Localization of inhaled trimellitic anhydride to lung with a respiratory lymph node antibody secreting cell response.

Male Sprague-Dawley rats were exposed to trimellitic anhydride (TMA) by inhalation (500 micrograms/m3), 4 hours a day, for 1 to 10 days. TMA was localized to lung cells by immunoelectron microscopy. Enzyme-linked immunosorbent assay (ELISA) and Western blot analysis were used in an attempt to localize TMA to lung lavage proteins. The lung-associated lymph node (LALN) B-lymphocyte response was measured by quantitation of immunoglobulin (Ig)G, IgA, and IgM antibody secreting cells specific for TMA rat serum albumin (TM-RSA) by use of the enzyme-linked immunospot assay (ELISPOT) method. The IgG, IgM, and IgA antibody response to TM-RSA in serum and lavage fluid was quantitated by ELISA. Lung injury was assessed by the number of external lung hemorrhagic foci and lung weight. Immunoelectron microscopy localized TMA to alveolar and bronchial cells on all exposure days. ELISA detected trace amounts of TMA haptenized lavage proteins that could not be detected by Western blot analysis. A marked increase occurred in lung injury from day 7 to 10. The LALN IgG, IgA, and IgM antibody secreting cell response to TM-RSA paralleled measures of lung injury. IgG, IgM, and IgA serum and lavage antibody to TM-RSA were correlated with lung injury measures. Lavage and serum IgG antibody levels had the highest correlation with lung injury.

A clinical and immunologic study of employees in a facility manufacturing trimellitic anhydride.

We conducted a 1-year cross-sectional survey of 474 employees of a large chemical manufacturing complex to relate trimellitic anhydride (TMA) exposure to serologic and clinical outcomes. In 1988-1989, employees were evaluated by history and immunologic assay of total (T) and IgE antibody to trimellityl human serum albumin (TM-HSA). All employees were assigned to a TMA exposure class, from 1 (highest) to 5 (lowest), by an industrial hygienist, independent of the clinical and immunological data. Thirty-two (6.8%) of 474 employees had a TMA immunologic syndrome, 31.6% with an irritant response and 61.6% with no symptoms. Twelve had asthma/rhinitis, 10 had the late respiratory systemic syndrome (LRSS), four had late onset asthma, one had late onset arthralgia, and five had a distant history of LRSS. Included in this survey of the entire work force were 321 new enrollees, who had not joined the previous (1976 to 1988) voluntary surveillance program. Only four (1.3%) of the new enrollee group had a TMA immunologic syndrome. Among new enrollees, there were lower mean total and IgE serum antibody levels in lower exposure classes and a higher percentage with elevated antibody levels in high exposure classes (for T, x2 = 17.5, p = .0016; for IgE, x2 = 76.7, p less than .0001). In the new enrollee population, demographic variables of age, sex, date of hire, and smoking status were examined related to antibody levels. Only current or former smoking was related to elevated total antibody levels.

A twelve-year clinical and immunologic evaluation of workers involved in the manufacture of trimellitic anhydride (TMA).

The spectrum of immunologic lung disease occurring in a population of 196 workers involved in the manufacture of trimellitic anhydride (TMA) was assessed from January 1976 through December 1987. Workers were evaluated clinically by history, blood counts and chemistries, chest x-ray, and pulmonary function studies. Immunologic tests included skin testing with trimellityl-human serum albumin (TM-HSA) and assay of total antibody (TA) and of IgE antibody binding of 125I-TM-HSA. Seventeen workers had IgE-mediated asthma/rhinitis with a positive prick test to TM-HSA and IgE antibody of 0.8-57 ng of TM-HSA bound/ml. Seven workers had a late respiratory systemic syndrome with TA from 760-56,000 ng of TM-HSA bound/ml. Four had both syndromes. Three had late onset asthma with TA of 3,700-10,000 and trace levels of IgE to TM-HSA. One had marked arthralgia and myalgia occurring hours after TMA exposure, without respiratory symptoms, with an elevated TA level of 24,000. Of 46 workers reporting no symptoms, 8% had low TA levels, while 16% of 113 with irritant symptoms had low TA levels. There was a reduction in the number of workers exhibiting an immunologic syndrome during 1982-1987-8% (7 of 85) compared to 23% (26 of 111) during 1976-1981--in spite of increased TMA production. This paralleled environmental control and worker education efforts. Cooperative research by an academic Allergy-Immunology program and industry has defined TMA clinical syndromes and provided methods of immunologic monitoring, resulting in a reduction in new cases in spite of increased production of TMA.

Lung injury induced by short-term intermittent trimellitic anhydride (TMA) inhalation.

We have developed a rat model of lung injury with interstitial pneumonitis, lung hemorrhage, and a systemic and pulmonary immune response to trimellitic anhydride (TMA)-haptenized proteins induced by TMA inhalation for 10 days. The present studies explored the induction of lung injury induced by short-term intermittent TMA inhalation, a model more likely to simulate short-term industrial exposures during inadvertent spills of TMA. Sprague-Dawley rats inhaled TMA powder (500 micrograms/m3) on days 1, 5, and 10, and were necropsied on day 30, 18 hours after a 6-hour TMA-inhalation challenge on day 29. Rats were bled every second day and at necropsy. Serum IgG, IgA, and IgM antibody to trimellityl rat serum albumin was measured by ELISA. There was a rise in IgM and IgA antibody to trimellityl rat serum albumin starting at day 5 that peaked at day 20 with a decline in IgM by day 30. IgG antibody rose at day 7, peaked at day 20, and plateaued. The IgG antibody level was 10 times higher than the IgA or IgM level. In a second experiment, 18 rats were administered TMA-inhalation exposure on days 1, 5, and 10, and a TMA challenge on day 22. The number of hemorrhagic foci, lung weights, and lung-displacement volumes at necropsy on day 23 were highly correlated with IgG, IgA, and IgM serum-antibody levels. In a final experiment, rats developed a mean of 112 hemorrhagic foci per lung on day 30 after receiving only two TMA-inhalation exposures on days 1 and 5 with a rechallenge on day 29. (ABSTRACT TRUNCATED AT 250 WORDS)

Antibody response to trimellityl hemoglobin in trimellitic anhydride-induced lung injury.

Sprague-Dawley rats were exposed to trimellitic anhydride by inhalation, and the antibody response to trimellityl (TM)-conjugated hemoglobin (HB) and TM rat serum albumin (RSA) was compared. Groups of rats were exposed to trimellitic anhydride by inhalation 6 hours per day for 2, 6, or 10 days at 100 micrograms/m3 and compared to a control group exposed to filtered air. The IgG antibody response to TM-HB in both serum and bronchoalveolar lavage (BAL) fluid was measured with ELISA. IgG antibody levels to TM-HB rose significantly throughout the exposure. A positive correlation was found between IgG to TM-HB in serum and BAL fluid. In addition, this response in both serum and BAL fluid correlated with the IgG antibody response to TM-RSA. Cross-inhibition studies indicated the existence of shared antigenic determinants on TM-RSA and TM-HB. The IgG antibody to both antigens was specific for new antigenic determinants and not for the TM hapten.

A serial immunologic and histopathologic study of lung injury induced by trimellitic anhydride.

Trimellitic anhydride (TMA) can induce immunologic lung disease in exposed workers. We have developed a rat model of TMA lung injury characterized by lung hemorrhage and an immune response to trimellityl (TM) haptenized lung proteins. The model is similar to the pulmonary disease-anemia syndrome (PDA) seen in workers exposed to TMA fumes. Sprague-Dawley rats, 15 per exposure period, inhaled micronized TMA powder, 100 micrograms/m3, 6 h/day, for 2,6, or 10 days and were sacrificed. At each time period, total, IgG, IgA, and IgM antibody to TM-rat serum albumin (TM-RSA) were measured by radiolabeled antigen binding and enzyme-linked immunosorbent assay (ELISA) in serum and bronchoalveolar lavage fluid (BAL). Hemorrhagic lung foci, weight, and displacement volume were determined, and lungs were examined by light and electron microscopy. There was no lung injury or antibody response at 2 days. There was minimal lung injury at 6 days with low levels of antibody in BAL and serum. At 10 days, there was a marked increase in hemorrhagic foci and in BAL and serum antibody levels. BAL antibody levels at 6 and 10 days had higher correlations with measures of lung injury than corresponding serum levels. There was minimal ultrastructural change at 6 days. By Day 10, there was marked intraalveolar hemorrhage, alveolar septal inflammatory nodules, abundant alveolar macrophages, and evidence of endothelial and epithelial cell injury. These results indicate that the immune response to inhaled TMA occurs parallel with the development of lung lesions, and antibody levels in BAL and serum are highly correlated with lung injury.

Levels and specificity of antibody in bronchoalveolar lavage (BAL) and serum in an animal model of trimellitic anhydride-induced lung injury.

A study was undertaken to characterize the antibody response in rats exposed to trimellitic anhydride (TMA) by inhalation. Total antibody levels directed to trimellitic rat serum albumin (TM-RSA) from TMA-exposed rats were assayed by an ammonium sulfate technique. Total antibody levels in bronchoalveolar lavage (BAL) and the matched serum were compared by correction for the albumin content of each. An ELISA was developed to detect IgG, IgA, and IgM directed toward TM-RSA in BAL and serum and to compare class-specific antibody levels in BAL and serum by normalizing for albumin content. The specificity of the rat IgG response was determined by ELISA inhibition with TM-RSA and TM-human serum albumin (TM-HSA) and compared with reciprocal inhibition studies with serum from TMA-exposed workers. The levels of total antibody in BAL were three to 15 times greater than the levels found in the matched serum pair. IgG, IgA, and IgM antibodies were detected in the BAL and the serum of TMA-exposed rats but not in control rats. In each of the four rats tested, all antibody classes were present in equal or greater amounts in the BAL than in the serum. Complete inhibition of the rat IgG binding in ELISA was observed when TM-RSA or TM-HSA were added as inhibitors. Human IgG was inhibited in ELISA only by TM-HSA. In an animal model of human lung disease, the levels of total antibody as well as class-specific antibodies directed against TM-RSA were greater in BAL than in serum.(ABSTRACT TRUNCATED AT 250 WORDS)

A model of immunologic lung injury induced by trimellitic anhydride inhalation: antibody response.

We studied lung injury induced in Sprague-Dawley rats by trimellitic anhydride (TMA) inhalation. Groups of 40 male and 20 female rats were exposed to TMA by inhalation at target concentrations of 0, 10, 30, 100, and 300 micrograms/m3, 6 hours per day, 5 days per week, for 2 weeks. Rats in each exposure group were sacrificed after 10 exposures or rested for 12 days and either sacrificed or received a 6-hour TMA challenge at their initial exposure levels and sacrificed at 24 hours. At each sacrifice, serum antibody to radiolabeled trimellityl rat serum albumin (RSA-TM) was measured by an ammonium sulfate technique, and lung pathology was determined. After 10 days of exposure, external hemorrhagic lung foci were directly related to the exposure concentration of TMA. Serum antibody binding of RSA-TM correlated with exposure concentration, hemorrhagic lung foci, and lung weight. There was healing of lung lesions 12 days after exposure with a return of lung lesions only 18 hours after the 6-hour inhalation challenge. A correlation between serum antibody to RSA-TM, hemorrhagic foci, and lung weight existed after challenge. This model clarifies two clinical entities observed in exposed workers, the late respiratory systemic and the pulmonary disease-anemia syndromes.

Specific bronchoalveolar lavage IgG antibody in hypersensitivity pneumonitis from diphenylmethane diisocyanate.

We evaluated a patient for dyspnea, fever, malaise, and hypoxemia that developed after exposure to diphenylmethane diisocyanate (MDI). Specific inhalation challenge with MDI caused fever, leukocytosis, a restrictive decline in forced vital capacity, and a decrease in Pao2 several hours after challenge. Bronchoalveolar lavage 24 h after challenge showed lymphocytic alveolitis. Specific IgG antibodies to MDI human serum albumin (MDI-HSA) conjugate were demonstrated in serum and bronchial lavage fluid using the enzyme-linked immunoabsorbent (ELISA) technique. These findings suggest participation of both humoral and cellular immunity in the pathogenesis of hypersensitivity pneumonitis from isocyanate exposure.

IgG antibody to purified protein derivative by enzyme-linked immunosorbent assay in the diagnosis of pulmonary tuberculosis.

We studied the diagnostic utility of an enzyme-linked immunosorbent assay (ELISA) in hospitalized patients with suspected pulmonary tuberculosis (TB). A positive culture for M. tuberculosis identified active disease, and 3 negative cultures and smears defined the negative group. IgG antibody activity was determined by adding a 1:1,000 dilution of serum to plates coated with PPD antigen. Alkaline phosphatase labeled anti-IgG was added, color developed, and an optical density index (ODI) was determined. Twenty-one patients with M. tuberculosis TB had a mean ODI of 0.27, which was higher than 99 patients without TB, ODI 0.10 (p less than 0.001). An ODI of 0.15 or greater was established as a positive ELISA test. For patients with M. tuberculosis TB, the ELISA had a sensitivity of 67 and a specificity of 79%. The first smear had a sensitivity of 57 and a specificity of 99%. With the first smear and the ELISA test results, a combined sensitivity of 86% was achieved. When both ELISA and the first smear were negative (101 cases), active TB caused by M. tuberculosis was found in only three patients (3.0%).

The relationship of total serum IgE and blocking antibody in trimellitic anhydride--induced occupational asthma.

Clinical and immunologic evaluations were performed on workers who manufacture trimellitic anhydride (TMA), a chemical used widely in the plastics industry. Serum samples of 29 workers exposed to TMA dust and fumes were assayed for total IgE, as well as specific IgE and total antibody binding to radiolabeled trimellityl human serum albumin (TM-HSA). Fifteen workers were determined to have significant specific IgE to TM-HSA (greater than or equal to 1 ng of 125I-TM-HSA bound per milliliter). Of these 15 individuals, seven workers had immediate asthma and rhinitis and positive cutaneous prick tests and eight were nonasthmatic and skin test negative. Mean peak total serum IgE was significantly higher in the prick test positive group (p less than 0.02) and higher than in 12 exposed asymptomatic IgE antibody-negative workers (p less than 0.001). Total serum IgE followed rises and falls in TM-HSA--Specific IgE binding, correlating to the onset of allergic symptoms in all seven workers. Total antibody binding of 125I-TM-HSA, as determined by the ammonium sulfate precipitation technique, was higher in the skin test-negative group of eight (mean 12,062 ng per milliliter 125I-TM-HSA bound) than in the seven symptomatic skin test-positive workers (mean 2930, p less than 0.004). Determinations of total antibody divided by specific IgE (both in nanograms of 125I-TM-HSA bound per milliliter) or blocking ratios clearly discriminated the seven skin test-positive workers with TMA-induced asthma (range 10 to 666) from the eight skin test-negative workers (range 1580 to 16,000).(ABSTRACT TRUNCATED AT 250 WORDS)

The enzyme-linked immunosorbent assay method for IgG antibody to purified protein derivative in cerebrospinal fluid of patients with tuberculous meningitis.

Three patients with culture-proven Mycobacterium tuberculosis meningitis were studied. Analysis of cerebrospinal fluid with an enzyme-linked immunosorbent assay (ELISA) method measuring IgG antibody to purified protein derivative rapidly yielded positive results, whereas results of acid-fast smears were negative and cultures took several weeks before growth appeared. We did serial studies of cerebrospinal fluid and sera from one patient. Initially, greater amounts of IgG antibody to purified protein derivative were present in the cerebrospinal fluid than in the serum. The antibody level in the cerebrospinal fluid paralleled the patient's clinical course, cerebrospinal fluid cell count, protein level, and glucose level. Cerebrospinal fluid samples from 33 hospitalized control patients were negative for antibody to purified protein derivative. The ELISA method measuring IgG antibody to purified protein derivative should be evaluated as a means of early diagnosis and management of patients with suspected tuberculous meningitis.

Combined alveolitis and asthma due to hexamethylene diisocyanate (HDI), with demonstration of crossed respiratory and immunologic reactivities to diphenylmethane diisocyanate (MDI).

A worker exposed intermittently to hexamethylene diisocyanate (HDI) developed episodes of dyspnea, wheezing, and fever on working days. Complete lung function tests performed when the subject was asymptomatic were normal except for increased airway responsiveness to histamine, which significantly improved after a 3 wk period off work. At that time, specific inhalation challenges with HDI were carried out. After being exposed for 5 min, the subject developed general malaise, cough, fever, and leukocytosis, together with a mixed restrictive and obstructive breathing defect. We demonstrated a subsequent increase in airway hyperexcitability, which lasted for 2 mo. The subject was also challenged with diphenylmethane diisocyanate (MDI) for 15 min. A late obstructive reaction was documented. Increased levels of specific IgG antibodies against HDI-human serum albumin (HSA) and MDI-HSA were demonstrated.

Use of an enzyme-linked immunosorbent assay technique in the differential diagnosis of active pulmonary tuberculosis in humans.

Sera from patients with active pulmonary tuberculosis and pulmonary diseases frequently mimicking tuberculosis were assayed for immunoglobulin G antibody activity to purified protein derivative (PPD) by an enzyme-linked immunosorbent assay. A method of standardization was developed to limit assay variation. Patients with active pulmonary tuberculosis had a significantly greater mean level of antibody than had patients with atypical tuberculosis (P = 0.005), sarcoidosis (P = 0.0001), histoplasmosis (P = 0.004), blastomycosis (P = 0.008), or cryptococcosis (P = 0.017), patients who had received bacille Calmette-Guérin vaccination (P = 0.003) or who had a history of treated tuberculosis (P = 0.003), and PPD skin test-positive and skin test-negative control subjects (P = 0.001). This technique may have potential use as a rapid diagnostic aid in evaluating patients with suspected active pulmonary tuberculosis.

Syndromes in workers exposed to trimellitic anhydride. A longitudinal clinical and immunologic study.

A longitudinal study of workers involved in the manufacture of trimellitic anhydride has been in progress since 1977. Trimellitic anhydride is a low-molecular-weight, reactive chemical used in the manufacture of plastics. Initial studies done in 1976 defined three syndromes due to inhalation of trimellitic anhydride: asthma-rhinitis, a late respiratory systemic syndrome, and an irritant response. Also, serologic techniques were developed to measure total antibody and IgE antibody to trimellityl human serum albumin. From 1977 to 1981, 64 workers were assessed. Six workers presented with asthma-rhinitis, high levels of IgE antibody, and skin test reactivity to trimellityl human serum albumin; five workers developed the late respiratory systemic syndrome; and one worker had both immunologic syndromes. These serologic measurements detected or were predictive of an immunologic respiratory illness and were useful in monitoring workers involved in the manufacture of trimellitic anhydride.

Detection of immunoglobulin G antibody to purified protein derivative in patients with tuberculosis by radioimmunoassay and enzyme-linked immunosorbent assay.

Sera from patients with active tuberculosis and sera from appropriate control individuals were assayed for immunoglobulin G antibody activity to purified protein derivative by a polystyrene tube radioimmunoassay and an enzyme-linked immunosorbent assay. Both assays showed a marked increase in immunoglobulin G antibody activity in patients with active tuberculosis. There was no overlap between the values for the patient group and the values for the purified protein derivative skin test-positive control individuals. The replication of these assays was excellent, and both could provide quantitative measurements of immunoglobulin G antibody activity to purified protein derivative antigen within 24 h. These techniques have potential as rapid diagnostic aids in evaluating patients with suspected active tuberculosis.

Occupational asthma secondary to inhalation of garlic dust.

An atopic patient in whom the inhalation of occupationally encountered garlic dust precipitated asthma is reported. Studies revealed the presence of immediate skin-test reactivity to garlic extract, and specific IgE anti-garlic antibodies were detected in the patient's serum by the polystyrene tube solid phase radioimmunoassay (PTRIA) technique. Bronchial challenge with garlic dust also resulted in an immediate asthmatic response typical for an IgE-induced mechanism.

Comparison of the radioallergosorbent test and a quantitative solid-phase radioimmunoassay for the detection of ragweed-specific immunoglobulin E antibody in patients undergoing immunotherapy.

Commercially available radioallergosorbent test (RAST) was compared with a quantitative solid-phase polystyrene tube radioimmunoassay (PTRIA) for the detection of ragweed-specific IgE antibody in sera of patients undergoing ragweed immunotherapy. Both assays showed good replication over a wide range of IgE antibody activity. The RAST was limited in the ability to discriminate between sera with high levels of specific IgE antibody. In addition, RAST gave falsely low values of IgE antibody activity in the presence of high levels of blocking antibody.

Seasonal variation of IgE antibody specific for ragweed antigen E (IgE-a-AgE) from the basophil surface in patients with ragweed pollenosis.

Seasonal elevation in total serum IgE and IgE antibody specific for antigen E (IgE-a-AgE) are well recognized in patients with ragweed pollenosis. We found that the ratio of specific IgE-a-AgE to total IgE in plasma was the only measure predictive of the number of specific IgE-a-AgE molecules per basophil. Although both total and specific IgE rose with season, the ratio did not change, the number of IgE-a-AgE molecules per basophil did not change, and histamine release in response to AgE did not increase.

Immunoglobulin E-mediated asthma and hypersensitivity pneumonitis with precipitating anti-hapten antibodies due to diphenylmethane diisocyanate (MDI) exposure.

Two workers are presented who were exposed to diphenylmethane diisocyanate (MDI) while coating pipes with a polyurethane foam. After a latent period of exposure, worker A developed immediate-onset asthma and worker B developed a clinical picture of hypersensitivity pneumonitis for which he was hospitalized. The antibody response of these workers to a conjugate of MDI with human serum albumin (MDI-HSA) was measured by gel precipitation, total antibody binding of 125I MDI-HSA, and specific IgG and IgE antibody by polystyrene-tube radioimmunoassay (PTRAI). Worker B had precipitating antibody to MDI-HSA by double immunodiffusion in gel. Both workers had high levels of IgG antibody specific for MDI-HSA which had some cross-reactivity with a conjugate or toluene diisocyanate and HSA. Total serum antibody binding of 125I MDI-HSA was 15 microgram/ml in worker A and 900 microgram/ml in worker B. Both workers had serum IgE antibody specific for MDI-HSA as measured by two PTRIA techniques. These results indicate that a marked immunologic response to MDI is possible in exposed workers and that hypersensitivity pneumonitis can occur subsequent to the inhalation of a low-molecular-weight chemical in the industrail setting.