Polystyrene Tube Radioimmunoassay Research Articles

Antibodies against toluene diisocyanate protein conjugates three methods of measurement

With the use of canine antisera against toluene diisocyanate (TDI)-dog serum albumin (DSA), techniques for measuring antibody against TDI-DSA were evaluated. The use of an ammonium sulfate precipitation assay showed suggestive evidence of antibody binding but high levels of TDI-DSA precipitation in the absence of antibody limit any usefulness of this technique. Double-antibody co-precipitation techniques will measure total antibody or Ig class antibody against 125I—TDI-DSA. These techniques are quantitative. The polystyrene tube radioimmunoassay is a highly sensitive method of detecting and quantitatively estimating IgG antibody. The enzyme linked immunosorbent assay is a rapidly adaptable method for the quantitative estimation of IgG, IgA, and IgM against TDI-homologous proteins. All these techniques were compared and results are demonstrated by using the same serum sample for analysis.

Intractable sneezing due to IgE-mediated triethanolamine sensitivity

Sneezing has been commonly recognized as part of the nasal allergic reaction in response to pollen and inhalant allergens; isolated intractable sneezing is an unusual presentation in allergic patients. An 8-yr-old white girl developed intractable sneezing in the fall while walking past an area where fresh roofing tar was being prepared. Her personal and family histories were negative for allergic disease or symptoms. Positive physical findings were boggy nasal mucosa; pollen and inhalant skin tests were negative. Response to topical cromolyn and beclomethasone administered intranasally was only partial; antihistamines were little help. Upon careful review of history, exposure to clothes washed in Miracle White Laundry Soil and Stain Remover correlated to symptoms; removal from clothing by extensive washing relieved the sneezing, which recurred upon exposure. Prick test was positive to triethanolamine at 10 − 7 M to 10 − 4 M and not to other ingredients of this product. Investigation demonstrated dose-dependent leukocyte histamine release (25% to 27% specific release) to triethanolamine (10 − 4 M to 10 − 7 M); this release (50% at 10 − 5 M) was inhibited by preincubation with cromolyn sodium (5 × 10 − 6 M). Passive cutaneous anaphylaxis was demonstrated to triethanolamine (10 − 7 M to 10 − 4 M); specific IgE to triethanolamine was demonstrated by polystyrene tube radioimmunoassay. Controls had no histamine release or specific IgE. Thus exposure to triethanolamine caused IgE-mediated intractable sneezing.

Clinical and immunologic evaluation of trimellitic anhydride- and phthalic anhydride-exposed workers using a questionnaire with comparative analysis of enzyme-linked immunosorbent and radioimmunoassay studies

Twenty workers exposed to trimellitic anhydride (TMA) powder an phthalic anhydride (PA) fumes concurrently were evaluated by questionnaire and serum antibody studies. The study group was divided into high-an low-exposure groups on the basis of job descriptions and intensity of daily exposure. The questionnaire, which was designed to identify symptoms compatible with TMA-associated rhinitis, asthma, late respiratory systemic syndrome, and irritant responses, was administered to 18 of 20 workers. Total antibody binding was determined by the ammonium sulfate assay, and specific IgG and IgE binding was measured with an enzyme-linked immunosorbent assay (ELISA) to both PA-human serum albumin (PA-HSA) and TMA-human serum albumin (TM-HSA) conjugates. Specific IgE to TM-HSA was measured with the polystyrene-tube radioimmunoassay (PTRIA). The questionnaire identified nonirritant, immunologic symptoms caused by TMA primarily in the high-exposure group. Specific IgG, IgE, and total antibody binding to TM-HSA and PA-HSA were found exclusively in the high-exposure workers. One worker was identified with PA-associated rhinitis, high total antibody binding, and specific IgG and IgE to PA-HSA by means of the ELISA. The ELISA was as sensitive as the PTRIA in detecting the presence of specific IgE to TM-HSA. The present study demonstrates that an itemized questionnaire, combined with ELISA IgG and IgE determinations, may be a sensitive method for identifying workers who have or are at risk for developing TMA-induced respiratory symptoms.

Fractionation and biological characterization of Olea europea pollen extract.

Preliminary fractionation of Olea european pollen extract has been performed. At least 10 antigenic fractions have been found by crossed electrophoresis. After Sephadex gel filtration, two fractions with a molecular weight of 160,000 and 65,000 have been obtained. The fractions were evaluated for allergenic activity by two in vitro techniques: polystyrene tube radioimmunoassay (PTRIA) and basophil degranulation test (BDT), and by skin tests. All tests indicated that the most reactive fractions were those in the 65,000 molecular weight peak. BDT has been shown to be a very reliable method as compared with histamine release and other parameters. Although common antigenic fractions have been found for Lollium perenne and O. europea, no cross-allergenicity has been shown by PTRIA inhibition.

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.

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.

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 (PTRIA). 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 of toluene diisocyanate and HSA. Total serum antibody binding of 125I MDI-HSA was 15 μg/ml in worker A and 900 μg/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 industrial setting.

Quantitation and new antigenic determinant specificity of antibodies induced by inhalation of trimellitic anhydride in man.

We have studied the immune response of chemical workers who are exposed to trimellitic anhydride (TMA), a biologically reactive anhydride used in the manufacture of plastics. Serum antibody activity to TMA haptenized human serum albumin (TM-HSA) was measured in workers with three clinically defined respiratory tract syndromes; asthma-rhinitis, a late respiratory systemic syndrome (LRSS), and an irritant syndrome. IgE antibody binding of <sup>125</sup>I TM-HSA was quantitated by polystyrene tube radioimmunoassay and total serum antibody binding of <sup>125</sup>I-TM-HSA by an ammonium sulfate technique. IgE antibody activity ranged from 2 to 55 ng TM-HSA bound per milliliter and was associated with the asthma-rhinitis syndrome. Total serum antibody activity ranged from 0.4 to 56 <i>μ</i>g TM-HSA bound per milliliter and was associated with either the asthma-rhinitis syndrome or LRSS. Antibody specificity for TM-HSA was determined by inhibition studies using sodium trimellitate (NaTM) and a variety of TM substituted carriers. The hapten, NaTM, was an extremely poor inhibitor of IgE or total antibody binding of TM-HSA. The other TMA haptenized carriers were either noninhibitory or had to be added in milligram quantities to effect any inhibition as contrasted with complete inhibition achieved by microgram quantities of TM-HSA. These specificity studies indicate that the inhalation of TMA results in an antibody response with specificity for unique new antigenic determinants (NAD) which arise from the coupling of TMA with autologous respiratory tract proteins.

Serum IgE and IgG antibody activity against Aspergillus fumigatus as a diagnostic aid in allergic bronchopulmonary aspergillosis.

Serum IgE and IgG antibody activity against Aspergillus fumigatus was measured in 3 groups of subjects by 2 different immunologic methods. Group A consisted of 23 patients with allergic bronchopulmonary aspergillosis (ABPA). Group B was composed of 19 patients with extrinsic asthma who had marked immediate type skin reactivity to A. fumigatus (prick skin test, 3 or 4+) but no other manifestation of ABPA. Group C, the control group, was composed of 12 healthy subjects. Two immunological methods, including a solid-phase polystyrene tube radioimmunoassay and an iodine-125-labeled, A. fumigatus antigen radioimmunoassay, were used to study each patient's serum sample, so as to demonstrate IgE antibody activity against A. fumigatus (IgE-Af) and IgG antibody activity against A. fumigatus (IgG-Af). Both IgE-Af and IgG-Af were significantly greater among patients in Group A than among those in Group B and Group C, as measured by both methods (P is less than 0.001). The results of this study suggest that either method can be used as a diagnostic aid for ABPA. These methods may provide a laboratory test permitting diagnosis of ABPA in its early stages before bronchial or pulmonary destruction occurs.

Studies of perennial ragweed immunotherapy: Associated changes in cellular responsiveness, total serum antigen-binding capacity, and specific IgE antibody concentrations

Perennial ragweed immunotherapy was studied in 24 patients with ragweed pollenosis. Cellular responsiveness was determined by measuring the cellular reactivity and sensitivity to ragweed antigen E (RW-AgE) by RW-AgE-induced leukocyte histamine release. Serum blocking antibody content was determined by measuring the serum RW-AgE binding capacity by ammonium sulfate coprecipitation of bound RW-AgE. Specific IgE (anti-RW-AgE) concentration was determined by polystyrene tube radioimmunoassay. Cellular responsiveness decreased with continuing immunotherapy, as did the specific IgE concentrations. The serum RW-AgE binding capacity, in contrast, increased as treatment continued. The absence of demonstrable correlations between RW-AgE binding capacity and cellular reactivity, cellular sensitivity, and specific IgE concentrations contrasted impressively with the demonstration of multiple significant correlations between the change in the RW-AgE binding capacity and the other parameters studied. The degree of increase in RW-AgE binding capacity correlated significantly with the degree of decrease in both the specific IgE concentration (p < 0.04) and the cellular sensitivity to RW-AgE (p < 0.003). These findings suggest that the active process of blocking antibody production, rather than the passive presence of blocking antibody, is related to the process which decreases the specific IgE concentration and the degree of cellular responsiveness and therefore results in clinical improvement.