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Biomedical subjects

L Belin

Publications and source records attributed to L Belin.

At least 19 recordsLinked to original sources

Hyaluronic acid (hyaluronan) in BAL fluid distinguishes farmers with allergic alveolitis from farmers with asymptomatic alveolitis.

Pulmonary function measurements, bronchoalveolar lavage (BAL), and analyses of precipitating antibodies in blood were performed in 12 farmers wtih no symptoms from the airways and 12 farmers who were admitted to the hospital due to acute symptoms of alveolitis (all nonsmokers). In addition, a bronchial methacholine provocation test was performed in the asymptomatic farmers. In 11 of the 12 symptomatic farmers but in none of the asymptomatic farmers, precipitating antibodies against one or more of the microorganisms which usually occur in a farmer's environment were found. In the farmers with symptomatic alveolitis, a restrictive impairment of pulmonary function was found, while pulmonary function was normal in all asymptomatic farmers. Findings in the BAL fluid showed increased concentrations of total cells, lymphocytes, and neutrophils and elevated levels of albumin, fibronectin, and angiotensin-converting enzyme in asymptomatic farmers compared with our own reference group. The same analyses in BAL fluid from the symptomatic farmers revealed a further increase in all parameters compared with the asymptomatic farmers. The BAL fluid from asymptomatic farmers had normal levels of hyaluronic acid (hyaluronan) and procollagen 3 N-terminal peptide, while these levels were significantly increased in the symptomatic group. We conclude that inflammation in the alveolar space and signs of activation of alveolar macrophages are present in farmers regardless of respiratory symptoms, although these findings are more pronounced in the presence of symptoms of acute alveolitis; however, the findings of impaired pulmonary function and the occurrence of precipitins and elevated levels of hyaluronic acid and procollagen 3 N-terminal peptide in BAL fluid were exclusively found in the farmers with airways symptoms. We postulate the hyaluronic acid, due to its pronounced ability to immobilize water, may be of importance in the development of the pulmonary function impairment observed in farmer's lung disease.

Acute Disease

Alterations in bronchoalveolar lavage fluid but not in lung function and bronchial responsiveness in swine confinement workers.

Testing of lung function and bronchial reactivity, bronchoalveolar lavage (BAL), and a skin prick test with a standard panel and six "swine" extracts obtained from swine and swine environment were performed in 20 randomly selected nonsmoking swine confinement workers. In addition, blood samples for detection of antibodies by the diffusion in gel-enzyme-linked immunosorbent assay (DIG-ELISA) technique and precipitating antibodies were drawn. Air samples for measurement of dust and endotoxin levels were collected. All the farmers regarded themselves as healthy. The results were compared with reference groups consisting of urban nonsmoking subjects who had not been exposed to pig farming environment. The pig farmers had normal lung function and the bronchial reactivity was not different from the reference group. In the BAL fluid of the farmers, the concentration of total cells and granulocytes was increased while the concentrations of lymphocytes and macrophages were normal. The BAL fluid concentrations of albumin, fibronectin, and hyaluronan were elevated in the farmers. Skin prick tests with swine extracts were negative in all farmers. Antibodies (assessed by DIG-ELISA) against swine dander, swine dust, and pig feed were increased and precipitating antibodies against swine dander were found in 14, against pig food in five, and against swine confinement dust in three of the 20 pig farmers. The concentration of airborne total dust was 7.4 mg/cu mm and the endotoxin concentration was 37 (22 to 60) ng/cu mm during tending the pigs and increased, during feeding, to 13.8 mg/cu mm and 315 (194 to 716) ng/cu mm, respectively. There was no correlation between exposure and lung function or lavage findings. In conclusion, randomly selected pig farmers had signs of airway inflammatory reaction and activation of the immune system without alteration in lung function and bronchial reactivity.

Adult

Clinical and immunological responses to occupational exposure to alpha-amylase in the baking industry.

alpha-Amylase is a starch cleaving enzyme often used in the baking industry as a flour additive. It is usually of fungal origin, produced by Aspergillus oryzae. One previous report has shown IgE antibodies and positive skin prick test against alpha-amylase in asthmatic bakers. This paper describes four alpha-amylase sensitised index cases with occupational asthma or rhinitis and the results of a cross sectional study of 20 workers from the same factory who were also exposed to alpha-amylase powder. Air sampling detected airborne alpha-amylase at a concentration of 0.03 mg/m3. Significantly more work related symptoms such as rhinitis and dermatitis were found among the alpha-amylase exposed workers compared with referents. A skin prick test to alpha-amylase was positive in 30% (6/20) of the exposed workers. Most of the persons showing a positive skin prick test had work related symptoms and were also skin prick test positive to common allergens. Nasal challenge tests with amylase were performed in selected cases and validated three cases of alpha-amylase induced rhinitis. Two non-symptomatic workers had precipitins to alpha-amylase. Specific IgG antibodies were shown by two further serological techniques. The nature and relevance of these antibodies are currently being studied. It is concluded that alpha-amylase powder is a potent occupational sensitiser. Precautions should be taken when handling this allergenic enzyme.

Adult

An optimized assay of specific IgE antibodies to reactive dyes and studies of immunologic responses in exposed workers.

Methods of assaying reactive dye-specific IgE antibodies were investigated with a RAST. Sera from three patients, occupationally exposed to a reactive dye, Remazol black B (Chemical Abstract registry number 17095-24-8), were used. Directly dyed disks, that is, disks without any carrier protein, resulted in poor and unreliable measures of specific IgE. In contrast, optimized preparation of conjugates between the dye and human serum albumin resulted in efficient binding of specific IgE. The patients' RAST results were strongly positive, whereas sera from 36 exposed workers but without symptoms and sera from unexposed subjects with high levels of total IgE were negative. The hapten and carrier specificity of the IgE antibodies was studied by direct RAST and RAST inhibition. In one patient, the antibodies were principally hapten specific, whereas another patient was found to have antibodies with a high degree of specificity to the carrier. The third patient's antibodies were intermediate between the other two patients' antibodies in this respect, suggesting that antibody specificity is dependent not only on the nature of the hapten but also on individual immune response factors. The study demonstrates that it is important to use an optimized preparation of dye-protein conjugates to elicit reliable results and a high degree of specific IgE binding in the RAST.

Antibody Specificity

An in vitro method for predicting sensitizing properties of inhaled chemicals.

A method for monitoring chemical reactivity in aqueous solutions, at neutral pH and 37 degrees C, was developed. The chemical was allowed to react with a lysine-containing peptide, and the reaction was monitored with high-performance liquid chromatography. Simple acids, bases, and solvents did not react with the peptide, whereas isocyanates, anhydrides, and chloramine-T, substances well known for their sensitizing and asthma inducing properties, did. Thus a positive test strongly suggested that the chemical had the potential to act as a hapten and cause sensitization when inhaled. Prepolymers of diphenylmethane diisocyanate were considerably more reactive than prepolymers of toluene diisocyanate or hexamethylene diisocyanate. Isocyanates blocked with caprolactam, butanone oxime, malonic acid diethylester, or isononyl phenol showed no reactivity. This result suggested a significantly reduced risk of respiratory reactions when such blocked isocyanates are handled at room temperature. One blocked isocyanate showed, however, considerable reactivity.

Administration, Inhalation

Immunologic specificity of isocyanate-induced IgE antibodies in serum from 10 sensitized workers.

A procedure for the preparation of RAST disks used to assay isocyanate-specific IgE antibodies was developed. The outcome of the RAST was found to be strongly dependent on how the isocyanate test antigen was synthesized. Specific IgE from selected workers with isocyanate asthma reacted optimally to conjugates with less than or equal to 10 isocyanate molecules bound per molecule of human serum albumin. Further haptenization resulted in decreased specific binding and increased nonspecific binding because of high levels of total IgE. The hapten and carrier specificity of isocyanate-induced IgE antibodies were studied by direct RAST and RAST inhibition. The existence of new antigenic determinants related to both the isocyanate hapten and the carrier could be demonstrated. It was important to use a test antigen prepared from the same isocyanate as that to which the worker had been exposed, since the cross-reactivity between different isocyanate haptens was partial and varied from one patient to another. It was confirmed that isocyanate-specific IgE antibodies can be demonstrated only in a subgroup of workers with isocyanate-related bronchial asthma.

Antibody Specificity

Immunological specificity of chloramine-T-induced IgE antibodies in serum from a sensitized worker.

A procedure for the preparation of chloramine-T (CT) conjugates used to assay IgE antibodies was developed using response surface methodology and serum from a subject occupationally exposed to the substance. The conjugates, synthesized by reacting CT with human serum albumin (HSA) and other protein carriers, were used as antigens in a radio-allergosorbent test (RAST). Human serum albumin was found to be a suitable carrier, although other protein carriers also gave specific IgE-binding of a similar extent. The CT-HSA conjugates used in the RAST were characterized by high performance liquid chromatography, electrophoresis, immunodiffusion and ammonium sulphate precipitation. However, no strong correlation was seen between the ability of the conjugates to bind IgE and their physical or immuno-chemical properties. The hapten and carrier specificity of CT-induced IgE antibodies in the subject's serum were studied by direct RAST and RAST inhibition. No existence of new antigenic determinants related to the carrier could be demonstrated. Although HSA as a carrier was altered immunochemically by CT, the IgE antibodies were found to be specific to hapten only. Chloramine-T-specific IgG antibodies could not be demonstrated in the subject's serum.

Adult

Environmental and health studies of farm workers in Swedish swine confinement buildings.

The relation between the health of workers and the environment in swine confinement buildings was investigated in a study of 57 workers on 30 swine farms in southern Sweden and 55 matched controls. Swine workers reported significantly higher frequencies of respiratory symptoms, more frequent colds and absence due to chest illness, and a history of pneumonia. The increased frequency of symptoms of respiratory disease was related to the number of years and percent of the day spent working with swine. Symptoms were also associated with respirable dust, total dust, endotoxin in total dust, and number of microbes in the air of the work environment. In a multiple regression analysis of the relation between 16 different environmental parameters to work period shifts of five pulmonary function parameters, endotoxin was found to be significantly related to the FEV1 in a dose dependent way.

Agricultural Workers' Diseases

Isocyanate exposure and hypersensitivity pneumonitis--report of a probable case and prevalence of specific immunoglobulin G antibodies among exposed individuals.

A car painter experienced three episodes of a hypersensitivity pneumonitis-like disease after exposure to two-component acrylic lacquers with hexamethylene diisocyanate (HDI) as the curing agent. High titers of HDI-specific immunoglobulin (Ig) G antibodies were found in the patient's serum by means of enzyme-linked immunosorbent assay (ELISA). In the ELISA, 5 to 10% of the sera from 455 isocyanate-exposed but asymptomatic workers were positive, depending on the criterion used for a positive test, whereas 0% of the sera from 157 unexposed referents was found to be positive. Among 10 subjects with isocyanate-induced asthma and isocyanate-specific IgE antibodies, 50% had specific IgG. It was concluded that the presence of isocyanate-specific IgG antibodies in serum is correlated with isocyanate exposure rather than with symptoms of isocyanate-induced disease.

Adult

Longitudinal study of specific IgE and IgG antibodies in a patient sensitized to ethylene oxide through dialysis.

IgE and IgG antibodies to ethylene oxide (EO) were monitored by RAST and by ELISA in serum from a patient with hypersensitivity reactions during hemodialysis. Serum samples from the patient covering a 7-year period were analyzed. The changes in titers of IgE and IgG antibodies correlated to the time of EO exposure as well as to clinical symptoms. EO-specific IgG antibodies were, however, also found in sera from hemodialysis patients without any hypersensitivity symptoms. Assay of specific IgG antibodies will only indicate whether a patient has been exposed to, and immunostimulated by, EO, whereas specific IgE antibodies appear to be clearly related to hypersensitivity symptoms. The hapten and carrier specificity of EO-induced IgE antibodies was studied. The antibodies were found to be solely hapten specific because carriers of different types could be used in RAST without changing the outcome of the test. The existence of new antigenic determinants related to the carrier could not be demonstrated.

Adult

Breed-specific dog-dandruff allergens.

Fifty-one patients with clinical history of dog allergy were skin prick tested with eight individual standardized dog breed-allergen preparations, one mixed breed-allergen preparation (Poodle/Alsatian), dog-serum albumin, and histamine hydrochloride, 1 mg/ml. All extracts were characterized by crossed immunoelectrophoresis and crossed radioimmunoelectrophoresis with a pool of sera from patients clinically sensitive to dog. The dog-breed extracts contained common antigens/allergens, as well as components represented only in one or two dog-breed extracts. The concentration corresponding 1000 BU/ml varied from 16 to 100 micrograms of protein per milliliter. The sensitivity of skin prick test was 67% to 88% for the various dog breed-allergen preparations, but only 18% for dog-serum albumin. Significant difference between the skin test response to different dog breed-allergen preparations indicating dog breed-specific allergens was obtained in 15% of the patients. There was no significant correlation between skin prick test results and symptoms related to a specific dog breed.

Adolescent

Assay of specific IgE antibodies to disodium cromoglycate in serum from a patient with an immediate hypersensitivity reaction.

A 29-year-old man with pollen allergy had experienced immediate adverse reactions, such as itching of the eyes, rhinitis, wheezing, and general urticaria, after using disodium cromoglycate (DSCG) eye drops. The local symptoms were reproducible, and skin tests were strongly positive. With serum from the patient, a RAST was developed for the assay of IgE antibodies. The uptake on RAST disks was 6% of the total activity added, which was a significantly higher level than was found in sera from 35 randomly selected blood donors or in sera from 25 patients tolerating DSCG. By addition of DSCG to the patient's serum, 95% of the binding to paper disks could be inhibited. The induction of specific IgE antibodies was proposed to be a result of a combination of electrostatic and hydrophobic interaction of DSCG and a protein carrier. The substance would thus act as a hapten without any covalent binding to the carrier. DSCG may serve as a model for other nonreactive low-molecular-weight substances suspected to elicit type I-like adverse reactions.

Adult

Exposure to microorganisms, airway inflammatory changes and immune reactions in asymptomatic dairy farmers. Bronchoalveolar lavage evidence of macrophage activation and permeability changes in the airways.

Lung function, bronchial reactivity, serum antibody levels, bronchoalveolar lavage (BAL) fluid and the magnitude of exposure to airborne microorganisms during farm work were studied in 10 healthy farmers with no respiratory symptoms. Lung function and bronchial reactivity were not altered. An increase in the amount of antibodies against several microorganisms could be demonstrated but precipitating antibodies were observed only in two subjects. In the BAL fluid the nonsmoking farmers had elevated proportion of lymphocytes and elevated levels of albumin, fibronectin and angiotensin-converting enzyme while the levels of hyaluronan and procollagen III N-terminal peptide were within normal limits. The lung function, bronchial reactivity, serum antibody levels and BAL findings were not correlated with the exposure to airborne microorganisms. We conclude that healthy farmers, exposed to mold dust, may exhibit signs of alveolitis including albumin leakage, accumulation of inflammatory cells and alveolar macrophage activation.

Adult

Biological equilibration of allergen preparations: methodological aspects and reproducibility.

A method for biological equilibration (BE) of allergen reference preparations using the skin-prick test (SPT) method and histamine HCl 10 mg/ml as reference substance (reference method), was evaluated. The precision was low for weals less than 10 mm2. The slope (log weal area/log concentration) of allergen and histamine did not vary significantly between investigators and allergens. The median slopes were 0.39 (n = 384) and 0.34 (n = 397), for allergen and histamine, respectively (P less than 0.01). The concentration of allergen eliciting a weal of the same size as that of histamine HCl 1 mg/ml (Chl) in the median sensitive patient, 1000 Biological Units/ml (BU/ml), did not vary significantly between clinics/geographical regions (grasses, mites and moulds). As BE is repeatable between regions, BUs estimated by this method are generally valid. A high correlation (r = 0.91, P less than 0.001) was found between the median Chl as estimated with histamine 1 and 10 mg/ml as reference substance, respectively. Thus, this reference method for BE is valid. The precision of the SPT method with histamine HCl 1 mg/ml is not as good as with 10 mg/ml, which is therefore recommended as the reference concentration.

Adolescent

Results of biological standardization with standardized allergen preparations.

The aim of biological standardization (BS) is to equilibrate the activity (potency) of allergen extracts from different source materials. This was done by performing skin prick tests (SPT) on patients who were sensitive to one of the following 10 allergens: Birch, alder, hazel, timothy, rye grass, velvet grass, cultivated rye, mugwort, D. farinae and Cladosporium herbarum. Patient sensitivity varied within a range of three to four powers of ten for each allergen investigated. The weal size in each patient corresponding to that elicited by histamine 1 mg/ml was calculated using the model log (mean weal diameter) = a + b log (concentration). The correlation coefficients of the regression lines of the allergen dose response relationship were found to be greater than 0.85 in most cases. The median slope for all extracts was 0.24. The slope for Cladosporium was significantly steeper than that for pollens. The amount of material in microgram dry weight (d.w./ml) equal to 1000 biological units/ml (BU/ml) varied within a factor of three between species for all tested purified allergen preparations but Cladosporium. For Cladosporium, about 30 times more material was needed than for D. farinae. When using crude rather than purified material, it was necessary to use five to ten times more to elicit a reaction corresponding to 1000 BU/ml, but the difference was significant only for Cladosporium. The narrow range of allergen concentrations used by us as well as other investigators does not assure positive skin prick test results in all patients with clinical symptoms due to the allergen in question. Skin prick testing should therefore be done over a wide range of concentrations to improve the methods for BS.

Allergens