pneumonectomy, but are acceptable for lobectomy.

Pulmonary emphysema is a major cause of mortality and morbidity in chronic obstructive pulmonary disease (COPD). Cigarette smoking is a major risk factor in the development of pulmonary emphysema. In this study, we investigated the acute effect of cigarette smoke in vitro on the production of tumour necrosis factor-α (TNF-α) using differentiated U937 cells, a macrophage model system. We found that stimulation of the macrophages with cigarette smoke media (CSM) leads to a rapid activation of extracellular-regulated kinases 1 and 2 (erk1/2), p90RSK and a transient decrease in phosphorylation of PKB/akt. The CSM also caused the subsequent induction of TNF-α release. Our studies revealed that U0126, an inhibitor of the erk1/2 pathway, markedly suppressed CSM-induced TNF-α release. Consistent with this finding, U0126 blocked CSM-stimulated erk1/2 phosphorylation, as well as phosphorylation of the downstream kinase, p90RSK. On the other hand, the PI3-K inhibitor, LY294002, and epidermal growth factor receptor (EGFR)-specific inhibitor, AG1478, did not suppress the release of TNF-α. Thus, CSM induction of TNF-α production by differentiated macrophages is regulated primarily via the erk1/2 pathway.