Different classes of antibody activities to Trichosporon cutaneum antigen in summer-type hypersensitivity pneumonitis by enzyme-linked immunosorbent assay

To assess the diagnostic value of antibody activities to Trichosporon cutaneum antigen in patients with summer-type hypersensitivity pneumonitis in Japan, we measured the serum IgG, IgA, and IgM antibodies to saline-soluble antigen of homogenized T. cutaneum cells by enzyme-linked immunosorbent assay (ELISA). The IgG antibody activity was observed in serum samples from 25 (83%) of the 30 patients and in 9 (82%) of the 11 asymptomatic family members. The IgA antibody activity was found in 25 (83%) of the patients but in only 3 (27%) of asymptomatic family members. Unexpectedly, we found no significant correlation between the IgG and IgA antibody activities to the same antigen in individual patients. Moreover, although the IgG antibody activity was related to the precipitating antibody, the IgA antibody activity was not. The IgM antibody activity by ELISA was found in only 5 (17%) of the patients. The IgG+IgA+IgM antibody activity was demonstrated in 27 (90%) of the 30 patients, and of 3 serum samples that were found negative by the ELISA method, 2 were positive by the IFA method. We conclude that the detection of the serum IgG and IgA antibody activities to T. cutaneum antigen by ELISA is a simple and useful serodiagnostic test for summer-type hypersensitivity pneumonitis. Measurement of the IgA antibody activity is especially helpful in discriminating symptomatic from asymptomatic family members.     

C1q and C3 in bronchoalveolar lavage fluid from patients with summer-type hypersensitivity pneumonitis

Immune complexes have been thought to participate in the pathogenesis of hypersensitivity pneumonitis, but the role of complement components is not defined. In our study of nine patients with summer-type hypersensitivity pneumonitis (summer-type HP), C1q in bronchoalveolar lavage fluid (BALF) was strikingly increased (mean 3.7, range 0.4 to 10 micrograms/ml). The value of C1q/albumin was several to 20 times greater in BALF than in serum samples from individual patients. In contrast, BALF samples from control subjects (ten patients with sarcoidosis and nine normal subjects) contained an undetectable amount (less than 0.02 micrograms/ml) of C1q. C3 in BALF also increased in the summer-type HP patients. Furthermore, C1q (as well as specific IgG and IgA antibody activities to Trichosporon cutaneum antigen) in BALF correlated with clinical symptoms and diffusing capacity (DCO), while the BAL lymphocytosis or the change of OKT4/OKT8 ratio did not. These findings are indicative of local secretion or concentration mechanism of C1q and C3, supporting the involvement of immune complexes in the respiratory tract of the patients.     

Trichosporon antibodies (corrected of antigen) and HLA-antigen in summer-type hypersensitivity pneumonitis in a family]

We encountered a family in which all of the three members (the parents, a 45-year-old woman and 51-year-old man, and their 15-year-old daughter) had Trichosporon cutaneum antibodies (corrected of antigen), and two (the parents) suffered from summer-type hypersensitivity pneumonitis in the late summer. The husband complained of dry cough, fever and dyspnea on exertion from July after severe interstitial pneumonitis and was treated with steroid pulse in September 2004. His wife visited our hospital and complained of a common cold-like symptom which progressed in August 2005. The couple were given diagnoses of summer-type hypersensitivity pneumonitis because they were positive for serum anti-Trichosporon mucoides antigen and asahii antigen. Their asymptomatic daughter was positive for these antigens. Both wife and daughter had HLA-DQ 8 (3) and 9 (3) that are suggested to be important HLA antigens related to the occurrence of summer-type hypersensitivity pneumonitis.     

A raised level of soluble CD8 in bronchoalveolar lavage fluid in summer-type hypersensitivity pneumonitis in Japan.

We used ELISA to measure soluble CD8 (sCD8) in the bronchoalveolar lavage fluid (BALF) and serum of patients with summer-type hypersensitivity pneumonitis (HP). The sCD8 levels in BALF were significantly higher in the patients with summer-type HP, surpassing those found in sarcoidosis and the other pulmonary diseases studied; however, the sCD8 levels in the serum of patients with summer-type HP did not differ from the levels of the healthy controls. The numbers of CD8+ T cells were increased in the BALF of the patients with summer-type HP, and there was a correlation between the sCD8 levels and the concentrations of CD8+ T cells. Gel filtration and polyacrylamide gel electrophoresis of the fluid revealed that the anti-CD8 monoclonal antibody-reactive components in the BALF of patients with pneumonitis corresponded to a protein with a molecular weight of between 52 and 54 kDa. Soluble CD8-rich fraction purified from the BALF of patients with summer-type HP augmented in vitro lymphocytes' proliferative responses stimulated with Cryptococcus neoformans, one of the causative agents for summer-type HP. Our result suggests that soluble CD8 in the BALF may play an important role in the pathogenesis of summer-type HP.     

Familial summer-type hypersensitivity pneumonitis in a grandfather and his two-and-a-half-year-old grandson]

We encountered a family in which two of the six members, the grandfather and his grandson, had summer-type hypersensitivity pneumonitis in late summer. Chest computed tomography of these patients showed small, diffuse nodular shadows in both lung fields, with mosaic-like opacities in the grandfather and ground glass opacity of both lung fields in the grandson. Bronchoalveolar lavage fluid from the grandfather disclosed high total cell counts, high percentages of lymphocytes, and a low ratio of CD 4+ cells to CD 8+ cells. Transbronchial lung biopsy specimens obtained from the same patient revealed alveolitis with non-caseous epithelioid cell granulomas in the interstitium and Masson bodies in the alveolar septa. These two patients recovered spontaneously after hospital admission. They had positive results in provocation tests for their home and were positive for serum anti-Trichosporon cutaneum (T. asahii, T. mucoides) antibodies. Both cases were accordingly diagnosed as summer-type hypersensitivity pneumonitis.     

Importance of serotype-related antigen in the induction of experimental hypersensitivity pneumonitis by Trichosporon cutaneum in rabbits. Correlation between granulomatous alveolitis and cellular and humoral immune responses to polysaccharide-rich antigen.

We evaluated the immunopathogenic properties of Trichosporon cutaneum, a major etiologic agent of Japanese summer-type hypersensitivity pneumonitis (HP). When the culture filtrate antigen of T. cutaneum was chromatographed on DEAE-cellulose, two peaks of polysaccharide, fractions A and B, were obtained. Fraction B was highly reactive to the specific IgG and IgA antibodies in the serum and bronchoalveolar lavage fluid samples obtained from sensitized rabbits, whereas fraction A was mainly reactive to the IgG antibodies. When the rabbits sensitized by intratracheal injection with the particulate antigen of T. cutaneum were challenged intratracheally by these antigenic fractions, fraction B induced granulomatous alveolitis, but fraction A induced alveolitis rather than granuloma. Lymphocytes of the mediastinal lymph nodes responded prominently to fraction B, but less to fraction A, as assessed by proliferative response. Each of the fractions B obtained from two strains of different serotypes (TIMM 1573, serotype I and TIMM 1318, serotype II) was reactive in enzyme-linked immunosorbent assay to the serum samples from the rabbits sensitized with homologous antigen, but far less to that from the rabbits sensitized with heterologous antigen, that is, the antigenic specificity of fraction B was related to the serotype of T. cutaneum. Fractionation by gel filtration on Sepharose CL-4B revealed that the molecular weight of the antigenic components in fraction B was larger than 1,000,000 and that the components consisted mainly of polysaccharide.(ABSTRACT TRUNCATED AT 250 WORDS)     

Japanese summer-type hypersensitivity pneumonitis. Geographic distribution, home environment, and clinical characteristics of 621 cases

A survey of hypersensitivity pneumonitis in Japan was carried out by nationwide questionnaire. A total of 835 cases of hypersensitivity pneumonitis was recognized during the last decade from 1980 to 1989. Of the 835 cases, 621 (74.4%) were summer-type hypersensitivity pneumonitis. The number of cases diagnosed doubled in the latter half of the 10-yr period. The residence of the patients was mostly in the western and southern parts of the country; the northernmost incidence was in Akita Prefecture at a latitude of 40 degrees north. Of the patients 86% experienced initial symptoms from June to September with a peak in July. Female patients were 2.0 times as numerous as male patients, probably as a result of more exposure to the offending antigen at home since 39.8% of the patients were female homemakers without an outside occupation. The frequency of family occurrence was 23.8%. The mean age of the patients' houses was 20.5 yr after building, and more than half had unsanitary conditions, such as shady, damp, or poorly ventilated rooms. Serum anti-Trichosporon cutaneum antibody activities were positive in 260 (99.2%) of the 262 cases examined. We conclude that summer-type hypersensitivity pneumonitis occupies the majority of hypersensitivity pneumonitis in the country and that to prevent the disease concentrated attention should be given to the home environment from the viewpoint of environmental medicine.     

A familial case of summer-type hypersensitivity pneumonitis possibly associated with bird breeder's lung diagnosed by bronchoalveolar lavage fluid]

Case 1: A 32-year-old woman had cough and exertional dyspnea in August 2002, and chest computed tomographic scan revealed diffuse centrilobular nodules. Bronchoalveolar lavage fluid (BALF) showed a high proportion of lymphocytes with a decreased CD 4/CD 8 ratio. Transbronchial lung biopsy (TBLB) specimens showed alveolitis. Summer-type hypersensitivity pneumonitis was diagnosed on the basis of positive findings of anti-Trichosporon antibodies in the serum. Case 2: A 64-year-old man, the father of Case 1, also had cough and exertional dyspnea in August 2003. He had been in close contact with pigeons. Chest computed tomographic scan revealed bilateral map-like ground-glass opacities predominantly in the upper lobes. BALF showed a high proportion of lymphocytes with a decreased CD 4/CD 8 ratio. TBLB specimens showed alveolitis, granuloma and Masson body in the air spaces. Specific IgG and IgA antibodies against Trichosporon asahii, IgA antibodies against Trichosporon mucoides, and IgA antibodies against pigeon dropping extracts were found only in the BALF but not in the serum. Although a positive finding of returning-home provocation test was definitive in diagnosing summer-type hypersensitivity pneumonitis, he was also suspected of having bird fancier's lung.     

Hypersensitivity pneumonitis induced by Trichosporon cutaneum

We isolated Trichosporon cutaneum from the houses of patients with hypersensitivity pneumonitis and investigated its possible role as an etiologic agent in 21 cases of the disease. A high titer of anti-T. cutaneum antibody was observed in serum samples from 20 (95.2%) of the 21 patients, but only a low titer was found in less than 9% of the control serum samples. Anti-Cryptococcus neoformans antibody was also observed in the patients' serum samples, but it appeared, in most samples, to be a cross-reaction to T. cutaneum antigen. This was because the anti-C. neoformans antibody was completely absorbed with T. cutaneum, but the anti-T. cutaneum antibody was only partly absorbed with C. neoformans. Besides, no C. neoformans was isolated from the patients' environment. A bronchoprovocation test with T. cutaneum was done on one patient, with a positive result. We also found T. cutaneum in bird droppings containing nutrients for this organism. To our knowledge, this is the first report on hypersensitivity pneumonitis induced by T. cutaneum.     

IgA and IgG antibody activities of serum and bronchoalveolar fluid from symptomatic and asymptomatic pigeon breeders

Serum IgA and IgG antibody activities against pigeon serum were measured in 16 symptomatic pigeon breeders, 20 asymptomatic pigeon breeders, and 3 normal subjects by radioimmunoassay. The IgA and IgG antibody activities against pigeon antigen of the group of patients with disease was significantly greater than those of patients in the asymptomatic and the control group. The overlap of results for the symptomatic and asymptomatic breeders limits the diagnostic value of these individual IgA or IgG antibody determinations. Bronchoalveolar fluid and serum samples from a smaller group of pigeon breeders who underwent lung lavage were available for studies of antibody activity against pigeon serum. Ten asymptomatic and 6 symptomatic breeders were available for study. Both IgG and IgA antibody activities were detected by radioimmunoassay in serum samples and bronchoalveolar fluid. The IgA antibody activity determined by the radioimmunoassay was higher in the respiratory secretions.     

A case of severe summer-type hypersensitivity pneumonitis treated with high-dose administration of steroid]

A 40-year-old man who lived in a wooden house built 30 years ago presented with complaints of fever, dry cough and dyspnea. Chest X-ray findings showed interstitial shadows throughout bilateral lung fields. After admission, high-dose administration of 3000 mg of methylprednisolone was performed because of deterioration of chest X-ray shadows and symptoms. In a week, clinical data and symptoms improved. Findings of BAL fluid on admission revealed a relative increase of lymphocytes, neutrophils and mast cells, and pathological findings of transbronchial lung biopsy revealed non-caseous granulation and alveolitis. Precipitating antibodies and indirect fluorescent antibodies against Trichosporon cutaneum and Cryptococcus neoformans had positive reactions and T. cutaneum was isolated and identified from the patient's house. A diagnosis of summer-type hypersensitivity pneumonitis was made according to the criteria advocated by Ando et al. This seemed to be a rare case of summer-type hypersensitivity pneumonitis prolonged after isolation from his normal living environment, successfully treated by high-dose administration of steroid.     

A case of summer-type hypersensitivity pneumonitis with bronchoalveolar lavage performed 4 years before onset

A 51-year-old man with chief complaints of cough, fever, and dyspnea was admitted to our hospital. Based on a home provocation test, transbronchial lung biopsy specimens, and a serum antibody, we diagnosed summer-type hypersensitivity pneumonitis. In 1983 when the patient was 46 years old, thymectomy was performed for thymoma. Prior to surgery, bronchoalveolar lavage (BAL) was performed. Total cell count and neutrophils had already increased in BALF. Furthermore, the increase in BALF cell neutrophil count was also seen at the time of admission and after the home provocation test. Because an increase of neutrophils in BALF cells was seen not only at onset but before onset, further studies are required to clarify the role of neutrophils and the factors that increase them in hypersensitivity pneumonitis.     

A case of acute hypersensitivity pneumonitis caused by contact with budgerigars (bird-breeder's lung)]

A 58-year-old woman was admitted to our hospital for repeated episodes of dry cough, low-grade fever, and gradual development of dyspnea on exertion. Chest computed tomography showed diffuse ground-glass opacities in both lung fields. Bronchoalveolar lavage fluid (BALF) showed an increased number of lymphocytes, and transbronchial lung biopsy revealed alveolitis and epithelioid cell granuloma. The acute onset and the patient's living environment suggested summer-type hypersensitivity pneumonitis. However, anti-Trichosporon antibody was negative and a definitive diagnosis could not be made. The patient's condition improved with corticosteroid treatment and, after discharge from the hospital, she moved to a new home. The following year, however, her symptoms returned, and she was readmitted to our hospital. Recurrence of the disease despite the change in environment ruled out summer-type hypersensitivity pneumonitis. Given the patient's history of budgerigar breeding five years earlier, bird breeder's lung was suspected. Anti-bird excreta antibody found in the patient's serum and BALF, along with a positive lymphocyte stimulation test against pigeon serum, strongly suggested an acute onset of bird breeder's lung. We report a rare case of acute bird breeder's lung with radiologic findings of ground-glass opacities, a one-year disease-free period and an acute relapse.     

Familial summer-type hypersensitivity pneumonitis--case report and review of literature]

A 37-year-old-woman was admitted to our hospital because of chest bilateral reticular shadow with fever, cough, general malaise and exertional dyspnea in the summer. A diagnosis of summer-type hypersensitivity pneumonitis (SHP) was made by radiological, serological and histological examinations. Her 10-year-old daughter had chest reticular shadows and similar symptoms. These two patients were positive for serum anti-Trichosporon cutaneum (T. asahii, T. mucoides) antibodies and T. asahii was identified from cultured samples obtained from their house. They recovered spontaneously after hospitalization or isolation from the antigen. We reviewed the clinical features in sixteen families with familial SHP reported in Japan. Children aged under 15 years old accounted for 34% and there was no gender difference among patients. This finding differs from the conventionally defined features of patients with SHP. Measurements of serum KL-6, SP-D and SP-A seem to be useful for auxiliary diagnosis and monitoring the disease activity of SHP, especially in pediatric cases who cannot undergo invasive evaluation.     

A case of hypersensitivity pneumonitis caused by Gyrodontium versicolor]

A 36-year-old woman was admitted to our hospital because of fever, dry cough, dyspnea on exertion and body weight loss in August 2000. Chest radiography and CT scanning showed diffuse ground glass opacity and small centrilobular nodules in the middle and lower lung fields of both lungs. Serum antibody against Trichosporon cutaneum was positive; and summer-type hypersensitivity pneumonitis was therefore initially diagnosed. Treatment with methylprednisolone and prednisolone decreased the symptoms, but the dyspnea reappeared when the patient was at home. Inspection of her house revealed the presence of fungi under the floor. After these were removed, her symptoms disappeared completely. The lymphocytic stimulation test of the peripheral blood was positive for the fungi, and it was therefore suggested that they were the cause of her hypersensitivity pneumonitis. The fungi were identified as Gyrodontium versicolor. This is the first report of hypersensitivity pneumonitis caused by Gyrodontium versicolor.     

TA-19, a novel protein antigen of Trichosporon asahii,in summer-type hypersensitivity pneumonitis

The most common form of hypersensitivity pneumonitis in Japan is summer-type hypersensitivity pneumonitis (SHP), which is caused by the inhalation of Trichosporon asahii or Trichosporon mucoides. To seek protein antigens relevant to the immunopathogenesis of SHP, we constructed a cDNA expression library of T. asahii, a major causative yeast species of SHP. Using the immunoscreening method, we identified and cloned a novel gene encoding a 19-kD protein, named TA-19, which proved to be specifically recognized in the bronchoalveolar lavage (BAL) fluids and sera of patients with SHP. IgG, IgA, and IgM antibodies to the recombinant TA-19 protein were significantly elevated in the sera as well as in the BAL fluids from SHP patients compared with those from non-SHP groups. This protein also induced SHP-specific proliferation of the mononuclear cells from both the peripheral blood and BAL. These results reveal that TA-19 derived from T. asahii may play a relevant role in specific cellular and humoral immune responses in patients with SHP.     

Pigeon 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-labile, 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.     

A case of Japanese summer-type hypersensitivity pneumonitis: monitoring with serum KL-6 and examination of the phenotype of HLA]

A 55-year-old woman was admitted with a cough and fever in August. A diagnosis of Japanese summer-type hypersensitivity pneumonitis was made on the basis of radiological, serological and pathological findings, in addition to positive returning home provocation. Serum KL-6 was monitored during the clinical course. Although KL-6 fluctuated slowly in comparison with the clinical symptoms and HRCT findings, it was considered useful for confirming the effects of treatment. Serum anti-Trichosporon antibody and the phenotype of HLA were studied in both the patient and her asymptomatic roommate, with whom she had no blood relationship. Though both were sensitized immunologically, HLA-DQ 3, which was reported to be associated with Japanese summer-type hypersensitivity pneumonitis, was detected in the patient but not in her roommate. It was suggested that HLA plays a role in the development of this disease.     

Two cases of budgerigar breeder's lung

We report 2 patients with Bird breeder's lung associated with keeping budgerigars. Patient 1 kept a budgerigar indoors for 13 years. Histological examination revealed fibrosis and granulomatous changes in the alveolar septa. Among T lymphocyte subsets in BALF, helper T cell were increased. In the serum, precipitating antibodies to budgerigar dropping extract were observed. Patient 2 had kept Budgerigar for 7 months. Histologically, alveolitis was the primary finding, and suppressor T cell in the BALF were increased. Precipitating antibodies to budgerigar dropping extract was observed only in the BALF but not in serum. In patient 1 who showed a chronic course and granuloma formation, helper T cell in the BALF were increased while in patient 2 who showed an acute course and alveolitis, suppressor T cell in the BALF were increased, and precipitating antibody was observed only in the BALF. These findings suggest that T lymphocyte subsets in the BALF differ depending on the stage of hypersensitivity pneumonitis, and specific local antibody production precedes systemic antibody production.     

A study on interleukin 2 and soluble interleukin 2 receptor in serum and bronchoalveolar lavage fluid from patients with farmer's lung]

Interleukin 2 (IL-2) and soluble Interleukin 2 receptor (IL-2R) were assayed in serum and bronchoalveolar lavage fluid (BALF) from patients with farmer's lung. Patients with farmer's lung had significantly higher levels of soluble IL-2R in serum and BALF than normal controls. The level of soluble IL-2R was correlated with the disease activity, and fell to within the normal range after successful treatment with corticosteroids. On the other hand, asymptomatic dairy farmers with precipitating antibodies against Micropolyspora faeni antigen or Thermoactinomyces vulgaris antigen had the same levels of soluble IL-2R in their serum and BALF as normal controls. IL-2 levels in serum and BALF were significantly higher in patients with farmer's lung than in normal controls; however, asymptomatic patients had low levels comparable to those of normal controls. These results indicate that soluble IL-2R could be a sensitive parameter for evaluation of the activity of this disease. It is also suggested that the IL-2/IL-2R system is important in the pathogenesis of hypersensitivity pneumonitis.