Does cigarette smoke enhance the proteolytic activity of neutrophils?
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Fibronectins↗
Biomedical subjects
K Donaldson
Publications and source records attributed to K Donaldson.
Persistent biological reactivity of quartz in the lung: raised protease burden compared with a non-pathogenic mineral dust and microbial particles.
This study assessed the potential harmfulness of particles in the lung by measuring their ability to elicit and maintain an inflammatory response and to damage lung tissue. It compared the inflammogenicity of two nondurable, biological particulates (Corynebacterium parvum and zymosan) with a pathogenic mineral dust (quartz) and a nonpathogenic dust (titanium dioxide) by dosing rats via the intratracheal route and measuring the consequent alveolitis. The magnitude and duration of the inflammatory response were assessed by measuring the total number of leucocytes and the percentage of neutrophils obtained by bronchoalveolar lavage. Two key functional parameters of the lavaged leucocytes--ability to degrade fibronectin and production of plasminogen activator--were also measured. A marked inflammatory response had occurred by one day after instillation, characterised by increases in total leucocyte numbers and percentage of neutrophils in the bronchoalveolar lavages, with all four test materials. In all but the quartz exposed animals, the inflammation subsided rapidly thereafter, approaching control levels by 15 days after injection; in the quartz exposed animals the alveolitis persisted for up to 30 days. All of the inflammogens generated chemotaxins in rat serum in vitro and so, by analogy, might also be expected to generate chemotactic activity in alveolar lining fluid which could contribute to the generation of an inflammatory response. The cellular inflammatory response was accompanied by a concomitant increase in the proteolytic activity of the bronchoalveolar lavage leucocytes but production of plasminogen activator remained unchanged. In vitro exposure to the inflammogens had no effect on the proteolytic activity against fibronectin or on the plasminogen activator activity of bronchoalveolar leucocytes.
Animals↗
Injurious effects of wool and grain dusts on alveolar epithelial cells and macrophages in vitro.
Epidemiological studies of workers in wool textile mills have shown a direct relation between the concentration of wool dust in the air and respiratory symptoms. Injurious effects of wool dust on the bronchial epithelium could be important in causing inflammation and irritation. A pulmonary epithelial cell line in vitro was therefore used to study the toxic effects of wool dust. Cells of the A549 epithelial cell line were labelled with 51Cr and treated with whole wool dusts and extracts of wool, after which injury was assessed. Also, the effects of grain dust, which also causes a form of airway obstruction, were studied. The epithelial injury was assessed by measuring 51Cr release from cells as an indication of lysis, and by monitoring cells which had detached from the substratum. No significant injury to A549 cells was caused by culture with any of the dusts collected from the air but surface "ledge" dust caused significant lysis at some doses. Quartz, used as a toxic control dust, caused significant lysis at the highest concentration of 100 micrograms/well. To determine whether any injurious material was soluble the dusts were incubated in saline and extracts collected. No extracts caused significant injury to epithelial cells. A similar lack of toxicity was found when 51Cr labelled control alveolar macrophages were targets for injury. Significant release of radiolabel was evident when macrophages were exposed to quartz at concentrations of 10 and 20 micrograms/well, there being no significant injury with either wool or grain dusts. These data suggest that neither wool nor grain dust produce direct injury to epithelial cells, and further studies are necessary to explain inflammation leading to respiratory symptoms in wool and grain workers.
Animals↗
Leukocyte-mediated epithelial injury in ozone-exposed rat lung.
Both epithelial injury and inflammation are characteristic findings in the centriacinar regions of the lungs of rats exposed to ozone. In humans such effects could lead to long-term lung damage and disease. In animals, neoplastic change in the lungs after exposure to ozone has been described previously. The possible relationships between inflammatory cell recruitment, epithelial injury, and hyperplasia, with special regard to the important role of repair processes in leading to increased incidence of tumors in some species, have been addressed in the present study. We have previously described that leukocytes from lungs inflamed by different agents can injure epithelial cells in vitro. We have suggested that this leukocyte-mediated epithelial injury could enhance epithelial turnover in ozone-exposed lungs and so enhance the likelihood of tumor development. We, therefore, set out to test the hypothesis that bronchoalveolar leukocytes from ozone-exposed lungs can injure epithelial cells in vitro. PVG rats were exposed to 0.2, 0.4, 0.6, and 0.8 parts per million2 (ppm) ozone for seven hours per day for up to four days. On the morning following the last exposure, bronchoalveolar lavage was used to sample the bronchoalveolar leukocytes and the following parameters were assessed: total number, differential leukocyte count, production of oxidants, ability to degrade fibronectin, and ability to injure epithelial cells. In addition to these parameters, which were measured at all concentrations and time points in limited experiments, we also assessed macrophage size in short-term culture and inflammation in histological sections of lungs. Total number of lavageable cells was not affected by ozone inhalation. However, the percentage of macrophages decreased with ozone treatment and the percentage of neutrophils increased on days 1 and 2 at 0.6 and 0.8 ppm ozone. There was no significant effect of ozone exposure on the ability of neutrophils to degrade fibronectin or injure epithelial cells. Production of superoxide anion in response to stimulation with phorbol myristate acetate was significantly decreased by exposure to 0.6 ppm ozone, as described in previous studies. Macrophages from the lungs of rats exposed to ozone were larger than control macrophages.
Animals↗
Lack of effect of N-acetylcysteine on the release of oxygen radicals from neutrophils and alveolar macrophages.
N-acetylcysteine (NAC) is rapidly de-acetylated in vivo to cysteine (CYSH), a precursor of glutathione (GSH) which is an antioxidant in cells and body fluids. We investigated the effect of oral administration of N-acetyl cysteine for 5 days on the spontaneous and stimulated generation of hydrogen peroxide (H2O2) and superoxide anion (O2-) from human and rat phagocytic leucocytes. Alveolar macrophages (AM) were obtained by bronchoalveolar lavage (BAL) in control rats and rats given NAC in their drinking water. Neutrophils (PMNL) were harvested from whole blood in normal nonsmoking volunteers before and after NAC was given by mouth. The stimulated release of H2O2 and O2 from both rat AM and human PMN was not changed by administration of NAC. However, a small but significant increase was observed in both the spontaneous generation of O2- from rat AM and the spontaneous generation of H2O2 from human PMNL. Administration of NAC significantly increased cysteine levels in human plasma and rat BAL, but the levels in human PMNL and rat AM after NAC did not differ from control levels. GSH levels were not altered significantly by NAC.
Acetylcysteine↗
Raynaud's syndrome.
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Angiotensin-Converting Enzyme Inhibitors↗
Impaired chemotactic responses of bronchoalveolar leukocytes in experimental pneumoconiosis.
Rats were exposed to clouds of the following pneumoconiotic dusts: quartz, coal-mine dust, and chrysotile asbestos at 10 or 50 mg/m3 for 8, 32, and 75 days; for comparison, rats were also exposed to the non-pathogenic dust titanium dioxide (TiO2). The bronchoalveolar leukocytes (macrophages and neutrophils) from dust-exposed and control rats were obtained by lavage and tested for their ability to migrate toward zymosan-activated serum. Varying amounts of neutrophils were present depending on the ability of the dust to cause inflammation and the length of exposure. There was a marked loss of chemotactic ability in leukocytes from rats inhaling the pneumoconiotic dusts compared with controls; TiO2-exposed leukocytes had some impairment of chemotaxis, but this was substantially less than that found with the pneumoconiotic dusts. The loss of chemotactic activity did not correlate with the percentage of neutrophils in the lavage cells except when there were very high levels of neutrophils, and there was substantial impairment of chemotaxis with negligible numbers of neutrophils, showing that macrophage chemotaxis was impaired. A phagocytic burden within the leucocytes was not sufficient alone to inhibit chemotaxis, nor was the loss of chemotactic activity due to occupied receptors, since incubation failed to restore chemotaxis. Loss to chemotactic activity by leukocytes from pneumoconiotic dust-exposed lung could be an important factor in the development of pneumoconiosis.
Animals↗
Secretion of plasminogen activator inhibitor by normal rat pleural leukocytes in culture.
The normal balance between coagulation and fibrinolysis in the pleural cavity is poorly understood despite the critical role of the pleura in the movement of the lungs. To determine the fibrinolytic activity and the interaction between plasminogen activators and their inhibitors in the normal pleural space, we tested normal rat pleural leukocytes, principally macrophages and mast cells, and their supernatants, for activity in an [125I]fibrin degradation assay. It was found that pleural leukocytes did not release plasminogen activator, but the leukocytes and their supernatants inhibited the plasminogen-dependent fibrinolysis caused by both alveolar leukocytes and mesothelial cells. Further experiments demonstrated that pleural leukocytes produce a protein inhibitor primarily against urokinase-induced fibrinolysis in culture and that macrophages are the main source of the inhibitor. The lysate of mast cell-enriched population exhibited high plasminogen activator activity while no such activity could be determined in macrophage-enriched lysate. These data show that normal rat pleural leukocytes contain plasminogen activator inside the cells and synthesize a urokinase-type plasminogen activator inhibitor in culture that may be important in the fibrinolysis/coagulation balance in the pleural space.
Animals↗
Alveolitis caused by exposure to coal mine dusts: production of interleukin-1 and immunomodulation by bronchoalveolar leukocytes.
Two kinds of coal mine dust, low rank with high quartz (bituminous) and high rank with low quartz (anthracite), were assayed for ability to induce alveolitis and to stimulate interleukin-1 release from normal alveolar macrophages in vitro. Dust-elicited bronchoalveolar leukocytes were also assessed for their effects on macrophage-depleted splenocyte mitogenesis and their ability to produce interleukin-1. Quartz and titanium dioxide were used for comparison as toxic and inert dusts, respectively. Both coal mine dusts caused substantial release of interleukin-1 from normal alveolar macrophages in vitro and the levels were higher than those caused by quartz. Bituminous coal mine dust provoked acute but rapidly subsiding macrophage/neutrophil alveolitis which was greater than that provoked by titanium dioxide; anthracite caused less alveolitis than titanium dioxide and quartz caused large scale, persistent alveolitis. Whole bronchoalveolar leukocytes, including polymorphonuclear neutrophils, elicited by exposure to dust, were less inhibitory to lymphocyte mitogenesis than normal alveolar macrophages and bituminous coal mine dust-induced neutrophils were augmentary to lymphocyte mitogenesis; macrophages from inflamed lungs were, on the whole, inhibitory to lymphocyte mitogenesis. Alveolar macrophages from bituminous coal mine dust- or titanium dioxide-exposed lungs showed increased ability to release interleukin-1 on stimulation in vitro. These findings suggest that bronchoalveolar leukocytes elicited by coal mine dust could modulate immunity by means of interleukin-1 release and enhancement of lymphocyte proliferation.
Animals↗
Contrasting bronchoalveolar leukocyte responses in rats inhaling coal mine dust, quartz, or titanium dioxide: effects of coal rank, airborne mass concentration, and cessation of exposure.
The aim of this study was to determine the bronchoalveolar leukocyte response to airborne coal mine dust; quartz and titanium dioxide were used as positive and negative controls, respectively. Groups of rats were exposed to airborne mass concentrations of 10 and 50 mg/m3 of the dusts for 7 hr/day, 5 days/week and their bronchoalveolar space was lavaged at time points between 2 and 75 days of exposure, to assess the leukocyte response. This study revealed time-dependent and airborne mass concentration-dependent recruitment of neutrophils and macrophages into the bronchoalveolar region with coal mine dust inhalation but no real difference in the magnitude of the response between coal mine dusts from collieries mining coal of different rank and quartz content although the maximum quartz content in the dusts used was 6%. The inflammatory response was much less than that produced by quartz, at similar airborne mass concentrations, and more than that produced by titanium dioxide which was, in general, a poor inflammogen in the rat lung. Groups of rats were exposed to the airborne dusts for 32 or 75 days, then removed from the exposure chambers, and allowed to recover by breathing room air for a further 64 days. During this recovery period there was marked progression of the leukocyte response with quartz and persistence of the response with coal mine dust. Chronic recruitment of leukocytes to the lungs of individuals inhaling coal mine dust is likely to be an important factor in the development of coal workers' pneumoconiosis.
Administration, Inhalation↗
Immunomodulation in mineral dust-exposed lungs: stimulatory effect and interleukin-1 release by neutrophils from quartz-elicited alveolitis.
Quartz deposition in the rat lung causes an intense and persistent neutrophil alveolitis leading to parenchymal fibrosis. Bronchoalveolar leucocytes (BAL) from quartz-exposed rat lungs were studied for their effects on splenic lymphocyte proliferation; titanium dioxide (TiO2) was used as a control, non-fibrogenic dust. Seven days after the intratracheal instillation of 1 mg of quartz or TiO2 suspended in phosphate-buffered saline (PBS), BAL were recovered by lavage; the effect of PBS alone was also studied. TiO2-elicited BAL (macrophages greater than 98%) inhibited splenocytes responding to suboptimal phytohaemagglutinin (PHA) more than PBS-elicited BAL (macrophages greater than 98%); the effect was dependent on the BAL:splenic lymphocyte ratio. Quartz-elicited whole BAL (macrophages 49%, neutrophils 51%), and an alveolar macrophage-enriched population with purity of 87% separated from it, were less inhibitory to splenocyte mitogenesis than PBS-elicited BAL. A neutrophil-enriched population, with a purity of 80%, markedly enhanced splenocyte response to PHA. In addition, whole quartz BAL and the macrophage-enriched population obtained from it enhanced the mitogenesis of T cell-enriched lymphocytes at a much lower BAL:lymphocyte ratio. The neutrophil-enriched quartz BAL enhanced mitogenesis substantially more than the whole or macrophage-enriched population from quartz-exposed lung. Supernatants from normal macrophages, PBS BAL, TiO2 BAL, quartz BAL and both alveolar macrophage and neutrophil-enriched quartz populations were assessed for interleukin-1 (IL-1) activity. Quartz-BAL, quartz macrophages and quartz neutrophils all produced significantly higher IL-1 levels than PBS BAL; the supernatants from quartz neutrophils, however, showed the highest IL-1 activity. These findings suggest that quartz-elicited bronchoalveolar leukocytes, especially neutrophils, enhance lymphocyte proliferation and that increased IL-1 secretion by these cells is likely to be the effector molecule involved. These findings have important implications for immune response in mineral dust-stimulated lung and for inflammatory lung disease in general.
Animals↗
Lymphocyte modulation by inflammatory bronchoalveolar leukocytes.
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Animals↗
Inflammation in the lungs of rats after deposition of dust collected from the air of wool mills: the role of epithelial injury and complement activation.
In a previous study assessing respiratory symptoms in individuals employed in wool textile mills in the north of England relations between symptoms of chronic bronchitis, breathlessness and wheeze, and rhinitis and current exposure to airborne mass concentration of dust were shown. As preliminary steps in defining the potential hazard associated with dust from the air of wool mills the ability of inspirable dust, collected from the air of wool textile mills, to cause inflammation when injected into the lungs of rats was determined. Dusts were collected from the beginning of wool processing (opening) in one factory and from the middle (combing) and late (backwinding) stages of the process in two other factories. Ability of the dusts to cause inflammation was assessed by instillation into the lungs of rats followed by bronchoalveolar lavage. All the dusts caused some inflammation which peaked on day 1 and did not persist beyond one week. A distinctive aggregation response of mononuclear cells in the lavage, however, had a different time course, peaking at day 7. An attempt was made to determine how the wool mill dusts caused inflammation and experiments showed that the dusts themselves had no inherent chemotactic activity but that they did have a pronounced ability to generate chemotaxins in serum and so could activate complement in lung fluid. In addition, dust collected from ledges in the mills had the ability to injure epithelial cells in vitro which could also contribute to inflammation. A role for endotoxin in the inflammatory activity of the dusts was not discounted and a leachate of the dust had the ability to cause inflammation when injected into the lungs of rats. Wool mill dust is likely to be a complex mixture of materials and these experiments represent a preliminary approach to understanding the biological activity of the whole unfractionated dust and further studies are in progress to define more accurately the toxic material(s) in the dust.
Animals↗
A dosimetric approach for relating the biological response of the lung to the accumulation of inhaled mineral dust.
Results from studies of the retention of contrasting mineral dusts inhaled by rats (for periods of up to three months) and the resultant changes in the phagocyte defence system of the deep lung were examined. The dusts used were titanium dioxide (relatively innocuous) and quartz (relatively toxic). The parameters assessed included the accumulation of material in the lung and lymph nodes during chronic exposure and the associated leukocyte response as assessed by broncho-alveolar lavage. The principal findings were that: (a) low level exposure to titanium dioxide produced no measurable inflammation (as indicated by neutrophil recruitment) but higher concentrations (30, 50, and 90 mg/m3) caused the transfer of dust to lymph nodes and first evidence of inflammation; and (b) for quartz, there was a more prominent response and earlier transfer of material to the lymph nodes. The suggested relation between changes in the neutrophil population and dust accumulation is discussed in terms of a quantitative dosimetric model, from which implications for assessing and managing human exposures emerge.
Animals↗
Production of interleukin-1 like activity by neutrophils derived from rat lung.
Interleukin-1 like activity was produced by neutrophils obtained by bronchoalveolar lavage from experimentally inflamed rat lung. Activity was released spontaneously from neutrophils at high levels but it was enhanced by stimulation with endotoxin in vitro.
Animals↗
Effects of products from inflammatory pulmonary neutrophils on alveolar macrophage chemotaxis, spreading, and thymidine incorporation.
Since macrophages and neutrophils are found together in the alveolar region of the lung during alveolar inflammation, we assessed whether neutrophil products could influence three key functions of alveolar macrophages: chemotaxis, spreading, and thymidine incorporation. Neutrophils were obtained from the lungs of rats treated by intratracheal instillation of heat-killed Corynebacterium parvum and cultured overnight, alone or in the presence of zymosan, PMA, an inert particulate (titanium dioxide), or a toxic dust, (quartz). Supernatants were collected from these cells and a lysate, obtained by freeze-thawing neutrophils, was also used. Neutrophil supernatants caused a slight reduction in chemotaxis and a significant loss of ability to spread on glass which varied depending on the in vitro treatment of the neutrophils. In addition neutrophil supernatants also had a substantial effect in stimulating uptake of thymidine which was, once again, very dependent on the treatment of the neutrophils during preparation of the supernatants, with unstimulated and TiO2-treated neutrophils producing maximum stimulation. The increases in thymidine uptake were not matched by increased proliferation, suggesting that another signal may be necessary for expanison of alveolar macrophage numbers during alveolar inflammation.
Animals↗
Bronchoalveolar leukocyte response in experimental silicosis: modulation by a soluble aluminum compound.
The biological properties of quartz have been related to its surface reactivity. We have addressed the role of particle surface reactivity in mediating the biological activity of quartz in mixed dusts, by treating the quartz with aluminum lactate. Intratracheal instillation of untreated quartz in rats caused a rapid, sustained alveolitis and bronchoalveolar leukocytes obtained from these animals had enhanced activity in degrading fibronectin, but reduced ability to mount a respiratory burst. Quartz pretreated with aluminum elicited a markedly reduced inflammatory response; the reduced activity of the treated quartz was also reflected in the attenuated change in the key functional parameters, oxidant production and proteolysis of fibronectin. Late intratracheal dosing with aluminum after the quartz-induced alveolitis was well established reduced the inflammatory response and abrogated the effect of quartz on the respiratory burst, but did not alter fibronectin degradation by the leukocytes. Aluminum did not affect the inflammatory response to Corynebacterium parvum and thus the effect was on the quartz particles and not on the inflammatory leukocytes. These findings have implications for the likely pulmonary responses to mixed dusts containing quartz and aluminum silicate clays.
Animals↗
Phagocyte-derived oxidant and protease in mineral dust-related lung disease.
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Animals↗