Respiratory health effects and immunological response to Thermoactinomyces among sugar cane workers in Nicaragua.

Specific sensitization and respiratory effects associated with the inhalation of sugar cane dust were evaluated in a group of 51 Nicaraguan workers exposed to bagasse. A questionnaire interview, lung function test, serum precipitin tests for Thermoactinomyces sacchari and T. vulgaris, and immunoglobulin E tests for specific environmental allergens were performed for each worker. Twenty-one workers reported at least one respiratory symptom and 16 reported possible symptoms of bagassosis. Six workers demonstrated acute symptoms, 1 had chronic symptoms, and 9 had the reacutized form of the disease. A higher proportion of precipitin response to T. sacchari and T. vulgaris was found in workers reporting symptoms suggestive of acute bagassosis. A possible restrictive ventilatory pattern was observed in 8 subjects and a mild airway obstruction in 1 subject. Priority must be given to a surveillance and exposure prevention program for workers employed in sugar cane production and processing.

[A case of hypersensitity pneumonitis due to the contamination of a dehumidifier by Thermoactinomyces vulgaris].

A 22-year-old man was admitted to our hospital because of high fever and shortness of breath. A chest CT showed centrilobular nodules and ground-glass attenuation in both lungs. Transbronchial lung biopsy specimens showed alveolitis including the infiltration of mononuclear cells and granulomas. He responded to treatment, but upon returning home the radiological findings and clinical symptoms reappeared. We focused on a dehumidifier because it had been used continuously in his room. Thermoactinomyces vulgaris presented in the air filter of the dehumidifier. The test of precipitation in response to Thermoactinomyces vulgaris was positive. The condition inside the dehumidifier of a high temperature and high humidity were suitable for the proliferation of Thermoactinomyces vulgaris. We diagnosed hypersensitity pneumonitis due to a dehumidifier contaminated by Thermoactinomyces vulgaris.

Specific IgG to Thermoactynomices vulgaris, Micropolyspora faeni and Aspergillus fumigatus in building workers exposed to esparto grass (plasterers) and in patients with esparto-induced hypersensitivity pneumonitis.

BACKGROUND: Esparto is one the most frequent causes of hypersensitivity pneumonitis in Spain. OBJECTIVE: Determination of risk factors in the working environment that could explain the sensitisation process, and assessment of the differences in specific IgG levels to Aspergillus fumigatus, Saccharopolyspora rectivirgula and Thermoactynomices vulgaris in patients with hypersensitivity pneumonitis induced by esparto, exposed healthy plasterers and control population. METHODS: Determination of precipitins and specific IgG to Aspergillusfumigatus, Saccharopolyspora rectivirgula and Thermoactynomices vulgaris in the three previously mentioned groups. Questionnaire on possible risk occupational and extra-occupational factors. RESULTS: Both healthy and exposed plasterers have higher levels of specific IgG to Aspergillus fumigatus, Saccharopolyspora rectivirgula and Thermoactynomices vulgaris than the healthy controls. The patients had higher levels of IgG than exposed healthy plasterers only to Thermoactynomices vulgaris. Precipitins were detected in only two patients. There were no occupational factors influencing on the sensitisation process. CONCLUSIONS: Specific IgG is an occupational exposure marker among plasterers, but it has not been possible to establish a cut off point that differentiates exposed subjects from affected ones. This determination has a greater sensitivity than precipitins. We did not identify occupational or extra-occupational risk factors that facilitate the sensitisation process.

Immunochemical studies of a purified antigen from Micropolyspora faeni.

A pure antigen fraction was isolated from the crude culture filtrate of Micropolyspora faeni by gel filtration and affinity chromatography. The isolated antigen has a mol. wt of approximately 16,000 and an isoelectric point of pH 3.8. The major amino acid content of this fraction includes glycine, glutamic acid, aspartic acid and alanine. This antigen fraction reacted with the sera of all 15 farmer's lung patients and 20 asymptomatic farmers with circulating anti-M. faeni antibodies. An ELISA method was developed using the purified antigen to detect specific circulating antibodies against M. faeni in farmer's lung patients.

Angiotensin-converting enzyme in farmer's lung.

Serum levels of angiotensin-converting enzyme were measured in a group of patients with farmer's lung and a group of precipitin-positive subjects with no history of farmer's lung. The levels did not differ significantly from control subjects matched for age and smoking history. The mean serum level of angiotensin-converting enzyme in a group of acutely ill patients with farmer's lung was significantly reduced. An acute challenge of three patients with Micropolyspora faeni did not produce an increase in serum levels of angiotensin-converting enzyme. These studies suggest that an increased serum level of angiotensin-converting enzyme can be a diagnostic aid in making a differential diagnosis between sarcoidosis and farmer's lung.

Usefulness of skin test in Farmer's lung.

The skin test with hay extract or with Micropolyspora faeni (MF), is not commonly used in the diagnosis of Farmer's lung (FL), as it is not considered specific. In our study, we have applied the intracutaneous test with these antigens in 26 patients affected with FL; 18 of them were still in contact with the antigen and the remaining eight had not been in contact with the hay during the previous year. Twenty-five asymptomatic farmers (AF) served as a control group. In the first group and with hay extract, the immediate reading (I) was positive in 15 of 18 (83.3 percent), the late reaction (L) in 18 of 18 (100 percent), and the delayed one (D) in eight of 18 (44.4 percent). In the 25 AF, the results were as follows: I, seven of 85 (28 percent); L, 17 of 25 (68 percent); and D, one of 25 (4 percent). Consequently, the differences between both groups were significant: I, p less than 0.01; L, p less than 0.05; and D, p less than 0.01. Using MF as an antigen, the test is somewhat less effective: p less than 0.02, p less than 0.02, and p less than 0.2, respectively. These results suggest that the intradermal injection with hay extract is an easy, effective test in the diagnosis of FL, and at the same time, a better means of distinguishing FL patients from AF than the precipitation test.

The absence of a relationship between serum precipitins and pulmonary disease in a community.

To study the role of serum precipitins in respiratory illness in a community, sera obtained from 3,047 residents of Tucson, Ariz., were tested for the presence of precipitating antibody to a battery of antigens. Positive reactions were obtained in 54 subjects (1.8 percent), a lower incidence than has been reported previously. The majority of these subjects were older than 54 years of age, an age distribution significantly older than the entire sample (P less than 0.01). Pulmonary function among the subjects with positive precipitin reactions was not significantly different from that of the asymptomatic nonsmokers of the entire sample. None of the subjects who were lifelong residents of Arizona had serum precipitins to any of the thermophilic actinomycetes antigens which were used in the testing. These antigens have been found in association with extrinsic allergic alveolitis, most frequently among patients living in the north central United States and were derived from strains of Micropolyspora faeni, Thermoactinomycetes candidus and vulgaris. Each subject with precipitins to one or more of the tested antigens was matched by age, sex, and socioeconomic class with two subjects from the sample who had negative precipitin reactions. The groups did not differ in their prevalence of respiratory symptoms or abnormalities of pulmonary function. We conclude that the presence of precipitating serum antibodies among subjects in a community is not indicative of the presence of immunologic pulmonary disease but merely reflects previous exposure to the tested antigen. In addition, individuals whose sera contain precipitating antibody appear to have no increased tendency to develop other types of pulmonary disease.

Familial aggregation of IgG antibody response to antigens associated with farmer's lung.

The levels of circulating IgG antibodies to Aspergillus umbrosus, Aspergillus fumigatus, Thermoactinomyces vulgaris, and Micropolyspora faeni were determined by enzyme immunoassay in 197 subjects selected for a study of farmer's lung (FL). The material consisted of five study groups: 37 patients with clinically confirmed FL, 31 spouses of the patients, 44 immediate relatives of the patients, 35 immediate relatives of the patients' spouses, and 50 unrelated people who were spouses of the 79 people in both relative groups. The mean titres of IgG antibodies to all four microbes were highest in patients with clinically established FL. In the other groups the mean titre of Aspergillus umbrosus, a mould found much more frequently in Finnish farm environments than other moulds under study, was significantly higher (p less than 0.01) in the relatives of FL patients than in other people. This finding remained irrespective of whether the subjects had suffered from FL symptoms or not or whether they worked or lived on the same farm as the patient or on a different one. The difference in the mean titre was not due to the differences between the study groups in age, sex, smoking habits, atopic background, frequency of handling of plant materials, or time interval from the most recent handling of visibly mouldy hay. The results imply that genetic factors may be important in the IgG antibody response to microbial antigens associated with FL.

Use of a serologically distinct strain of Thermoactinomyces vulgaris in the diagnosis of farmer's lung disease.

During investigations into farmer's lung disease it was noticed that Micropolyspora faeni (MF) was isolated and identified with ease, but difficulty was encountered in the identification of Thermoactinomyces vulgaris (TV), although an organism resembling TV was frequently isolated. Extracts prepared from the isolates resembling TV (called Thermoactinomyces vulgaris variant, TVV) when tested against the standard TV antiserum by double diffusion, did not produce any precipitin lines. When TVV extracts were tested against the serum of a patient from whom TVV had been isolated from the sputum, +++ precipitin lines were observed. With the TVV extracts the authors have demonstrated the existence of a serologically distinct strain of TV, and also that it would be a useful addition to the routine testing of antibodies to farmer's lung disease.

Micropolyspora faeni causes airway inflammation but not hyperresponsiveness in sensitized ponies.

We assessed the effect of aerosol Micropolyspora faeni challenge in two groups of ponies by measuring lung function, airway reactivity to aerosol histamine, and bronchoalveolar lavage fluid cytology. One group of ponies was sensitized by subcutaneous injection of M. faeni in complete Freund's adjuvant, and the other group served as control. In both groups of ponies, measurements were made at base line and 5 h after aerosol administration of 30 ml of saline or 30 ml of 1% wt/vol particulate M. faeni antigen in saline. Saline challenge had no effect on any of the measured variables. M. faeni challenge had no effect on pulmonary mechanics or gas exchange in the control group but significantly increased respiratory frequency and minute ventilation and decreased arterial CO2 tension in the sensitized ponies. In both groups of ponies, aerosol M. faeni challenge significantly increased total white blood cell count and neutrophil numbers in bronchoalveolar lavage fluid while large mononuclear cell numbers decreased. Airway responsiveness was unaltered by saline or M. faeni challenge in both pony groups. We conclude that aerosol M. faeni challenge induces pulmonary neutrophilia and abnormalities of ventilation but is not accompanied by airway hyperresponsiveness in sensitized ponies.

Local and systemic antibody production in horses affected with chronic obstructive pulmonary disease.

An enzyme-linked immunosorbent assay (ELISA) was used to quantify isotype-specific antibody to Micropolyspora faeni and to Aspergillus fumigatus in the sera and bronchoalveolar lavage fluid (BALF) of normal horses, horses with chronic obstructive pulmonary disease (COPD) and horses with other chronic respiratory diseases. Elevated antibody levels were not detected in the sera of affected horses. However, both IgE and IgA antibody to both allergens was significantly elevated in BALF in COPD affected horses sampled both when symptomatic and asymptomatic. Elevated levels were also found in animals that developed a chronic cough after an acute onset with symptoms compatible with a respiratory virus infection. In one animal a ten fold increase in IgE antibody to the two allergens developed after an interval of 7 weeks. These findings are supportive of a central role of local IgE antibody to mould allergens in the immunopathogenesis of COPD, and also suggest that respiratory viral infection may predispose to the development of COPD in some horses.

The effects of sodium cromoglycate on antigen inhalation challenge in two horses affected with chronic obstructive pulmonary disease (COPD).

80 mg sodium cromoglycate (SCG) was administered by inhalation to two COPD-affected animals known to have respiratory hypersensitivity to Micropolyspora faeni. SCG treatment 20-30 minutes prior to inhalation challenge with M. faeni prevented exacerbation of respiratory disease, usually seen 4-8 hours after challenge. duration of protection against antigen challenge after a single SCG treatment was 4-5 days. The duration of protection was not prolonged by reducing the frequency of antigen challenge. Multiple antigen challenge, using M. faeni and Aspergillus fumigatus, shortened the protective period of SCG to 3 days.

Quantitation and localization of immunoglobulin E containing cells in bovine lymphoid tissues.

Although recent studies have begun to describe and quantify IgE responses in bovine serum and secretions, little is known about the distribution and quantity of IgE containing cells in cattle. In the present study, cells with cytoplasmic IgE were quantitated in bovine lymphoid tissues, using immunoperoxidase staining and evaluation by an image analysing computer (Quantimet). Frozen sections from retropharyngeal, bronchial and mesenteric lymph nodes, tonsil and spleen were stained from 11 calves, some of which had been exposed to antigen by aerosol or injection. Although individual variability was considerable, bronchial and mesenteric lymph nodes generally contained the greatest percentage of IgE containing cells, while retropharyngeal lymph node, tonsil, and spleen had less. Parenteral immunization with ovalbumin appeared to increase the splenic percentage, while aerosol exposure to ovalbumin was associated with a greater percentage of IgE containing cells in bronchial lymph nodes. Comparison of the present results with those reported for other species shows some similar trends in IgE localization.

A study on the possible role of chymotrypsin in the aetiology of equine chronic obstructive pulmonary disease (COPD).

The chymotrypsin activity of seven batches of Micropolyspora faeni and of five batches of Aspergillus fumigatus culture extracts, prepared for inhalation challenge in horses, was assayed and was found to range between 0.29 and 1.45 units/mg protein and 0.02 and 0.20 units/mg protein respectively. Horses affected with chronic obstructive pulmonary disease (COPD) were challenged with two batches of each antigen which had different chymotrypsin activities and no significant correlations were found between the degree of response to challenge and the chymotrypsin activity of the antigens. Inhalation of two doses of nebulised, purified chymotrypsin over 4 days did not induce signs of respiratory disease in COPD-affected horses. However, repeated chymotrypsin inhalations after an interval of 3 weeks caused an exacerbation of signs of COPD in one horse. These studies suggest that, although repeated inhalation of purified chymotrypsin may induce respiratory hypersensitivity in horses, the chymotrypsin-like enzymes of M. faeni and A. fumigatus do not play a major role in the precipitation of clinical signs of equine COPD.

Antioxidant therapy partially blocks immune-induced lung fibrosis.

A mouse model of hypersensitivity pneumonitis was generated by challenge with a thermophilic actinomycete. Oxygen radical scavengers were administered to challenged mice: vitamin E at 1000 units daily, polyethylene glycol-superoxide dismutase (SOD) at 500 units daily, polyethylene glycol-catalase at 10,000 units daily, 1,3,dimethyl-2-thiourea (DMTU) at 2 mg daily, and the biomimetic SOD, copper(II) [diisopropyl salicylate]2 (CuDIPS) at 1 mg daily. At three weeks after actinomycete challenge, a 10-fold increase in bronchoalveolar (BAL) cell number was observed. Treatments with catalase or DMTU were without effect on the BAL cell number in challenged mice. However, infusion of vitamin E was associated with an increased BAL cell influx (15-fold increase at two and three weeks). Similarly, treatment with PEG-SOD and CuDIPS resulted in an increase in cell number at two and three weeks. PEG-SOD or CuDIPS treatment resulted in a strong neutrophilia, whereas control challenged mice had a cellular influx mostly of macrophages and lymphocytes. Vitamin E treatment of challenged mice led to an increased T lymphocyte recruitment at two and three weeks. In vitro studies showed that actinomycete challenge was associated with an enhancement of alveolar macrophage O2- release, which was blocked by PEG-SOD, vitamin E, or DSC treatment but was unaffected by catalase or DMTU treatment. In control challenged mice, there was a 25-fold increase in the BAL albumin concentration at two weeks. PEG-SOD, vitamin E, or CuDIPS treatment all decreased the albumin concentration; the three modulators also diminished lung fibrosis at two or three weeks, as seen by a decrease in lung hydroxyproline and collagen synthesis by lung fibroblasts. Examination of sections from lungs of challenged animals showed evidence of cellular infiltrates around the bronchi and the blood vessels. Challenged mice given continuous infusions of vitamin E, SOD, or CuDIPS had lung histological scores that were significantly lower than control challenged mice or challenged mice treated with catalase or DMTU. Thus, therapies based on O2- scavenging or treatment with a general antioxidant such as vitamin E may hold some promise in the treatment of hypersensitivity pneumonitis.

Effect of inhalation of organic dust-derived microbial agents on the pulmonary phagocytic oxidative metabolism of guinea pigs.

The effect of inhalation exposure of various biological agents associated with organic dusts on the function of guinea pigs pulmonary phagocytes was investigated. Agents included antigens of Erwinia herbicola, Thermoactinomyces vulgaris, and Aspergillus fumigatus; endotoxin of Erwinia herbicola; bacterial protease; and a fungal glucan preparation. Pulmonary parameters monitored in this study were cellular differential counts from bronchoalveolar lavage, and superoxide anion and/or hydrogen peroxide production by phagocytic cells. Most of the agents caused an influx of neutrophils, lymphocytes, and red blood cells to the lung, and an enhancement of secretion of reactive oxygen species by pulmonary phagocytes. However, the relative magnitude of the inflammatory response varied greatly among these biological agents. In general, antigens of Erwinia herbicola and Aspergillus fumigatus were most potent, while bacterial protease was least effective.

Detection of antigens in lung biopsies by immunoperoxidase staining in extrinsic allergic bronchiolo-alveolitis (EABA).

Open lung biopsy specimens of 14 patients with extrinsic allergic bronchiolo-alveolitis (EABA) were studied by immunoperoxidase staining with avidin-biotin-complex method using nonconjugated antisera to Micropolyspora faeni, Thermoactinomyces vulgaris, Aspergillus fumigatus, and Aspergillus umbrosus. Antigenic intracellular material was found in macrophages and giant cells of granulomas in the lung tissue of 8 patients. This finding supports the hypothesis which suggests that macrophages and T-lymphocytes contribute to the tissue injury in EABA.

In vitro response of lymphocytes to Micropolyspora faeni extract in cattle.

The in vitro stimulation of small lymphocytes to blast formation, measured by the rate of 3H-thymidine incorporation, was used to study the occurrence of cells sensitive to antigens of Micropolyspora faeni in cattle. M faeni extract induced a significant stimulation index in lymphocytes from the peripheral blood cells of cattle from an endemic area in autumn but rarely in spring. Blood lymphocytes from animals from a non-endemic area tested during the winter period rarely showed a positive reaction or only a relatively weak one. On the other hand, lymph node cells, particularly from bronchial lymph nodes, showed positive results in all investigated animals and even in those from non-endemic areas. In three-months-old calves, positive results were obtained mainly with cells from bronchial lymph nodes. It seems therfore that sensitisation to M faeni antigen is a widespread phenomenon but additional circumstances seem to be required for the clinical manifestation of farmer's lung disease in cattle. The most important factor is probably strong and repeated exposure to the M faeni organism. Whether or not existing reactive lymphocytes against M faeni antigen are directly involved in the pathogenesis of farmer's lung disease in cattle by producing a delayed type reaction remains to be clarified.

Chronic obstructive pulmonary disease (COPD) in horses: aetiological studies: responses to intradermal and inhalation antigenic challenge.

Micropolyspora faeni and Aspergillus fumigatus were identified as common causes of respiratory hypersensitivity in horses affected with chronic obstructive pulmonary disease (COPD). Rye grass pollen and an Actinomycete evoked respiratory allergy in a few horses. Not infrequently, individual horses were found to have respiratory hypersensitivity to two or more antigens. The methods used to examine for allergy were intradermal testing and inhalation challenge with environmental antigens. An intradermal test using an M faeni extract was demonstrated to be suitable for diagnostic use in horses previously accurately diagnosed as suffering from COPD. In contrast, the A fumigatus antigen used proved unsatisfactory for such a purpose. Skin reaction to M faeni and A fumigatus extracts by horses affected with COPD indicated that the hypersensitivity was a dual one--a weak response shortly after injection followed by an Arthus-like response 4 to 8 hours later. As a parameter for monitoring responses to inhalation challenge, maximum intrathoracic pressure change (max delta Ppl) proved satisfactory, whereas changes in partial pressure of arterial oxygen (PaO2) did not.

The presence of precipitating antibodies in the sera of horses with chronic obstructive pulmonary disease (COPD).

The sera of horses affected and not affected with chronic obstructive pulmonary disease (COPD) were examined for precipitins to Micropolyspora faeni and Aspergillus fumigatus. Precipitins to both antigens were not restricted to COPD cases but occurred more frequently in animals affected with COPD. Many animals without detectable precipitins responded clinically to inhalation challenge with these antigens.