Accumulation of eosinophils and T-lymphocytes in the lungs after exposure to pinewood dust.

Exposure to wood dust within the woodworking industry has been shown to cause a variety of respiratory disorders. The aim of this study was to investigate the cellular effects in bronchoalveolar lavage (BAL) fluid and peripheral blood from healthy individuals exposed to pinewood dust. Eleven healthy volunteers were exposed to pinewood dust for 1 h in a whole-body exposure chamber. BAL fluid and blood cells were differentially counted and the expression of activation, adhesion and subset markers on alveolar macrophages and T-lymphocytes was determined 2-6 weeks before and 20 h after the exposure. Following pinewood dust exposure, the total BAL fluid cell concentration increased from 81.4 (64.1-97.5) x 10(6) cells x L(-1) (median (interquartile range)) to 195.3 (154.6-341.2) x 10(6) cells x L(-1). The BAL fluid T-lymphocyte concentration increased from 3.8% (3.5-6.5%) to 7.6% (4.9-11.2%), and BAL fluid eosinophil concentration from 0.0% (0.0-0.2%) to 1.8% (0.6-3.5%). Inhalation of pinewood dust leads to the recruitment of inflammatory cells to the airways of healthy individuals. The increase in numbers of eosinophils, T-lymphocytes and mast cells, i.e. cells of crucial importance to airway inflammation, in the lungs may be related to the increased risk of developing respiratory disorders among woodworkers.

Interstitial lung disease, a common manifestation of newly diagnosed polymyositis and dermatomyositis.

OBJECTIVES: To estimate the prevalence and predictors of interstitial lung disease in newly diagnosed polymyositis and dermatomyositis. METHODS: A prospective study in which consecutive patients with newly diagnosed poly- and dermatomyositis, regardless of clinical symptoms of pulmonary disease, were investigated with chest x ray, high resolution computed tomography (HRCT), pulmonary function tests, and biochemical and autoantibody analysis. Patients with inclusion body myositis, malignancy, other defined inflammatory connective tissue diseases (CTDs), or antibody profile indicating other CTDs were excluded. RESULTS: Between March 1998 and September 2000, 26 new cases of poly- or dermatomyositis were diagnosed; 17 of those patients were included in the study. Interstitial lung disease (ILD), defined as radiological signs on chest x ray examination/HRCT or restrictive ventilatory defect, were found in 11 (65%) patients and were more common in men than in women. Arthritis and occurrence of anti-Jo-1 antibodies were found more often in patients with ILD than in those without. There was no statistically significant association between respiratory symptoms, other serological or laboratory variables and ILD. CONCLUSIONS: ILD is a common early manifestation in patients with poly- and dermatomyositis and is not always related to clinical symptoms. Chest x ray examination, HRCT, pulmonary function tests, and analysis of anti-Jo-1 antibodies should be included in the initial investigation of patients with myositis regardless of respiratory symptoms.

Exosomes with major histocompatibility complex class II and co-stimulatory molecules are present in human BAL fluid.

Exosomes are 30-100 nm diameter vesicles formed by inward budding of endosomal compartments and are produced by several cell types, including T-cells, B-cells and dendritic cells (DC)s. Exosomes from DCs express major histocompatibility complexes (MHC) class I and II, and co-stimulatory molecules on their surface, and can induce antigen-specific activation of T-cells. The aims of the present study were to investigate for the presence of exosomes in bronchoalveolar lavage fluid (BALF) from healthy individuals, and to establish if these exosomes bear MHC and co-stimulatory molecules. The authors analysed BALF taken from seven healthy volunteers and used exosomes from monocyte-derived DC (MDDC) cultures as a reference. After ultracentrifugation, exosomes were bound to anti-MHC class II coated magnetic beads and analysed by flow cytometry and electron microscopy. The authors report for the first time that exosomes are present in BALF. These exosomes are similar to MDDC derived exosomes as they express MHC class I and II, CD54, CD63 and the co-stimulatory molecule CD86. The results demonstrate that exosomes are present in the lung, and since they contain both major histocompatibility complex and co-stimulatory molecules it is likely that they are derived from antigen presenting cells and might have a regulatory role in local immune defence.

Lung accumulations of eosinophil granulocytes after exposure to cornstarch glove powder.

Starch is a main component of wheat flour, which, besides being an occupational allergen can also induce irritative symptoms in the airways. A purified starch product (cornstarch glove powder) was used to investigate whether starch alone could induce airway inflammation. The aim of the study was to investigate a role for starch in wheat flour-induced airway inflammation. Ten healthy individuals were exposed to cornstarch glove powder in a whole-body exposure chamber. Bronchoscopy with bronchoalveolar lavage (BAL) was performed 2-3 weeks before and 1 day after exposure, and the BAL cells were counted differentially. In addition, the expression of activation, adhesion and subset markers on alveolar macrophages and BAL T-cells were investigated using flow cytometry. A three-fold increase in BAL cell concentrations was found, with a selective accumulation and activation of eosinophilic granulocytes, as well as an influx of nonactivated monocytes and polyclonal CD4+ T-cells into the airways. The results show that inhalation of cornstarch glove powder leads to the development of a subclinical inflammation in the airways, with an accumulation of eosinophilic granulocytes. The authors suggest that such exposure may be an interesting model for studying factors contributing to lung accumulations of eosinophil granulocytes in humans.

Effects of formoterol and ipratropium bromide in COPD: a 3-month placebo-controlled study.

The aim of this study was to compare the effects of formoterol, ipratropium bromide and a placebo on walking distance, lung function, symptoms and quality of life (QoL) in chronic obstructive pulmonary disease (COPD) patients. A total of 183 patients (mean age 64 yrs, 86 female) with moderate-to-severe nonreversible COPD participated in this randomised, double-blind, parallel-group study. After a 2-week placebo run-in, patients were randomised to formoterol Turbuhaler 18 microg b.i.d. (delivered dose), ipratropium bromide 80 microg t.i.d. via a pressurised metered dose inhaler, or placebo for 12 weeks. Inhaled short-acting beta2-agonists were allowed as relief medication and inhaled glucocorticosteroids were allowed at a constant dose. The primary variable was walking distance in the shuttle walking test (SWT). Baseline mean SWT distance was 325 m, mean forced expiratory volume in one second (FEV1) was 40% predicted. Clinically significant improvements in SWT (>30 m) were seen in 41, 38 and 30% of formoterol, ipratropium and placebo patients, respectively (not significant). Mean increases from run-in were 19, 17 and 5 m in the formoterol, ipratropium and placebo groups, respectively. Both active treatments significantly improved FEV1, forced vital capacity, peak expiratory flow and daytime dyspnoea score compared with placebo. Formoterol reduced relief medication use compared with placebo. Neither active treatment improved QoL. Formoterol and ipratropium improved airway function and symptoms, without significant improvements in the shuttle walking test.

Rat alveolar and interstitial macrophages in the fibrosing stage following quartz exposure.

Exposure to quartz induces pulmonary inflammation and development of fibrosis. In order to study the fibrosing process, we investigated morphology, function and phenotype of alveolar (AMs) and interstitial (IMs) macrophages at an early stage of fibrosis in rats. Rats were exposed by intratracheal instillations of 10 mg quartz (n=8) or saline (n=8) and studied 3 months later. AMs were obtained by bronchoalveolar lavage and IMs by mechanical fragmentation, followed by enzymatic digestion of lung tissue. Histology revealed subacute silicosis, with early focal fibrosis and alveolar lipoproteinosis. AM quartz exposure increased phagocytic activity and expression of major histocompatibility complex (MHC) Ia antigens, the latter being associated with cellular antigen presenting capacity. IM had an even more pronounced expression of MHC than AM after quartz exposure. Both macrophage fractions had a higher expression of OX-42 (complement receptor 3, CR3) than controls, but the increase in the IM fraction might be explained by the remaining AM in the IM fraction. Exposed AM adhered less to extracellular matrix components (vitronectin and fibronectin) than controls. In contrast, the adhesion of IM to vitronectin increased after exposure. Besides increased adhesion, the effects on IM were scarce. Our results therefore do not support the hypothesis that IM has a key role in the process of inflammation, including fibrosis.

A new whole-body exposure chamber for human skin and lung challenge experiments--the generation of wheat flour aerosols.

A new whole-body exposure chamber for human skin and lung challenge offers possibilities for experimental exposure challenges carried out in clinical practice, for exposure of patients, in research and for investigations of the effects of exposure on the skin and in the respiratory tract. The chamber system can be used for both aerosols and gases. Dynamically controlled, the chamber is relatively easy to operate and to clean. Air exchange rates can be varied between 6-12/h. Initial studies with wheat flour have been carried out. The homogeneity and stability of the wheat flour aerosol concentration (the spatial and the temporal variation) inside the chamber can be kept at acceptable levels.

Inhalation of cold air increases the number of inflammatory cells in the lungs in healthy subjects.

Prolonged exposure to cold air may induce a chronic asthma-like condition in healthy subjects as has been demonstrated in cross-country skiers. In the present controlled study, our aim was to elucidate further the link between cold air exposure and airway inflammation by assessing the cellular influx and mediator levels within the airways following acute exposure to cold air. Bronchoalveolar (BAL) and nasal lavages were performed after exposure to cold air (-23 degrees C) and normal indoor air (+22 degrees C) during a light, intermittent work for 2 h in a cross-over design in eight healthy, nonsmoking, subjects. Analyses of inflammatory cell number, cell activation markers, pro-inflammatory cytokines, albumin and interleukin (IL)-8 in lavage fluids were performed. The number of granulocytes and of alveolar macrophages in BAL fluid was significantly higher after cold air exposure (p<0.05). No increase in BAL fluid lymphocytes and no signs of lymphocyte activation in BAL fluid were found. The concentration of IL-8 was unchanged. There were no signs of granulocyte activation (myeloperoxidase, eosinphilic cationic protein) in BAL fluid. Cold air did not influence the number of inflammatory cells or the concentration of albumin and IL-8 in nasal lavage fluid. In conclusion, exposure to cold air induces an increased number of granulocytes and macrophages in the lower airways in healthy subjects without influencing other inflammatory indices such as cellular activation, plasma leakage and pro-inflammatory cytokines. These findings support the hypothesis that cold air could be of pathogenetic importance in the asthma-like condition previously found in cross-country skiers.

Differences between rat alveolar and interstitial macrophages 5 wk after quartz exposure.

Macrophages play an essential role in pulmonary host defense. We investigated differences between rat alveolar (AM) and interstitial (IM) macrophages after exposure in vivo to quartz, an inducer of intensive alveolitis. Rats were exposed to 0.5 ml of saline without (n = 8) or with (n = 8) 10 mg of quartz by intratracheal instillation. In a third group (n = 8), 10 mg of surfactant were added to the quartz mixture. Five weeks later, AM were recovered by bronchoalveolar lavage and IM by mechanical fragmentation of the lung, followed by enzymatic treatment. Contamination of AM to the IM fraction was calculated to be 12-15%. After quartz exposure, the expression of major histocompatibility complex class Ia was increased in both AM and IM fractions. The receptor corresponding to human complement receptor 3 increased in AM after quartz exposure, and AM from quartz-exposed animals had a lower metabolic activation. Our findings indicate that IM are immunocompetent cells and that differences between AM and IM fractions occur upon quartz-induced inflammation. This response is not affected by addition of surfactant.

Environmental correlates of impaired lung function in non-smokers with severe alpha 1-antitrypsin deficiency (PiZZ).

BACKGROUND: Active smoking is the most important risk factor for pulmonary emphysema in subjects with severe alpha 1-antitrypsin (AAT) deficiency. The aim of this study was to analyse the effects of environmental risk factors other than active smoking on lung function and on respiratory symptoms in non-smoking PiZZ individuals. METHODS: Lifetime exposure to passive smoking, domiciliary use of a kerosene (paraffin) heater or gas cooker, and all occupations since leaving school were reported by 205 non-smoking PiZZ individuals (95 men and 110 women) included in the Swedish AAT deficiency register. Lung function test results and histories of respiratory symptoms (chronic bronchitis, recurrent wheezing, and exertional dyspnoea) were elicited from the AAT register records. RESULTS: After adjustment for age, agricultural employment and domiciliary kerosene heater usage, but not gas cooker usage or passive smoking, were both associated with significantly decreased lung function. Multiple linear regression analysis showed age, sex, kerosene heater usage, and agricultural employment to be independent determinants of lung function impairment. Age and passive smoking for 10 years or more, both at home and at the work place, were associated with the presence of chronic bronchitis. Age and agricultural employment for > or = 10 years were associated with recurrent wheezing and exertional dyspnoea. CONCLUSIONS: Domiciliary kerosene heater usage and an agricultural occupation therefore appear to be environmental factors associated with decreased lung function in non-smoking PiZZ individuals, and passive smoking is associated with an increased frequency of chronic bronchitis, but not with impaired lung function.

Bronchoalveolar lavage findings in firefighters.

Firefighters are exposed to irritating, asphyxiating toxic gases and aerosols. Previous studies indicate that this may result in acute as well as chronic lung function impairment. It is well known that inhalation of organic material can cause alveolitis. The purpose of this study was to investigate whether firefighters repeatedly exposed to fire gases and other combustion products develop inflammatory changes in the lower airways and, if so, what subtype of cells and soluble components accumulate. Thirteen nonsmoking firefighters underwent bronchoalveolar lavage (BAL), and the results were compared to a reference group of 112 nonsmoking healthy volunteers. The firefighters showed lower recovery of instilled lavage fluid (61% vs 72%; P < 0.01) and an increase in the proportion of lymphocytes in that fluid (8.2% vs 5.7%; P < 0.05). Alveolar macrophage fluorescence, which is a sign of uptake of inhaled fluorescent material, rather than a reflection of the degree of inflammation, was increased in two firefighters. Further, the lavage fluid had higher concentrations of native fibronectin (34.6 vs 22.0 micrograms/L; P < 0.01) and hyaluronic acid (27.7 vs 10.0 micrograms/L; P < 0.05). This may indicate local cell activation, since the alveolar capillary membrane probably was intact, as indicated by an unaffected albumin concentration. The observed changes in the deep airways are probably caused by exposure to smoke. It cannot be concluded whether these changes are permanent or are part of a natural temporary defense mechanism.

Functional, morphological, and phenotypical differences between rat alveolar and interstitial macrophages.

Alveolar macrophages (AM) and interstitial macrophages (IM) from rat lungs were characterized with respect to morphology, phagocytosis, adhesion properties, and phenotype. AM were recovered by lung lavage and IM by treatment of the lung tissue with DNAse and collagenase. The AM were enzyme treated in the same way as the IM. The IM were smaller than AM and had a higher nuclear to cytoplasm ratio. They had markedly lower phagocytic capacity. The attachment of particles to the cell surface was significantly lower in IM than in AM, but the capacity to ingest the particles was the same. Adherence to vitronectin- as well as fibronectin-coated surfaces was significantly higher in AM. The phagolysosomal pH was similar in IM and AM, around pH 5, indicating that dissolution of inorganic particles can take place effectively also in IM. Five surface receptors were studied, and the expression differed significantly in all five between AM and IM. The expression of OX-1 (CD 45), a common leukocyte antigen, was significantly higher on AM as was the expression of CD 71 (transferrin receptor). The receptor density for OX-42 was higher on a fraction of IM. This might be compatible with a stronger interaction between these cells and, for example, matrix components. IM had more surface antigen expressing MHC class Ia (OX-6) and CD 54. Both receptors are important for the antigen presentation capacity of macrophages. These findings show profound differences in phenotype between AM and IM and indicate that IM is a highly immunocompetent cell and should not be regarded only as a precursor to AM.

Effect of age and occupational exposure to airway irritants on lung function in non-smoking individuals with alpha 1-antitrypsin deficiency (PiZZ).

BACKGROUND: Severe alpha 1-antitrypsin (AAT) deficiency (PiZZ) is associated with an increased risk of lung emphysema, especially in smokers. The aim of this study was to identify risk factors other than smoking for declining lung function. METHODS: Lung function was studied in 225 self-reported never-smoking PiZZ individuals included in the Swedish AAT deficiency register. RESULTS: Lung function was poorer in men than in women (mean (SD) forced expiratory volume in one second (FEV1) 80 (30) versus 88 (17)% predicted) despite the fact that the men were younger (mean (SD) age 45 (18) versus 51 (17) years), and poorer in those aged 50 or older than in those aged under 50 (mean (SD) FEV1 70 (30) versus 98 (16)% predicted). Self-reported occupational exposure to gas, fumes, or dust occurred more frequently in men than in women. In those aged 50 or older lung function was lower in individuals exposed to airway irritants than those who were not exposed (mean (SD) FEV1 63 (29) versus 76 (31)% predicted). Male sex, increasing age, and previous symptoms of wheezing were independent risk factors for lung function impairment, and male sex, wheeziness, and occupational exposure to airway irritants were independent risk factors in the subjects aged 50 years or more. CONCLUSIONS: In non-smoking PiZZ individuals lung function declines with increasing age, especially after 50. Men are at greater risk of lung function deterioration than women. Asthmatic symptoms and occupational exposure to airway irritants appear to constitute additional risk factors.

Association between the p21 codon 31 A1 (arg) allele and lung cancer.

In previous investigations p53 polymorphisms and haplotypes have been found to be associated with different types of cancer. In this paper the codon 31 polymorphism of the p53-inducible protein p21 was studied in 144 Swedish lung cancer patients and two different control groups: 95 patients with chronic obstructive pulmonary disease (COPD) and 761 healthy controls. An increased frequency of the p21 codon 31 A1 (arg) allele was found in lung cancer patients, especially in comparison with COPD patients (p = 0.004). There was a significantly increased frequency among lung cancer patients of individuals carrying the arg allele both in comparison with COPD controls (OR = 5.2, 95% CI 1.5-18.1) and healthy controls (OR = 1.7, 95% CI = 1.0-2.9). The results of this and previous studies indicate that allelic variants of both p53 and its effector protein p21 may have an influence on lung cancer.

Up-regulation of antigens on alveolar macrophages in quartz exposed rats.

We have analysed the expression of the OX-6 and OX-42 antigens on alveolar macrophages in rats after intratracheal exposure to quartz. Eight female Sprague Dawley rats were included in the study. Four were exposed to physiological saline and four to quartz dust. The rats were sacrificed after 58 +/- 2 weeks and alveolar macrophages were harvested and analysed by flow cytometry. We found a significant higher amount of both antigens on alveolar macrophages harvested from quartz exposed rats. However, we did not find any significant difference in the percentage positive labeled cells between the two groups. We conclude that quartz dust induces up-regulation of functional receptors on alveolar macrophages in rats and that quantitative analysis of these receptors may serve as inflammatory markers in the bronchoalveolar space.

P53 polymorphisms and haplotypes in lung cancer.

An association between the BstU I 1-1 (Pro-Pro) genotype of the p53 codon 72 polymorphism and lung cancer has previously been reported by Kawajiri et al. A reanalysis of the data by Kawajiri et al. revealed no significant difference between patients and controls with respect to allele frequencies, and the increased frequency of BstU I 1-1 homozygotes was mostly ascribable to a deviation from the Hardy-Weinberg equilibrium. In an attempt to replicate the results by Kawajiri et al. we have studied three p53 polymorphisms (BstU I and Msp I RFLPs in exon 4 and intron 6 respectively and a 16 bp duplication in intron 3) and their haplotypes in Swedish lung cancer patients and controls. The results concerning the codon 72 polymorphism were largely negative. Thus there was no significant association between lung cancer and the BstU I 1-1 type, and only a marginal difference (P = 0.044) with respect to the BstU I allele frequency when lung cancer patients were compared with patients with chronic obstructive pulmonary disease (COPD). However, when the analysis was based on haplotype frequencies larger differences appeared and it was found that only BstU I 1 (pro) alleles linked to 16 bp 1 alleles were associated with lung cancer. Pro alleles linked to the 16 bp duplication appeared instead to confer some protection against cancer. Thus the codon 72 alleles need not be functionally involved in lung cancer, but may rather be markers in linkage disequilibrium with other cancer susceptibility sites on p53.

An index of past asbestos exposure as applied to car and bus mechanics.

Different ways to describe historic fibre exposure from asbestos-containing friction materials were studied and compared for a group of 103 car and bus mechanics with more than 20 years employment and 15 years of asbestos exposure. A model was constructed to calculate cumulative asbestos exposure from friction materials including duration, intensity and exposure last year. The model is a combination of an additive and a multiplicative model, where an asbestos index was constructed that takes both near field and far field exposure into consideration. The model was based upon data from the international literature and quantitative asbestos measurements performed 1976-1988 in Swedish car repair workshops. The fibres were counted by phase-contrast microscopy with fibre criteria of length > 5 microns and aspect ratio > or = 3:1. The mechanics' fibre exposure at 398 repair workshops during a period of 48 years were calculated using the model. The mean cumulative exposure was estimated to be 2.6 f ml-1 * year. The annual cumulative exposure was highest for truck mechanics in the early 1960s. The car mechanics had a time-weighted average fibre exposure range of 0.11-0.41 f ml-1 (mean 0.21 f ml-1) in 1965 compared to 0.003-0.08 f ml-1 (mean 0.021 f ml-1) in 1985. In order to validate the model, the mechanic's fibre exposure estimated using the model were compared with representative asbestos exposure measurements for car mechanics during the 1960s and the 1970s (correlation coefficient = 0.69). Five lung physiological variables (FVC, TLC, FEV1, TLco and CV%) were used to study exposure-response relationships. None of the exposure parameters suggested any significant relationship between exposure and decrease in lung function.

Effects within the week on forced vital capacity are correlated with long term changes in pulmonary function: reanalysis of studies on car painters exposed to isocyanate.

OBJECTIVES: To examine if car painters who work with polyurethane paints that contain hexamethylenediisocyanate (HDI) and hexamethylenediisocyanate biuret trimer (HDI-BT) develop acute as well as chronic impairment of lung function. METHODS: In this study data were reanalysed from two earlier studies on a group of car painters to see if a decrease in lung function within the week is a marker of vulnerability in those workers. Data on changes in forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) within the week were available for 20 car painters who were also examined six years later. RESULTS: 10 men showed a decline in FVC within the week. There were no significant differences in age, duration of employment, exposures during the follow up period, or smoking between car painters who had decline in lung function within the week and car painters who had not. A significant correlation was found between the change in FVC within the week and the long term (six year) change in FVC, standardised for the effects of aging and smoking, and adjusted for the number of peak exposures. CONCLUSIONS: The results suggest that the decrease in FVC within the week might serve as a guide to identify car painters at risk of a further decrement in lung function above the effects of aging, smoking, and exposure.

Occurrence of pleural plaques in workers with exposure to mineral wool.

The objective of this study was to investigate whether occurrence of pleural plaques is associated with exposure to mineral wool. The occurrence of pleural plaques on routine chest radiographs of 933 persons employed in the mineral wool manufacturing industry and 865 referents was compared. Twelve men from the mineral wool industry had pleural plaques, as against three of the referents (P > 0.05). The occurrence of pleural plaques among men in the mineral wool industry was not associated with an increased exposure to mineral wool or with length of time between start of employment and time of chest radiograph. The results do not support the notion that inhalation of man-made mineral fibers causes pleural plaques. It must, however, be conceded--keeping in mind the limits imposed by the study size--that nor do they provide grounds for refutation of such a hypothesis.

Sawdust-induced inflammatory changes in rat lung: effects on alveolar and interstitial cells in relation to time.

Exposure to sawdust and its contaminants, e.g., terpenes, may cause respiratory tract and lung parenchymal inflammation. To monitor these changes over time. Sprague-Dawley rats were exposed at one occasion to 2.5 mg sawdust or saline by intratracheal instillation. Flow cytometry analyses were done on bronchoalveolar lavage (BAL) cells. Lung tissue specimens were analyzed histologically and immunohistochemically. After one week, the number of BAL polymorphonuclear leukocytes was increased (P < 0.05, N = 8), followed at six weeks by increases of macrophages and lymphocytes (both P < 0.01, N = 8). Enhanced expressions of class II antigens and complement receptors on macrophages after one week were even more pronounced at six weeks, indicating cellular activation. The BAL findings, also including increased (P < 0.001, N = 8) concentrations of hyaluronan with progressing changes over time, confirmed the signs of inflammation, as did the histological analysis of the lung tissue specimens with an accumulation of polymorphonuclears, macrophages, and hyaluronan in the interstitium.