An animal model of hypersensitivity pneumonitis in the rabbit.

This study was devised to produce an animal model of hypersensitivity pneumonitis in order to study both the induction and the elicitation of the disease. Rabbits exposed by aerosol to large quantities of pigeon antigens developed a humoral, but not cellular, immunologic response. Moreover, their lungs were essentially normal histologically. A single i.v. injection of killed BCG in oil permitted the induction of pulmonary cell-medid hypersensitivity to the inhaled antigen, as well as the development of pulmonary lesions which were more severe than that caused by the administration of BCG alone. The humoral immunologic response to the inhaled antigen was not increased after BCG injection. Since many individuals are exposed to the etiologic agents of hypersensitivity pneumonitis for extended periods without developing the disease, these findings in animals suggest that some event may occur to induce cell mediated hypersensitivity in order to initiate the disease process. In addition, we have shown that animals with normal lung histology and circulating complement-fixing antibodies undergo serum complement (CH50) depression after an aerosol challenge with the specific antigen. Animals with circulating, complement-fixing antibodies, and inflamed lungs (BCG-induced failed to undergo a complement depression subsequent to an aerosol challenge with specific antigens. These results re consistent with those seen in symptomatic and asymptomatic pigeon breeders and suggest that antigen distribution through the lung is important in the pathogenesis of hypersensitivity pneumonitis.

Aspergillus fumigatus antigen induced eosinophilia in mice is abrogated by anti-IL-5 antibody.

A murine model of allergic bronchopulmonary aspergillosis (ABPA), developed by exposure to Aspergillus fumigatus antigens, demonstrated eosinophilia of peripheral blood (PB), bone marrow (BM), and lung. The eosinophilia was abrogated by monoclonal anti-interleukin-5 (IL-5) antibody (TRFK-5) and not by an isotype control antibody (GL 113). Eosinophils in PB were enumerated from stained smears and their relative increase or decrease in cells from BM and lung was determined by an eosinophil peroxidase (EPO) assay (measured in optical density). Intraperitoneal injection of TRFK-5 in mice exposed to A. fumigatus antigen produced a significant reduction in eosinophils (PB 6.6 +/- 1.14% vs. 3.8 +/- 0.8%, P < .01) and EPO production in BM (0.935 +/- 0.03 vs. 0.615 +/- 0.02, P < .001). A similar reduction in EPO production in the lung (0.691 +/- 0.12 vs. 0.495 +/- 0.05, not significant) was also reflected in the histopathology for the different groups of mice. These findings confirming the role of IL-5 in eosinophilia, although not surprising, are significant in elucidating the immunopathogenesis of ABPA in the murine model. We conclude that in this model, eosinophilia may be due largely to the Th2 cytokine -IL-5 induced by A. fumigatus antigens.

Hyperimmune serum modulates allergic response to spores in a murine model of allergic aspergillosis.

Allergic bronchopulmonary aspergillosis (ABPA) is a disease in atopic asthmatics characterized by eosinophilia and elevated levels of immunoglobulin E (IgE) and IgG antibodies to the ubiquitous fungus Aspergillus fumigatus (Af). The role of specific antibodies in the disease process is not clear. In this study, BALB/c mice were injected with hyperimmune serum from syngeneic mice exposed to soluble antigen of Af. These mice were then exposed to either Af spores or soluble antigen. Total IgE, Af-specific IgG1 (enzyme-linked immunosorbent assay) in serum, and eosinophils (eosinophil peroxidase assay) in lungs and bone marrow were measured. Histologic sections of lungs were examined for cellular infiltration and morphologic changes. Results indicate a suppression of increase in levels of antibodies and eosinophilia in mice receiving immune serum and exposed to spores compared with controls receiving phosphate-buffered saline treatment. Spores being the primary source of exposure to Af in ABPA, these results are significant in understanding the role of preexisting specific antibodies in patients.

Unique and shared IgE epitopes of Hev b 1 and Hev b 3 in latex allergy.

Of the several latex proteins cloned and expressed, the rubber elongation factor, Hev b 1, and the closely related Hev b 3, represent two major allergens associated with latex allergy. Although both allergens demonstrated IgE binding with sera from latex allergic patients, it was not known whether these two molecules shared any epitopes. Hence, in the present study using health care workers (HCW) and spina bifida (SB) patients with latex allergy, we investigated the IgE binding epitopes in Hev b 1 and Hev b 3. Recombinant Hev b 1 and Hev b 3 were expressed in a prokaryotic expression system, while overlapping decapeptides of Hev b 1 and Hev b 3 were synthesized on derivatized cellulose membrane. Eight IgE binding epitopes for Hev b 1 and eleven for Hev b 3 were identified using sera from latex allergic patients with SB. On further analysis of synthetic peptides encompassing these epitopes, similar IgE antibody reactivity was demonstrated with three Hev b 1 epitopes b1E3, b1E5, b1E6 and two Hev b 3 epitopes; b3E10 and b3E 11. For Hev b 1, a unique IgE binding epitope was identified in the region of amino acid residues 16-25. In competitive ELISA, peptides bIE2 and bIE4 together inhibited 58% of IgE binding of Hev b 1, while b3E5 showed 22% inhibition in the IgE binding of Hev b 3. The results of the present study suggest that the understanding of linear and conformational IgE epitopes in the major latex allergens may provide better insight into the structure-function relationship of the allergens, and may lead to the development of better patient care and management strategies in latex allergy.

Study of two controlled-release theophylline preparations.

Two sustained-release theophylline preparations, one a 12-hour and the other a 24-hour product, were compared in 20 subjects with stable asthma. All patients had good control of symptoms, and the calculated pharmacokinetic parameters were comparable for both drugs. The 24-hour sustained-release preparation can thus be utilized in the therapy of asthma.

Pigeons breeders' disease. I. Serum immunoglobulin concentrations; IgG, IgM, IgA and IgE antibodies against pigeon serum.

The clinical course of a patient with severe acute and chronic hypersensitivity pneumonitis of the pigeon breeders' disease (PBD) type was evaluated. A significant remission of symptomatic, roentgenographic and pulmonary function abnormalities occurred after cessation of exposure to antigen. Immunoglobulin G (IgG), A (IgA) and M (IgM) antibody activity against pigeon serum was demonstrated in the patient's serum by a solid phase radioimmunoassay (RIA) technic. The levels of these antibodies fell subsequent to elimination of antigen inhalation. The RIA technic was used to demonstrate IgG, IgA and IgM antibody activity in single serum samples of four other patients with PBD, and this technic may have diagnostic value. Although no immunoglobulin E (IgE) antibody was demonstrable by RIA, a heat-liable, reaginic antibody was also detectable in the serum of the primary case. In addition, Arthus type cutaneous reactivity was passively transferred to the skin of a volunteer subject using heated serum from a patient with PBD.

Early detection of hypersensitivity pneumonitis in office workers.

Symptoms of hypersensitivity pneumonitis in three employees in an office building led to an investigation of their work environment. An open spray water air cooling system was implicated when inhalation challenge with the spray water caused acute illness in one of them. A questionnaire survey of the 4,023 co-workers identified 48 other suspect cases, and laboaratory studies confirmed hypersensitivity pneumonitis in three additional workers of this group. A significant change in pulmonary function, occurring only after exposure to the work environment, was the most useful laboratory finding and was found in five workers with no other pulmonary abnormalities, but not is asymptomatic workers or controls, since five of the six patients with hypersensitivy pneumonitis worked in offices cooled by the spray water system and since three had positive responses to inhalation challenge, use of the spray water system was discontinued. The affected workers improved after they were removed from the office complex.

Childhood hypersensitivity pneumonitis due to dove antigens.

A case of hypersensitivity pneumonitis due to doves is reported and compared with other cases due to dove or pigeon antigens reported in children. The diagnosis is substantiated by the presence of precipitating antibody to dove and pigeon serum, clinical improvement after contact with the doves was broken, and a positive response to inhalation challenge with pigeon serum. The insidious nature of this disease is emphasized as well as the importance of having detailed environmental information in children with unexplained respiratory disease.

Serum antibodies and their role in antibody-dependent cell-mediated cytotoxicity in aspergillosis.

A total of 22 sera from patients with aspergilloma and allergic bronchopulmonary aspergillosis (ABPA) were examined concomitantly for specific antibody against Aspergillus fumigatus antigen and for their activity in antibody-dependent cell-mediated cytotoxicity (ADCC) against Aspergillus antigen-coated target cells. These sera demonstrated significant precipitin bands in agar gel double diffusion test (78% of ABPA and 75% of aspergilloma sera), while in indirect immunofluorescence studies all sera showed positive reactivity with a titer distribution of 1:40 to 1:160 and 1:40 to 1:320, respectively, for ABPA and aspergilloma sera. In enzyme-linked immunosorbent assay all sera demonstrated titers varying from 1:200 to 1:6400. Several sera also displayed marked cytotoxic reactions against A. fumigatus antigen-coated SB target cells in ADCC assays using normal lymphocytes as effector cells (35% of aspergilloma and 25% of ABPA sera). These findings suggest a role for ADCC activity in patients with Aspergillus infections.

Allergic bronchopulmonary aspergillosis.

Allergic bronchopulmonary aspergillosis (ABPA) occurs as a complication of asthma. It presents as an infiltrative pulmonary disorder with respiratory and systemic symptoms, eosinophilia, elevated total serum IgE, and skin sensitizing, as well as precipitating antibodies to Aspergillus fumigatus. Sputum cultures are not always positive for the organism. If unchecked, the disease may lead to bronchiectasis and ultimately pulmonary fibrosis. Therapy consists mainly of corticosteroids.

Hypersensitivity studies in popple (Aspen tree) peelers.

In the course of investigation of the relationship between hypersensitivity pneumonitis and the wood industry 45 popple peelers were studied. Four roentgenograms consistent with diffuse lung disease were found. All cases were in examinees who were also dairy farmers. Questionnaires revealed no instance of clinical symptoms of hypersensitivity pneumonitis. Gel immunodiffusion studies were positive to various antigens in eight cases, but no correlation could be made with pulmonary function or chest roentgenogram abnormalities in relation to occupation. Cultures of the wood and the surrounding environment yielded thermophilic fungi as well as other known antigenic material. It is concluded that the occupation of popple peeling does not represent an increased risk of development of hypersensitivity pneumonitis.

Serum angiotensin-converting enzyme levels in patients with pigeon-breeder's disease.

The serum concentration of angiotensin-converting enzyme is frequently elevated in individuals with active sarcoidosis. The enzyme is presumably actively synthesized by the epithelioid and giant cells of the granuloma. Hypersensitivity pneumonitis, resulting from the inhalation of antigens from pigeons by susceptible individuals, is associated with the development of a granulomatous interstitial and alveolar infiltrate in the pulmonary parenchyma. Because the clinical and pathologic presentation may mimic that of sarcoidosis, we compared the serum levels of angiotensin-converting enzyme in these two diseases. The concentration of angiotensin-converting enzyme is not elevated in individuals with active hypersensitivity pneumonitis, in contrast to its frequent elevation in sarcoidosis. We suggest that the granulomatous response in hypersensitivity pneumonitis may differ at a biochemical level from that of sarcoidosis, since the synthesis of angiotensin-converting enzyme does not appear to be increased.

Antigen-induced enhancement of bronchial reactivity.

Bronchial hyperreactivity as determined by the airway response to methacholine was evaluated pre- and post-antigen challenge in three patients with specific antigen sensitivity. No significant change in pulmonary function was noticed after inhalation of antigen alone. However, transient but significant increase in methacholine responsiveness followed the antigen challenge. The studies indicate a need for a nonspecific inhalation challenge following a negative antigen challenge when clinically indicated.

Immune responses to Aspergillus fumigatus and Pseudomonas aeruginosa antigens in cystic fibrosis and allergic bronchopulmonary aspergillosis.

Allergic bronchopulmonary aspergillosis (ABPA) in Cystic Fibrosis (CF) is well documented. Aspergillus fumigatus is the causative agent of ABPA, and Pseudomonas aeruginosa particularly the mucoid variety has been frequently isolated from the sputum of patients with CF. This study investigates the cellular and humoral immune response to both A fumigatus and P aeruginosa antigens in patients with CF and ABPA (CF/ABPA), CF only, and healthy controls. The A fumigatus and P aeruginosa antigen specific IgE and IgG in sera and peripheral blood mononuclear cell culture supernatants (PBMC sups), lymphoproliferation to antigens, and leukotriene B4 (LTB4) were measured. Results indicate significant elevated levels of A fumigatus specific IgG (A fumigatus-IgG) and Paeruginosa-IgE in serum. Significant Paeruginosa-IgG was measured in PBMC sups. The concanavalin A nonbinding A fumigatus antigen, previously shown to induce specific T-cell responses in vitro in patients with ABPA, elicited significant lymphoproliferative response in a greater proportion of patients with CF/ABPA and not in CF or controls, underlining the importance of this antigen in the diagnosis of ABPA. In contrast, a greater proportion of the CF group responded to P aeruginosa antigens compared with the controls and CF/ABPA. Hence, the CF and CF/ABPA groups respond to both P aeruginosa and A fumigatus antigens with the former group responding strongly to P aeruginosa and the latter to A fumigatus antigens.

Sarcoidosis in hypersensitivity pneumonitis.

An abnormal chest x-ray film showing hilar adenopathy, diffuse interstitial pulmonary infiltrations, or both, combined with a tissue biopsy revealing noncaseating granuloma, are suggestive of sarcoidosis; however, non-caseating granuloma may also be found in other forms of pulmonary disease. Immunologic and environmental evaluation of three patients with the diagnosis of sarcoidosis made by the above criteria, revealed hypersensitivity pneumonitis in all. Since therapeutic considerations in these two diseases are different (avoidance being the mainstay in hypersensitivity pneumonitis), all methods to ensure a correct diagnosis should be employed.

Evaluation of methods to detect antibodies against Aspergillus fumigatus.

Seventy-five sera, including sera from 23 patients with aspergillosis, 17 with tuberculosis, asthma or carcinoma, and 35 normal controls, were studied for antibody activity against Aspergillus fumigatus by agar gel double diffusion (DD), counterimmunoelectrophoresis (CIE), indirect hemagglutination (IHA) and indirect immunofluorescence (IF). Metabolic antigens produced from a selected strain of A. fumigatus were used for DD, CIE and IHA, while slide cultures of the same organism were used for IF. The results indicated that sera from the patients with aspergillosis had high IHA and IF titers, while only 91 and 87% were positive for CIE and DD methods, respectively. IHA was more sensitive than DD and CIE, and IF was equally sensitive but had false-positive reactions. CIE was simple and fast, but had false-negative and occasionally false-positive results.

Aspergillus fumigatus peptides differentially express Th1 and Th2 cytokines.

Relevant allergens from Aspergillus fumigatus associated with allergic bronchopulmonary aspergillosis (ABPA) have been cloned and expressed. The pathogenesis of ABPA probably depends on specific cytokines and immunoglobulins secreted by lymphocytes on stimulation with different epitopes of those allergens. In the present study, we synthesized peptides of 12-16 amino acids from the sequence of Asp fI and compared their immunological responses in four mice strains (BALB/c, C57BL/6, AKR, and CBA). Of the five peptides studied for their cytokine profile, one showed a clear Th1, whereas another showed a Th2 response. The remaining three peptides varied in their immunoreactivity. The results suggest that a number of epitopes of diverse activities are present in individual molecules and may be involved in the pathogenesis of ABPA through differential cytokine secretions.

Immunodominant peptide epitopes of allergen, Asp f 1 from the fungus Aspergillus fumigatus.

Aspergillus fumigatus ribotoxin Asp f 1 is a major allergen with IgE binding activity to serum of a majority of patients with allergic bronchopulmonary aspergillosis (ABPA). The IgE binding epitopes or the T-cell stimulatory peptides of this molecule have not been studied. In the present investigation, we have synthesized linear decapeptides spanning the whole molecule of Asp f 1 and analyzed their IgE binding properties. We have also synthesized peptides based on their possible T-cell stimulatory properties and studied the stimulation of peripheral blood mononuclear cells from ABPA patients and normal controls. Several peptides demonstrated distinct IgE antibody binding response against sera from ABPA patients and proliferative response against peripheral blood mononuclear cells from the patients. From the results, it can be concluded that the carboxy-terminal region of Asp f 1 representing amino acid residues 115-149 involved in both humoral and cell mediated immunoresponses in ABPA.

Hypersensitivity pneumonitis.

Hypersensitivity pneumonitis is a syndrome caused by sensitization to any of a wide variety of environmental antigens and results in a pulmonary immunologic inflammatory process. The clinical syndrome occurs in a temporal relationship to exposure to the offending agent with resultant systemic or respiratory symptoms with defined chest radiographic and pulmonary function abnormalities. Definitive immune responses include those of the humoral and cellular system. Avoidance of exposure as treatment results in reversal of the clinical abnormalities.

Surgery for sinusitis and aspirin triad.

An 11-year retrospective study was conducted to evaluate the surgical treatment of sinusitis in aspirin-triad patients. Twenty-five patients were selected for surgery when their sinus disease and asthma progressed despite intense medical treatment. Sixteen patients had radiographic evidence of severe ethmoid disease, and their initial surgical procedures were limited (i.e., bilateral intranasal ethmoidectomies). Of these 16 patients, 6 required subsequent surgery for recurrent sinusitis. Nine of the 25 patients had radiographic evidence of severe antral, as well as ethmoid, disease. Their initial surgical procedures were radical, i.e., bilateral Caldwell-Luc operations with intranasal and transantral sphenoethmoidectomies. None of these 9 patients required further surgical treatment for control. This review indicates that if antral and sphenoid disease are detected in association with ethmoid involvement, a radical surgical approach is the operation of choice.