elastin and alpha-1-Antitrypsin-Deficiency

Biomarkers of pathogenesis in chronic obstructive pulmonary disease (COPD) can significantly accelerate drug development. In COPD related to alpha-1 antitrypsin deficiency, the role of neutrophil elastase and its inhibition by alpha-1 antitrypsin protein focused interest on elastin degradation and the development of pulmonary emphysema. Amino acids desmosine and isodesmosine are unique cross-links in mature elastin fibers and can serve as biomarkers of elastin degradation when measured in body fluids. This review gives a perspective on what has been learned by the earliest measurements of desmosine and isodesmosine followed by later studies using methods of increased sensitivity and specificity and the meaning for developing new therapies. Also included are brief statements on the biomarkers fibrinogen, CC-16, and Aa-Val-360 in COPD.

Topics: alpha 1-Antitrypsin Deficiency; Biomarkers; Desmosine; Elastin; Fibrinogen; Humans; Isodesmosine; Leukocyte Elastase; Pulmonary Disease, Chronic Obstructive; Pulmonary Emphysema; Uteroglobin

The serine proteinase elastase is located in the azurophil granules of mature circulating polymorphonuclear neutrophils. This neutrophil elastase or NE is a potent non specific serine protease which plays a role as bactericidal agent and in the degradation of immune complexes by intraphagosomal processes. It promotes inflammation when the granule contents are secreted in the extracellular environment. In certain pathological circumstances, an imbalance between NE and its major plasmatic inhibitor alpha 1-PI (formerly, alpha 1-antitrypsin) leads to abnormal tissue destruction and disease development. Genetic or acquired alpha 1-PI deficiency is thought to be involved in the pathogenesis of pulmonary emphysema. A variety of degenerative and degradative disorders are also associated to uncontrolled proteolysis by NE (rheumatoid arthritis, glomerulonephritis, adult respiratory distress symptom, psoriasis, cancer). Numerous inhibitors of NE have been reported. Various molecules are currently undergoing clinical trials for emphysema and other pulmonary diseases.

Topics: alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Animals; Coumarins; Cytoplasmic Granules; Drug Design; Elastin; Genetic Predisposition to Disease; Humans; Leukocyte Elastase; Molecular Structure; Neutrophils; Pancreas; Pancreatic Elastase; Phagosomes; Pulmonary Emphysema; Serine Proteinase Inhibitors; Structure-Activity Relationship; Substrate Specificity

Elastases are proteinases capable of solubilizing fibrous elastin. They may belong to the class of serine proteinases, cysteine proteinases and metalloproteinases. Mammalian elastases occur mainly in the pancreas and the phagocytes. Among non-mammalian elastases there is a great variety of bacterial metallo and serine elastases. The elastolytic activity varies from one elastase to another and is usually not correlated with the catalytic efficiency of these proteinases. One may measure this activity using native or labelled elastins. With pure elastases one may use synthetic substrates. There is a large number of natural (proteins) and synthetic elastase inhibitors. Elastases play a pathologic role in pulmonary emphysema, cystic fibrosis, infections, inflammation and atherosclerosis.

Topics: alpha 1-Antitrypsin Deficiency; alpha-Macroglobulins; Animals; Arteriosclerosis; Arthritis, Rheumatoid; Bacterial Proteins; Catalysis; Cathepsin G; Cathepsins; Elastin; Enzyme Inhibitors; Fibroblasts; Glycosaminoglycans; Humans; Leukocytes; Mammals; Organ Specificity; Pancreas; Pancreatic Elastase; Phagocytes; Polynucleotides; Pseudomonas Infections; Pulmonary Emphysema; Serine Endopeptidases; Species Specificity; Substrate Specificity

Topics: alpha 1-Antitrypsin Deficiency; Animals; Elastin; Humans; Lung; Pancreatic Elastase; Phenotype; Pulmonary Emphysema; Smoking

Topics: Aerosols; alpha 1-Antitrypsin Deficiency; Animals; Blood Platelets; Cadmium; Collagen; Disease Models, Animal; Elastin; Emphysema; Forecasting; Hexanes; Lathyrism; Macrophages; Mice; Mice, Mutant Strains; Microscopy, Electron; Neutrophils; Nutrition Disorders; Oxygen; Pancreatic Elastase; Papain; Physical Exertion; Pulmonary Fibrosis; Pulmonary Gas Exchange; Pulmonary Heart Disease; Respiration; Trypsin

Several inherited syndromes characterized by abnormal elastic fibers decreased in number and size could be collected under the heading of inherited elastolysis. This morphological concept does not prejudge the causal mechanisms of the elastolysis involving dermis and/or other organs. The elastic fibers anomalies result mainly from elastin crosslinking defects, developmental disturbances or excessive proteolysis.

Topics: alpha 1-Antitrypsin Deficiency; Ehlers-Danlos Syndrome; Elastin; Homocystinuria; Humans; Marfan Syndrome; Menkes Kinky Hair Syndrome; Skin; Skin Diseases

Topics: alpha 1-Antitrypsin Deficiency; alpha-Macroglobulins; Animals; Blood Proteins; Cricetinae; Elastin; Humans; In Vitro Techniques; Lung; Macrophages; Neutrophils; Pancreas; Pancreatic Elastase; Pulmonary Emphysema; Smoking

Topics: Adult; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Elastin; Heterozygote; Homozygote; Humans; Middle Aged; Pancreatic Elastase; Phenotype; Protease Inhibitors; Pulmonary Emphysema; Risk; Smoking

Topics: alpha 1-Antitrypsin Deficiency; Amino Acid Chloromethyl Ketones; Elastin; Enzyme Inhibitors; Humans; Pancreatic Elastase; Pulmonary Alveoli; Pulmonary Emphysema

Topics: Adult; Age Factors; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Alpha-Globulins; Animals; Antigen-Antibody Reactions; Cadmium; Collagen; Elastin; Environmental Exposure; Humans; Infant; Leukocytes; Lung; Lung Diseases; Macrophages; Microbial Collagenase; Nitrogen Dioxide; Ozone; Pancreatic Elastase; Peptide Hydrolases; Pulmonary Alveoli; Pulmonary Edema; Pulmonary Emphysema; Smoking

This article assesses developments in cardiorespiratory medicine since the Nobel Prize in Physiology or Medicine was awarded in 1956 for advancements in the study of cardiorespiratory disease. In chronic obstructive pulmonary disease, advances were accelerated by the discovery of a genetically determined cause for pulmonary emphysema in the genetic abnormality alpha-1 antitrypsin deficiency. This causes a deficiency of the inhibitor of neutrophil elastase, which results in increased degradation of lung elastin and the development of pulmonary emphysema. This discovery gave focus to two amino acids that reside only in body elastin, desmosine and isodesmosine, which can be measured as biomarkers of elastin degradation in body fluids with increased accuracy and sensitivity. Studies of this biomarker have shown that augmentation therapy in alpha-1 antitrypsin deficiency does decrease lung and body elastic tissue degradation and in the RAPID (Randomized, Placebo-controlled Trial of Augmentation Therapy in Alpha-1 Proteinase Inhibitor Deficiency) Study, over 4 years, showed a preservation of lung density by computer tomography correlating with decreases in plasma levels of desmosine and isodesmosine. This insight indicates the potential of agents that prevent lung elastin degradation. Such an agent is hyaluronan aerosol, which is deficient in post mortem lungs with chronic obstructive pulmonary disease and has been shown to block elastin degradation, possibly by a barrier function. Thus it would appear that hyaluronan could have therapeutic potential in chronic obstructive pulmonary disease.

Topics: alpha 1-Antitrypsin Deficiency; Animals; Biomarkers; Desmosine; Elastin; Humans; Isodesmosine; Leukocyte Elastase; Pulmonary Disease, Chronic Obstructive; Pulmonary Emphysema

Intravenous alpha-1 antitrypsin protein (AAT) augmentation is a prescribed therapy for severe, genetically determined, alpha-1 antitrypsin deficiency (AATD), a genetic basis for pulmonary emphysema. AAT, a predominant systemic inhibitor of neutrophil elastase thus far has not been shown to decrease elastin degradation in a significant number of patients on this therapy. The objective of this study was to compare levels of biomarkers of elastin degradation in plasma, bronchoalveolar lavage (BALF) fluid and urine before and after beginning AAT augmentation therapy in patients with AATD.. Desmosine and isodesmosine (DI), which occur only in elastin, are amino acid cross-links in mature elastin. Levels of DI in body fluids measure degradation of elastin and can be measured more specifically by mass spectrometry. This method was used to measure DI levels in plasma, bronchoalveolar lavage fluid and urine in cohorts of severe AATD patients on augmentation, not on augmentation and before and after the initiation of augmentation therapy.. Statistically significant reductions in plasma DI and in BALF DI were demonstrated in AATD patients receiving intravenous (IV) augmentation therapy as compared with those not receiving it. Administration by aerosol also produced statistically significant reductions in levels of DI in BALF.. Results indicate that the currently prescribed doses of AAT augmentation inhibit neutrophil elastase adequately to reduce elastin degradation, both systemically and in the lung per se. The currently prescribed doses did not reduce elastin degradation to control levels, which may be possible with higher doses.

Topics: Aged; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Biomarkers; Bronchoalveolar Lavage Fluid; Elastin; Female; Homozygote; Humans; Isodesmosine; Leukocyte Elastase; Male; Middle Aged; Phenotype; Trypsin Inhibitors

Topics: alpha 1-Antitrypsin Deficiency; Biomarkers; Elastin; Humans; Isodesmosine; Pulmonary Emphysema

Liberation of elastin peptides from damaged lung may be a mechanism of autoimmune lung disease. Citrullination, and anti-citrullinated protein antibody formation occurs in smokers, but the role of smoking in autoantibody generation relevant to pulmonary disease is unclear. Anti-elastin, anti-cyclic citrullinated peptide (anti-CCP) and anti-mutated citrullinated vimentin (anti-MCV) antibodies were measured in 257 subjects with α₁-antitrypsin deficiency (AATD), 113 subjects with usual chronic obstructive pulmonary disease (COPD) and 22 healthy nonsmokers. Levels were compared between groups, against phenotypic features and against smoke exposure. Anti-elastin antibodies were higher in controls relative to AATD (p = 0.008) and usual COPD (p < 0.00001), and in AATD relative to usual COPD (p < 0.00001). Anti-elastin levels showed a threshold at 10 pack-yrs, being higher in those who had smoked less (p = 0.004). No relationships between antibody levels and clinical phenotype were seen after adjustment for smoke exposure. Anti-CCP antibodies were higher in usual COPD than AATD (p = 0.002) but the relationship to smoke exposure was less clear. Smoke exposure is the main determinant of anti-elastin antibody levels, which fall after 10 pack-yrs. Local antibody complexes may be a better measure of elastin directed autoimmunity than circulating levels.

Topics: Adult; Aged; alpha 1-Antitrypsin Deficiency; Autoimmune Diseases; Elastin; Female; Forced Expiratory Volume; Humans; Lung Diseases; Male; Middle Aged; Phenotype; Pulmonary Disease, Chronic Obstructive; Risk; Smoking; Time Factors

In patients with chronic inflammatory lung disease, pulmonary proteases can generate neoantigens from elastin and collagen with the potential to fuel autoreactive immune responses. Antielastin peptide antibodies have been implicated in the pathogenesis of tobacco-smoke-induced emphysema. Collagen-derived peptides may also play a role.. To determine whether autoantibodies directed against elastin- and collagen-derived peptides are present in plasma from three groups of patients with chronic inflammatory lung disease compared with a nonsmoking healthy control group and to identify whether autoimmune responses to these peptides may be an important component of the disease process in these patients.. A total of 124 patients or healthy control subjects were recruited for the study (Z-A1AT deficiency, n = 20; cystic fibrosis, n = 40; chronic obstructive pulmonary disease, n = 31; healthy control, n = 33). C-reactive protein, IL-32, and antinuclear antibodies were quantified. Antielastin and anti-N-acetylated-proline-glycine-proline autoantibodies were measured by reverse ELISA.. All patients were deemed stable and noninfective on the basis of the absence of clinical or radiographic evidence of recent infection. There were no significant differences in the levels of autoantibodies or IL-32 in the patients groups compared with the healthy control subjects.. Antielastin or anti-N-acetylated proline-glycine-proline autoantibodies are not evident in chronic inflammatory lung disease.

Topics: Adolescent; Adult; Aged; alpha 1-Antitrypsin Deficiency; Autoantibodies; Autoimmune Diseases; Case-Control Studies; Cystic Fibrosis; Elastin; Female; Humans; Male; Middle Aged; Oligopeptides; Pulmonary Disease, Chronic Obstructive; Smoking; Young Adult

Desmosine (DES) and isodesmosine (IDES) are two unusual, tetrafunctional, pyridinium ring-containing amino acids involved in elastin cross-linking. Being amino acids unique to mature, cross-linked elastin, they are useful for discriminating peptides derived from elastin breakdown from precursor elastin peptides. According to these features, DES and IDES have been extensively discussed as potentially attractive indicators of elevated lung elastic fibre turnover and markers of the effectiveness of agents with the potential to reduce elastin breakdown. In the present manuscript, immunology-based and separation methods for the evaluation of DES and IDES are discussed, along with studies reporting increased levels of urine excretion in chronic obstructive pulmonary disease (COPD) patients with and without alpha(1)-antitrypsin deficiency. The results of the application of DES and IDES as surrogate end-points in early clinical trials in COPD are also reported. Finally, recent advances in detection techniques, including liquid chromatography tandem mass spectrometry and high-performance capillary electrophoresis with laser-induced fluorescence, are discussed. These techniques allow detection of DES and IDES at very low concentration in body fluids other than urine, such as plasma or sputum, and will help the understanding of whether DES and IDES are potentially useful in monitoring therapeutic intervention in COPD.

Topics: Adult; alpha 1-Antitrypsin Deficiency; Child; Chromatography, Liquid; Desmosine; Elastin; Female; Humans; Isodesmosine; Male; Models, Biological; Peptides; Pulmonary Disease, Chronic Obstructive; Smoking; Tandem Mass Spectrometry

Application of mass spectrometry (MS) for direct measurements of desmosine (D) and isodesmosine (I) in urine, plasma, and sputum as markers of elastin degradation in patients with alpha(1)-antitrypsin deficiency (AATD) and non-AATD-related COPD.. In COPD patients, the lungs undergo elastin injury, which can be monitored by measurements of D and I in body fluids as specific markers of elastin degradation using the specificity and sensitivity of MS.. Acid hydrolysis of blood plasma, 24-h urine and sputum measurements, followed by chromatographic separation for mass spectrometric analysis.. Each patient group had levels of plasma D and I that were statistically significantly higher than those of control subjects. AATD patients had higher levels than COPD patients with normal alpha(1)-antitrypsin (AAT) levels. Twenty-four-hour urine measurements demonstrated no significant difference in total levels of D and I among control subjects and patients but showed a free (unbound) concentration of D and I in urine, which was statistically significantly higher in patients with COPD with and without AAT. The D and I levels in the sputum of patients with AATD exceeded the levels in COPD patients with normal AAT levels.. MS allows a sensitive and specific analysis of D and I in body fluids. The quantification of D and I in sputum, along with increases of D and I in plasma and an elevated free component of D and I in urine provide indexes that characterize patients with COPD and can be followed in relation to the course of the disease and/or therapy.

Topics: Adult; Aged; Aged, 80 and over; alpha 1-Antitrypsin Deficiency; Desmosine; Elastin; Female; Humans; Isodesmosine; Male; Mass Spectrometry; Matched-Pair Analysis; Middle Aged; Prognosis; Pulmonary Disease, Chronic Obstructive; Sensitivity and Specificity; Sputum

Degradation of extracellular matrix components is central to many pathological features of chronic destructive lung disorders. Desmosine and isodesmosine are elastin-derived cross-linked amino acids whose urine levels are considered representative of elastin breakdown. The aim of this study was to apply a novel methodology, based on high-performance capillary electrophoresis, to the quantification of desmosine and isodesmosine in 11 patients with stable chronic obstructive pulmonary disease (COPD), 10 with an exacerbation of COPD, nine with alpha1-antitrypsin deficiency, 13 with bronchiectasis, and 11 adults with cystic fibrosis, in comparison to 24 controls. It was found that, in patients with stable COPD, urinary desmosine levels were higher than in controls (p=0.03), but lower than in COPD subjects with an exacerbation (p< or =0.05). The highest desmosine levels were found in subjects with alpha1-antitrypsin deficiency, bronchiectasis and cystic fibrosis (p<0.001 versus stable COPD). In a short-term longitudinal study, five stable COPD patients showed a constant rate of desmosine excretion (mean coefficient of variation <8% over three consecutive days). In conclusion, the present method is simple and suitable for the determination of elastin-derived cross-linked amino acid excretion in urine, giving results similar to those obtained using other separation methods. In addition, evidence is presented that urinary desmosine excretion is increased in conditions characterized by airway inflammation, such as exacerbations of chronic obstructive pulmonary disease, bronchiectasis and cystic fibrosis. Results obtained in subjects with alphal-antitrypsin deficiency suggest that this method might be used to evaluate the putative efficacy of replacement therapy.

Topics: Adult; Aged; Aged, 80 and over; alpha 1-Antitrypsin Deficiency; Bronchiectasis; Cross-Linking Reagents; Cross-Sectional Studies; Cystic Fibrosis; Desmosine; Elastin; Electrophoresis, Capillary; Emphysema; Extracellular Matrix; Female; Humans; Isodesmosine; Longitudinal Studies; Lung Diseases, Obstructive; Male; Middle Aged

We evaluated the ability of intravenous supplementation therapy with alpha(1)-antitrypsin (AAT) to reduce the rate of urinary excretion of desmosine (DES), a specific marker of elastin degradation, in eight men and four women with emphysema due to severe, congenital deficiency of AAT (range 17-69 mg/dl). Nine were former cigarette smokers, two were current smokers, and one reported never smoking; their mean age was 54 (SD 12) yr and their mean FEV(1) was 41 (18%) of predicted. Urinary DES was measured by isotope dilution and HPLC. Prior to the start of AAT supplementation, mean DES excretion was 13.0 (5.0) microg/g creatinine, 73% higher than in healthy nonsmokers. During 8 wk of supplementation therapy, mean urinary DES excretion was 13.0 (5.9) microg/g creatinine, unchanged from the baseline period (p = 0.85 by repeated measures ANOVA). We conclude that baseline levels of elastin degradation in emphysematous patients with severe AAT deficiency were abnormally high and that 8 wk of AAT supplementation therapy did not appreciably reduce the rate of elastin degradation. These findings raise the possibilities that protective levels of AAT in the lungs are insufficient or that elastin degradation in the lungs of these subjects is not dependent upon neutrophil elastase at this time.

Topics: Acute Disease; Adult; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Analysis of Variance; Cotinine; Desmosine; Elastin; Female; Humans; Infusions, Intravenous; Lung; Male; Pulmonary Emphysema; Time Factors

Topics: alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Animals; Animals, Newborn; Disease Models, Animal; Elastin; Emphysema; Humans; Leukocyte Elastase; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Mutant Strains; Microscopy, Immunoelectron; Pulmonary Alveoli

The possibility that polymorphonuclear leukocytes (PMN) recruited into the lung have the capability to damage alveolar septa was investigated in several strains of mice with different serum alpha 1 proteinase inhibitor levels and PMN lysosomal functions. After an intratracheal instillation of FMLP (200 micrograms), all strains of mice showed a similar PMN influx in alveolar spaces with an increase (approximately 4- to 5-fold) in bronchoalveolar lavage total cell count, which peaked at 24 to 48 hours. At this time, differential cell count in all strains revealed an approximately 40-fold increase in neutrophils. In C57BL/6J and pallid mice but not in NMRI mice, PMN influx was followed by a decrease in lung elastin content (-17% and -37%, respectively) and by the development of significant emphysema (mean linear intercept, +28% and +56%, respectively). The onset of the pulmonary lesion was preceded by a marked increase of neutrophil elastase burden in alveolar interstitium. Compared with NMRI mice, C57BL/6J and pallid mice have lower serum elastase inhibitory capacity levels. The degree of lung destruction was inversely correlated with elastase inhibitory capacity levels. Lung elastin degradation and emphysema may be induced by eliciting PMN into the lungs only in animals with a deficient anti-elastase screen. Compared with C57BL/6J mice, pallid mice showed a significantly greater lung elastin loss and a higher degree of emphysema after FMLP treatment. These differences may be accounted for by the higher baseline levels of interstitial elastase burden. It may be assumed that an enzymatically active elastase was already working on the lung interstitium before FMLP instillation in pallid mice.

Topics: alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Animals; Bronchoalveolar Lavage Fluid; Chemotaxis, Leukocyte; Elastin; Leukocyte Elastase; Lung; Male; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Neutrophils; Pancreatic Elastase; Pulmonary Emphysema

According to a current hypothesis, pulmonary emphysema results from damage to the elastic fiber network caused by an imbalance within the lower respiratory tract between elastase(s) and protease inhibitors. This hypothesis is based largely on studies of persons with genetic deficiency of serum alpha 1-proteinase inhibitor. We recently reported a spontaneously occurring emphysema in the pallid mouse with an inherited deficiency of serum alpha 1-proteinase inhibitor. This animal reproduces important features of the human condition. We describe here the changes in alveolar elastolytic burden and in the bronchoalveolar lavage (BAL) cell population, which precede and accompany the development of emphysema in pallid mice.. A possible correlation between the levels of lung elastase burden and the loss of lung elastin content was investigated in the period of development of emphysema in pallid mice. Changes in alveolar cells were investigated for specimens from BAL fluids and lung tissue by cytologic, histologic, and immunogold-electron microscopic methods.. An immunogold-positive reaction for elastase was observed on elastin within the alveolar walls of pallid mice from 2 months onward. The average of gold particle density progressively increased with age, reaching high values at 12 and 16 months of age, the age at which parenchymal destruction was previously reported to occur. Lung elastin content had normal values at 2, 4, and 8 months of age. However, it was significantly lower at 12 and 16 months of age. The immunogold values of elastase burden correlated inversely with the decrease in lung elastin content. Total and differential cell counts from BAL fluids of pallid mice did not differ significantly at any time in life and were similar to age-matched controls. However, in pallid mice from 8 months of life onward, pulmonary macrophages contained characteristic intracytoplasmic crystalloid inclusions, which were electrondense and bounded by a single membrane. Using electron microscopy and an immunogold-labeling technique with anti-mouse I-III collagen IgG, these inclusions were identified as collagen-derived products.. The data reported in this paper suggest that emphysematous lesions in pallid mice are associated with a progressive increase of elastase in alveolar intestitium and with loss of lung elastin. Surprisingly, the number of neutrophils in BAL fluids does not increase with the increase of lung elastolytic burden. However, intracytoplasmic crystalloid inclusions related to collagen degradation were observed in pulmonary macrophages of pallid mice at the time of septal disruption. The presence of similar structures in alveolar macrophages from mice or other animal species may be indicative of connective tissue breakdown or remodeling of tissue collagen.

Topics: alpha 1-Antitrypsin Deficiency; Animals; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Elastin; Leukocyte Elastase; Lung; Male; Mice; Mice, Inbred C57BL; Pancreatic Elastase; Pulmonary Emphysema

It is hypothesized that emphysema develops in some severely alpha 1-antitrypsin (AAT)-deficient persons because endogenous elastases are not adequately controlled by AAT, and accelerated elastin degradation occurs. It is not known whether augmentation therapy with AAT diminishes degradation of lung elastin in severely deficient persons with lung disease. Two severely deficient, PiZ patients were studied, a 63-year-old never-smoking woman with bronchiectasis and a 41-year-old smoking man with emphysema. Urinary desmosine (DES) was determined before and after augmentation therapy with AAT, 260 mg/kg/month. Mean +/- SEM pretreatment urinary DES was elevated in both patients, 19.7 +/- 0.9 (n = 2) and 10.8 +/- 0.2 (n = 2) micrograms/g creatinine, respectively, compared to normal values of 7.5 +/- 0.3 (n = 22) micrograms/g creatinine. Following augmentation therapy, urinary DES values decreased 40 and 36%, respectively, to 11.9 +/- 0.3 (n = 8) and 6.9 +/- 0.4 (n = 7) microgram/g creatinine (p < 0.05). We conclude that monthly AAT augmentation therapy decreased DES excretion in the urine of these PiZ patients. We speculate that since there was lung disease in both patients, a decrease in degradation of lung elastin is the most likely explanation for this observation.

Topics: Adult; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Amino Acids; Bronchiectasis; Desmosine; Elastin; Emphysema; Female; Humans; Isodesmosine; Lung; Male; Middle Aged; Smoking

The current hypothesis of pulmonary emphysema is based on an alteration of the protease-antiprotease balance within the lower respiratory tract. This hypothesis derives largely from studies in emphysema patients with genetic deficiency in serum alpha 1-antitrypsin. In animals, naturally occurring deficiency in serum elastase inhibitory capacity associated with early development of emphysema has been reported in the tight-skin mouse. We describe here a mouse model of genetic deficiency of alpha 1-antitrypsin in which emphysema occurs late in life.. A genetic deficiency in serum alpha 1-antitrypsin was investigated in pallid mice, a strain with spontaneous occurring emphysema. Additionally, the possible pathogenetic role of an elastase-anti-elastase imbalance in pallid mice was investigated using molecular biologic, biochemical, histologic, ultrastructural, and immunoelectron microscopic methods.. Pallid mice have markedly low levels of serum alpha 1-antitrypsin associated with a severe deficiency in serum elastase inhibitory capacity. However, they have normal alpha 1-antitrypsin mRNA levels in the liver. At ultrastructural examination, disruption of alveolar septa is first seen at 8 months of age. At histologic examination, some patchy areas of air-space enlargement with destruction of alveolar septa are seen from 12 months of age onward. These histologic changes are paralleled by a decrease in lung elastin content. The development of the pulmonary lesions is preceded by an alveolar elastolytic burden detected by an immunogold technique.. All these data suggest that the lung changes in pallid mice are the result of an elastolytic process due to a severe inborn deficiency of serum alpha 1-antitrypsin. This animal model reproduces important features of the human condition and may provide new insights into the pathogenesis of emphysema.

Topics: alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Animals; Base Sequence; Disease Models, Animal; Elastin; Lung; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Microscopy, Electron; Microscopy, Immunoelectron; Molecular Sequence Data; Pancreatic Elastase; Pulmonary Alveoli; Pulmonary Emphysema

In recent years a rising interest has developed among the authors studying the problems of pathogenesis and treatment of chronic bronchitis and pulmonary emphysema in the factors responsible for the immune equilibrium of the respiratory system. In particular, the correlation has been stressed between the levels of inflammation mediators and the activity of proteolytic enzymes and their inhibitors in lung tissue. Most studies seem to confirm the validity of the elastase-antielastase concept. Among the environmental factors directly responsible for the development of these diseases tobacco smoke and other irritant fumes (SO2NO2) are mentioned. This concept had been the foundation for trials of therapeutic use of inhibitors of elastolytic enzymes.

Topics: Adult; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Bronchitis; Elastin; Humans; Lung; Middle Aged; Oxygen Inhalation Therapy; Pancreatic Elastase; Pulmonary Emphysema; Smoking

Desmosine is an amino acid specific to elastin. Animal studies suggest that urinary desmosine (UD) represents endogenous elastin degradation. Therefore, UD has previously been used to investigate endogenous elastolysis, but was not elevated in subjects with chronic obstructive airways disease (COAD), although accelerated pulmonary elastolysis is thought to contribute to COAD. We have investigated whether this reflects large day-to-day and between-subject variation in UD and whether, in man, dietary desmosine contributes significantly to that in urine. Mean 24-hour UD output (over 5 consecutive days) from 10 asymptomatic subjects (5 males) was higher in males than females (77.4 +/- 9.6 and 40.2 +/- 5.0 nmol/24 hours, respectively; mean +/- SD, P less than .001), but not significantly different when expressed in terms of creatinine (micrograms desmosine/100 mg creatinine: males, 2.5 +/- 0.4; females, 3.1 +/- 0.8; mean +/- SD). The lowest between-subject variation was observed when the mean of 5 days' 24-hour UD values was analyzed on the basis of gender (coefficient of variation [CV], 12.5%); when gender was not considered, the least between-subject variation was found for the mean of 5 days' desmosine/creatinine analysis (CV, 24.5%). Approximately 1% of dietary desmosine (ingested as [3H]elastin and [3H] desmosine) was excreted in the urine within 24 hours, contributing approximately 15% of UD while on a normal diet. Although ingestion of a low elastin diet (less than 1/10 desmosine/24 hours than a normal diet) resulted in lower within-subject variation in 24-hour UD excretion (mean CV decreased from 31.5% to 20.2%), the between-subject CV and UD levels did not alter.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; alpha 1-Antitrypsin Deficiency; Desmosine; Diet; Elastin; Female; Humans; Injections, Intravenous; Lung Diseases; Male; Middle Aged; Reference Values

Pulmonary elastic fibers in a patient with panacinar emphysema due to alpha-1-antitrypsin deficiency and three patients with centriacinar emphysema related to anthracosis were studied by electron microscopy and by light and electron microscopic immunohistochemistry for elastin. Four types of abnormal elastic fibers were found: (1) finely disrupted fibers, (2) fibers with vacuolar changes and deposits of electron-dense granular material, (3) accumulations of small, rounded amorphous components of elastic fibers near bundles of microfibrils, and (4) large, confluent masses consisting mainly of aggregates of irregularly and compactly arranged, small-sized amorphous components. The amorphous components in these four types of abnormal elastic fibers tended to stain evenly with antielastin antibody. This is attributed to greater penetration of antielastin antibody into fibers that were incompletely polymerized because of immaturity or hydrolytic damage. Finely disrupted fibers were frequently found in the patient with panacinar emphysema and were presumed to have been damaged by elastase. The other three types of elastic fibers were frequently found in the patients with centriacinar emphysema. The vacuoles and electron-dense deposits in elastic fibers probably represented the consequence of damage to elastic fibers. The small round amorphous components in elastic fibers might be formed from abnormal elastogenesis. The large, confluent elastic masses were thought to be formed by the aggregation of elastic fibers in areas of coalescence of alveolar walls undergoing structural remodeling.

Topics: Adult; Aged; alpha 1-Antitrypsin Deficiency; Elastic Tissue; Elastin; Emphysema; Humans; Immunoenzyme Techniques; Lung; Male; Microscopy, Electron; Middle Aged; Models, Biological

Topics: alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Elastin; Humans; Pulmonary Emphysema; Smoking

The effect of repeated intravenous injections of Escherichia coli endotoxin on lung structure and lung parenchymal elastin proportions was studied in rats rendered deficient in alpha 1-antitrypsin by administration of galactosamine. Within 24 h after endotoxin administration, polymorphonuclear leukocyte sequestration was demonstrable by microscopy and differential cell counts of pulmonary lavage fluid. Measurement of the proportions of elastin in lung parenchyma at 24 h revealed values in the normal range; 10 wk after repeated galactosamine and endotoxin administration, there was a reduction in the proportions of lung parenchymal elastin. At 10 wk, these animals showed a significant increase in the mean linear intercept and pulmonary compliance. Animals treated with endotoxin alone developed some but not all of the changes seen in the animals deficient in alpha 1-antitrypsin.

Topics: alpha 1-Antitrypsin Deficiency; Animals; Elastin; Endotoxins; Escherichia coli; Female; Galactosamine; Injections, Intravenous; Lung; Lung Compliance; Pulmonary Emphysema; Rats; Rats, Inbred Strains

Topics: alpha 1-Antitrypsin Deficiency; Elastin; Humans; Lung; Peptide Hydrolases; Pulmonary Emphysema; Smoking

The specific activity of thirteen genetic variants of the protease inhibitor alpha 1-antitrypsin (alpha 1AT) has been determined. Elastase inhibitor activity was assayed protein substrates (elastin and gelatin) and the synthetic substrate N-tert-butoxy-carbonyl-L-alanine p-nitrophenyl ester. The synthetic substrate alpha-N-benzoyl-DL-arginine p-nitroanilide HCl was used to assay trypsin inhibitor activity. The specific activity of alpha 1AT was expressed as serum inhibition/immunological concentration of alpha 1AT. Sera of PI type FM had reduced specific activity with elastase, but not with trypsin. With the possible exception of MP, no other variants showed significant differences in specific activity when compared with normal PI type M.

Topics: alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; alpha-Macroglobulins; Elastin; Gelatin; Genetic Variation; Humans; Pancreatic Elastase; Phenotype; Protease Inhibitors

Insoluble elastin immobilised in Agar plates was used as substrate to quantitate the elastase inhibitory capacity of human sera. This technique was found valuable to evaluate the genetic deficiency of alpha-1-antitrypsin (Z phenotypes) and also the functional loss of the elastase inhibitory capacity in smokers. A significant decrease of elastase inhibitory capacity was demonstrated in the sera of smokers between 40 and 80 years of age, as compared to non smokers of the same age. This method may be useful for the evaluation of patients suffering from elastic tissue diseases (emphysema, arteriosclerosis).

Topics: alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Elastin; Enzymes, Immobilized; Humans; Kinetics; Pancreatic Elastase; Phenotype; Smoking

Topics: alpha 1-Antitrypsin Deficiency; Chemotaxis; Elastin; Humans; Neutrophils; Pancreatic Elastase; Phenotype; Pulmonary Emphysema; Smoking

Human neutrophil elastase degraded tropoelastin approximately 9 times faster than it did solubilized elastin and approximately 19 times faster than it did lung elastin. When bound to alpha2-M, the enzyme retained approximately 6 per cent of its activity toward tropoelastin and solubilized latter observations suggest that alpha2-M--bound elastase, cleared slowly from lung extracellular tissue space, may participate normally in the turnover of soluble precursor (s) of elastin and may contribute to the development of emphysema in alpha1-antitrypsin deficiency.

Topics: alpha 1-Antitrypsin Deficiency; alpha-Macroglobulins; Animals; Dogs; Elastin; Humans; Immunoelectrophoresis, Two-Dimensional; Kinetics; Pancreatic Elastase; Pulmonary Emphysema; Rabbits; Substrate Specificity; Swine; Tropoelastin

The most important primary cause of generalized pulmonary emphysema is in all probability the loss of mechanical stability of the connective tissue framework in the lung parenchyma. The complexity of the interrelations and interdependencies between the fibres and the ground substance, leads to the mapping out of a more detailed outline of the vulnerable parts in a similar framework. It is concluded that the junctions especially constitute weak spots. Glycosaminoglycans, glycoproteins and proteoglycans particularly hold a key position as cementing substances. The importance of female sex hormones in the metabolism of the ground substance is indicated. A survey is given of the possible threats to a similar system. An attempt is made to disentangle the multitude of possible pathogenetic pathways which lead to emphysematous disintegration. Perspectives of future emphysema research are discussed on the basis of these considerations. Preventive, protective, and reconstructive measures are proposed.

Topics: alpha 1-Antitrypsin Deficiency; Ascorbic Acid; Collagen; Connective Tissue; Elastin; Estrogens; Glycoproteins; Glycosaminoglycans; Lung; Lung Compliance; Lysosomes; Microbial Collagenase; Pancreatic Elastase; Peptide Hydrolases; Protein Conformation; Pulmonary Alveoli; Pulmonary Emphysema