Human alveolar macrophage phagocytic function is impaired by aggregates of ultrafine carbon particles.

Alveolar macrophages (AM) were collected by bronchoalveolar lavage from healthy volunteers. The AM were loaded with small masses (0.03-3 microg/10(6) AM) of ultrafine carbon particle aggregates. The phagocytic activity of the cells was studied 20 h after the loading. Fluorescein-labeled silica particles (3 microm) were used as test particles and the attachment and ingestion processes were studied separately. In some experiments, AM were incubated with interferon-gamma (IFN-gamma) for 20 h before and during the test of phagocytic activity and during measurement of oxidative metabolism. The ingested carbon particles induced a dose-related impairment of both the attachment and the ingestion processes with a marked impairment down to a carbon particle dose around 0.2 microg/10(6) AM. Such levels should reasonably occur after inhalation of existing concentrations of urban air particles, which to a considerable extent consist of aggregates of ultrafine particles with a carbon skeleton. Incubation with IFN-gamma (12.5 U/ml) also induced significant impairments in both the attachment and the ingestion processes. Loading with carbon further aggravated the effect of IFN-gamma. In contrast to earlier studies in rat AM, IFN-gamma did not impair the oxidative metabolism at rest in these human AM; instead the oxidative metabolism was increased. This difference was due to a difference between rat and human AM and not between rat and human IFN-gamma. Our results suggest that ingested environmental particles in AM, e.g., after an episode of high particle concentration, may impair phagocytic capacity of the cells, especially after infections that induce an increased production of IFN-gamma. Consequently, there might be a risk for additional infections. Moreover, inhaled particles not phagocytized by AM might damage the lung tissue.

Reaction of human alveolar macrophages to exposure to Aspergillus fumigatus and inert particles.

In vitro interaction of human alveolar macrophages (AM) with heat-killed conidia from Aspergillus fumigatus and inert silica particles of similar size, about 3 microns, was studied. The conidia were phagocytized significantly faster by AM than were the control particles partly due to the faster rate of attachment but especially due to the faster rate of ingestion. Quantitative nitroblue tetrazolium (NBT) reduction by AM, reflecting their release of oxygen radicals, was increased by a factor of 2 to 3 in response to the conidia during phagocytosis. The silica particles induced a moderate but significant increase in NBT reduction. Conidia, but not silica particles, showed a considerable percentage (around 8%) of phagolysosomes with neutral pH after 3 h and a smaller percentage (around 1%) after 24 h of incubation. The pH of phagolysosomes with conidia tended to be higher after 3 h, but was significantly lower after 24 h than the pH of phagolysosomes with silica particles. Despite the markedly increased oxidative metabolism there was no increase in cytokine production [interleukins (IL) 6 and 8 and tumor necrosis factor alpha (TNF-alpha)] after exposure to conidia. The silica particles induced a significant decrease in IL-6 and IL-8 production and a tendency toward decreased production of TNF-alpha. The occurrence of phagolysosomes with neutral pH suggests unsealed phagolysosomes from which not only oxygen metabolites but also enzymes might escape from the cell. Lung damage may thus be the result of repeated or long-term exposure to Aspergillus conidia.

Sarcoidosis and regional enteritis in two patients.

Both sarcoidosis and Crohn's disease (regional enteritis) are characterized by granuloma formation and they also have an unknown etiology. Only a few subjects with co-existing sarcoidosis and Crohn's disease have been reported in the literature. We report two cases who suffered from both disorders. In the two subjects Crohn's disease began first and the symptoms of sarcoidosis came later; four and sixteen years, respectively.

The immotile cilia syndrome: characterization of the inflammatory response in nonsmoking patients with dysfunction of the cilia.

In order to demonstrate how patients with immotile cilia syndrome (ICS) are associated with lower respiratory tract inflammation, bronchoscopy and fractionated BAL were performed on eight ICS patients. Their VC was 84.5 +/- 16.7% (mean +/- SD) and FEV1 73.1 +/- 19.9% of predicted. Endobronchial signs of bronchitis were observed in all patients. The total cell concentrations in the BAL fluid were increased, compared to healthy nonsmokers (n = 10), both in the bronchial (BP) and alveolar portion (AP) (p < 0.01 for both). In the BP, this was mainly due to a high concentration of neutrophils (p < 0.001), whereas in the AP, the concentrations of lymphocytes (p < 0.01) as well as all types of granulocytes (p < 0.001-0.01) were elevated. The signs of active inflammation in the lower respiratory tract were confirmed by the concomitantly elevated (p < 0.001-0.05) concentrations of the soluble BAL components albumin, fibronectin and hyaluronan. Thus, the inflammatory response is not restricted to the ciliated conducting airways, but also occurs in the alveolar space and results-surprisingly-in only a slightly impaired lung function.

Phagolysosomal morphology and dissolution of cobalt oxide particles by human and rabbit alveolar macrophages.

The effect of phagolysosomal size on dissolution of cobalt oxide particles was evaluated in two different macrophage systems: alveolar macrophages (AM) of human smokers with phagolysosomes enlarged by ingested cigarette smoke products, and rabbit AM incubated in vitro with sucrose, which causes swelling of the phagolysosomes by osmosis. Human AM from smokers and nonsmokers were studied in vitro. There was no significant difference in particle dissolution between AM obtained from smokers and nonsmokers, although there was a clear difference in the morphological appearance of AM, including significantly larger phagolysosomes in smokers. Rabbit AM were incubated for 24 or 72 h with or without 80 mM sucrose in the medium. The sucrose-treated cells had 3-4 times larger phagolysosomes than untreated cells, with no major change in phagolysosomal pH. The increased size of the phagolysosomes did not affect the ability of the AM to dissolve cobalt oxide particles. Furthermore, rabbit AM showed the same ability as human AM to dissolve the cobalt oxide particles, in spite of the fact that they had markedly smaller phagolysosomes. Another difference between human and rabbit AM was that phagolysosomes in human AMs increased in size with time in culture, while rabbit AM phagolysosomes decreased in size.

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.

Phagolysosomal pH in alveolar macrophages from smokers and non-smokers.

1. Alveolar macrophages efficiently phagocytize particles deposited in the alveolar part of the lung. Phagolysosomal pH in alveolar macrophages from rabbits, dogs, baboons and guinea pigs is around 5 pH units. A low pH is of importance for clearance of inorganic particles and probably also for defence against bacteria and fungi. 2. Phagolysosomal pH in alveolar macrophages from smokers and non-smokers was measured using fluorescein-conjugated amorphous silica particles as a probe. pH was the same in both groups and similar to earlier studies on alveolar macrophages from animals. 3. The morphological picture between alveolar macrophages from smokers and non-smokers differed clearly with a high number of dense inclusions of great variation in size and shape in macrophages from smokers. Ingested test particles were located in all types of inclusions. 4. We conclude that phagolysosomal pH in human alveolar macrophages is similar to that in several animal species and that it is rather insensitive to influence from the environment.

Recurrent Löfgren's syndrome in three patients with sarcoidosis.

Löfgren's syndrome (acute onset of sarcoidosis with erythema nodosum, arthritis, sometimes fever and bilateral hilar lymphoma) is a well known clinical entity. Recurrent Löfgren's syndrome is, however, extremely rare. We report three cases of recurrent Löfgren's syndrome. The duration between the first manifestation of the syndrome and the first relapse varied between two and eight years. One of the patients had two relapses, two and six years after the initial onset of disease.

Terpenes enhance metabolic activity and alter expression of adhesion molecules (Mac-1 and L-selectin) on human granulocytes.

Granulocytes from healthy blood donors were exposed to terpenes dissolved in ethanol. Flow cytometry was used to measure the expression of the cell surface receptors Mac-1 L-selectin, and CR1, which were detected by monoclonal antibodies. The phagocytic activity was determined by using C3b-coated yeast particles as prey. The metabolic activity was measured by determining the intracellular hydrogen peroxide production using dichlorofluorescein diacetate. After terpene exposure the expression of Mac-1 and CR1 increased (P < 0.001 and P < 0.01, respectively) and the expression of L-selectin decreased (P < 0.001). There was also an increased metabolic activity (P < 0.001). The cell viability, the cell count, and the phagocytic activity remained unchanged. These findings suggest that terpene exposure triggers the granulocytes to a higher degree of activation in terms of an altered expression of the adhesion molecules and an increased metabolic activity.

Controlled short-time terpene exposure induces an increase of the macrophages and the mast cells in bronchoalveolar lavage fluid.

Eight healthy nonsmoking volunteers underwent terpene exposure (450 mg/m3 air) on 4 different occasions within a 2 week period. Total exposure time was 12 hours. Before and after the sequences of exposure, bronchoalveolar lavage was performed. Twenty hours after the exposure there was a significant (p < 0.05) increase of the alveolar cell concentration (median pre-exposure value 76 x 10(6) cells/L, post-exposure value 126 x 10(6) cells/L), predominantly due to an increment of the macrophages (from 72 x 10(6) cells/L to 121 x 10(6) cells/L). The mast cells also increased from 1/10 to 5/10 visual fields. No elevated concentrations of albumin, fibronectin, hyaluronan (hyaluronic acid), and tryptase were observed. We conclude that terpene exposure, in a concentration regarded as moderate, induces an acute alveolar cellular reaction in healthy subjects.

Signs of alveolar inflammation in non-smoking Swedish wood trimmers.

Wood trimmers are periodically exposed to mould and may develop extrinsic allergic alveolitis (EAA). To investigate if there were any signs of EAA in wood trimmers with low exposure, 19 non-smoking wood trimmers underwent bronchoalveolar lavage (BAL), spirometry, and measurement of diffusion capacity (TLCO). The group was subdivided into those with (n = 9) and without serological antibodies against mould. In 14 workers the TLCO was measured both at the beginning and at the end of a week at work. Twenty five healthy non-smokers served as BAL controls and 19 healthy non-smokers as lung function controls. The median exposure of total dust was well below the Swedish threshold value, and the exposure of mould and bacteria was also low. The cell concentrations and the proportions of the various alveolar cells did not differ between the groups. The concentrations of the soluble components albumin, fibronectin, and hyaluronan were, however, significantly increased (p less than 0.001 for all) in the workers. No difference was found in lung function between the workers and the controls, and the TLCO was not impaired during a week at work. The groups of seropositive and seronegative workers did not differ in any of these parameters. The results are interpreted as a low intensity alveolar inflammation. The presence of precipitating antibodies against mould did not predict any greater risk of developing a more intense inflammation. Analysis of soluble non-cellular BAL components seems to reflect a discrete ongoing alveolar inflammation better than cell counts only.

Lung function and precipitating antibodies in low exposed wood trimmers in Sweden.

Wood trimmers are exposed to molds that periodically grow on timber that may induce alveolitis and obstructive lung disease. We have evaluated respiratory symptoms, bronchial reactivity, and lung function in 28 wood trimmers at a Swedish sawmill and in 19 unexposed office workers. Eleven (sero-positive) of the wood trimmers had precipitating antibodies in peripheral blood against one or several molds. The exposure to dust (median 0.26 mg/m3), viable mold spores (median 2950 cfu/m3), viable bacteria (median 370 cfu/m3), airborne endotoxins (range 0.0015-0.0025 microgram/m3), and terpenes (range 0.4-23 mg/m3) was lower than levels that earlier have been reported to affect lung function. The wood trimmers reported an increased prevalence of cough and breathlessness. They also showed signs of a mild obstructive impairment with a tendency to increase bronchial sensitivity to metacholine and decreased FEV1 after 2 days free from exposure. FEV1 decreased more during the working week in the sero-positive workers than among the sero-negative workers, and for the whole group the decrease in FEV1 and MEF25 was correlated to the degree of mold exposure.

[Benefits of bronchoscopy in hemoptysis and normal pulmonary X-rays in patients over 40 years of age]. / Bronkoskopi meningsfullt vid hemoptys och "normal lungröntgen" för patienter över 40 år.

Opinion has been divided as to the benefit of bronchoscopy in cases of haemoptysis but normal chest radiograms. In this study such patients were compared with a reference group of haemoptysis patients with X-ray changes. Of the former group 44 underwent bronchoscopy, but after a follow-up of 57 months malignancy has been found in only one case, as compared with a malignancy rate of 47 per cent in the reference group. Thus it is concluded that bronchoscopy is to be recommended in cases of haemoptysis and X-ray changes, whereas non-invasive investigation should suffice in patients under 40 years of age and without X-ray changes, though the risk of malignancy increases with age, particularly in smokers and ex-smokers.