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PubMed1978

Studies of the radioallergosorbent test and the lymphocyte stimulation index in symptomatic ragweed pollenosis.

We followed 47 ragweed hay fever patients (not on immunotherapy) through a ragweed season by means of clinical evaluation and symptom scores. The radioallergosorbent test (RAST) and lymphocyte stimulation index (LSI) were done to determine their reliability as predictors of clinical symptomatology. Symptom scores of 10 or above, denoting at least moderate daily symptoms, occurred in 36 patients (80%). Preseasonally, RAST class 2 or above was found in 34 patients (75%), of whom 29 patients (64%) had symptom scores of 10 or above. At that time, LSI was elevated in about 50% of the patients. Patients in RAST class 3 had the highest LSI as a group. Seasonal exposure boosted the LSI in 50% of the patients, whereas 25% were unchanged and 25% showed a decrease. The results suggest that neither RAST nor LSI are reliable predictors of clinical severity in ragweed pollenosis.

Adult↗
PubMed1976

Radioallergosorbent test (RAST) studies with insolubilized polysaccharides.

Polysaccharide preparations from Aspergillus fumigatus and Candida albicans when used with some human sera were found to fix complement and precipitate with specific antibodies present in the sera. Precipitins consisted predominantly of IgG with traces of IgM and IgA. Following insolubilization of the fungal polysaccharide preparations with the N-carboxyanhydride of L-leucine, reactivity with serum precipitins was retained and reaction with IgE antibodies was demonstrated using the radioallergosorbent test (RAST). Results showed that for A. fumigatus and C. albicans at least, polysaccharide-specific IgE antibodies probably occur in sera from some hypersensitive individuals. We suggest that chemical methods of the type described which induce polymerization or cross-linking will be useful for detecting IgE antibodies in the RAST. Such an approach could be used with a variety of polysaccharide preparations which cannot be coupled to polysaccharide solid supports by the usual methods.

Antibodies, Fungal↗
PubMed1985

Serum immunoglobulin E levels in onchocerciasis: the development of a radioallergosorbent test for Onchocerca volvulus infection.

The effects of Onchocerca volvulus infection on immunoglobulin levels have been studied in subjects from three localities with different onchocerciasis prevalence rates. Infestation by O. volvulus provoked a 200-500% rise in total serum IgE levels. Immunoglobulins G, M. and A were increased less dramatically. To study further the IgE response, radioallergosorbent test (RAST) systems were developed using total extract of O. volvulus and Onchocerca supernatant (excretory-secretory) antigens. Both antigen preparations reacted with more than 80% of the onchocerciasis sera from the hyperendemic village, but the reactions were not proportionate to skin microfilarial density. There were also positive reactions with sera of some individuals whose skin biopsies showed no microfilariae, indicating prior exposure to the parasite. European control sera tested under similar conditions reacted only slightly in the RAST system. The developed RAST systems may thus be used for determining prior exposure to O. volvulus.

Cameroon↗
PubMed1977

Lymphocyte response to grass pollen antigens: a correlation with radioallergosorbent test and effect of immunotherapy.

The deoxyribonucleic acid (DNA) synthetic response of cultured lymphocytes to grass pollen antigens was significantly higher in 66 untreated atopic patients than in 15 non-atopic subjects (p less than 0.05). There was no correlation between the DNA synthetic response and the specific IgE antibody level as measured by radioallergosorbent test in the 81 subjects (r= 0.13; p greater than 0.2). The effect of immunotherapy on proliferative lymphocyte response and specific IgE antibody response to grass pollen antigens was evaluated prospectively in 22 atopic patients. The DNA synthetic response to grass pollen antigens was significantly reduced after immunotherapy (p less than 0.01) but the specific IgE antibody level did not change significantly.

Antigens↗
PubMed1989

[Dermatophagoides farinae and Japanese cedar (sugi) pollen specific immunoglobuline E antibodies in asthmatic children measured by radioallergosorbent test and enzyme-linked immunosorbent assay].

In recent years, pollinosis caused by Japanese cedar pollen (Cryptomeria japonica, sugi in Japanese) has been increasing strikingly in Japan. To investigate the prevalence and age-related sensitization of sugi pollen in children, the specific IgE antibody was determined in 204 asthmatics (129 boys, 75 girls) by the enzyme-linked immunosorbent assay (ELISA) and radioallergosorbent test (RAST), and compared to Dermatophagoides farinae (D.f.). There was a good correlation in the total serum IgE levels measured by ELISA and PRIST. We think that the ELISA is a reliable and valuable method which uses no radioactive material in a massive epidemiological survey. D.f. specific antibody levels increased in early infancy and remained high, and there was no difference between ELISA and RAST score. The sugi-basic-protein specific antibody measured by ELISA was detected in early infancy, and increased slowly for 5 to 6 years reaching its highest level at 7 to 12 years followed by decreasing levels, as revealed by both ELISA and RAST. Infants born in spring, the sugi pollinating season, showed a lower level of sugi-specific IgE because they were exposed the pollen 6 months to one year later. We believe that sensitization of Dermatophagoides farinae in early infancy could influence subsequent sensitization by sugi pollen in later life.

Adolescent↗
PubMed1978

Demonstration of distinct allergens by means of immunological methods. Comparison of crossed radioimmunoelectrophoresis, radioallergosorbent test and in vivo passive transfer test.

An immunosorbent column was prepared containing a purified major allergen fraction from codfish (DS 22) covalently coupled to agarose. Sera from patients allergic to codfish were run through the column at pH 7.2. After extensive washing, the IgE retarded in the column was eluted with a buffer at pH2.5. The original sera and fractions from the chromatography experiments were examined by means of crossed immunoelectrophoresis (CIE), crossed radioimmunoelectrophoresis (CRIE), radioallergosorbent test (RAST) and in vivo passive transfer (PK) tests using the DS 22 from codfish and a crude codfish extract. The experiments demonstrated that the crude extract contained a minor codfish allergen (antigen-17-cod) which was distinct from DS 22. RAST was the most convenient technique for the identification of fractions containing allergenic activity. The PK tests served to prove the biological activity in vivo. CIE/CRIE were superior to RAST and PK tests regarding their ability to identify distinct allergens. Full agreement was found between results using different techniques including the immunosorbent experiments. Some of the radiostaining in CRIE, however, was misleading due to coprecipitation of DS 22 in several precipitates in the CIE preparations of the crude codfish extract.

Allergens↗
PubMed1986

The influence of antibody affinity on the radioallergosorbent test (RAST) and in vitro histamine release. Studies with hapten-specific monoclonal IgE antibodies.

Four murine monoclonal IgE antibodies specific for the hapten, 4-hydroxy-3-nitrophenylacetyl (NP), had been previously found to be heteroclitic in nature in that they bound the crossreacting hapten, 4-hydroxy-3-iodo-5-nitrophenylacetyl (NIP), with greater affinity than NP. The influence of antibody affinity on the results of two commonly used assays for IgE, namely the radioallergosorbent test (RAST) and histamine release from rat peritoneal mast cells, was studied using these antibodies. In general, in agreement with previous reports, it was found that affinity influences both RAST and histamine release; however, the affinity constants deduced from equilibrium dialysis measurements for the reactions with monovalent haptens were not directly related to the activities of the antibodies as reflected in assays using multivalent hapten protein conjugates.

Animals↗
PubMed2004

Immunoblot and radioallergosorbent test inhibition studies of allergenic cross-reactivity of the predatory mite Amblyseius cucumeris with the house dust mite Dermatophagoides pteronyssinus.

BACKGROUND: In 1999, an extensive study among bell pepper growers showed that a predatory mite, Amblyseius cucumeris, is a potentially relevant source of occupational allergens because 23% of the population had positive skin prick test reactions. OBJECTIVE: To investigate whether cross-reactivity between A. cucumeris and Dermatophagoides pteronyssinus is responsible for the cosensitization to both mite species found in 58.7% of A. cucumeris-sensitized greenhouse workers. METHODS: Fifteen serum samples from greenhouse workers with work-related inhalant allergy and a positive radioallergosorbent test (RAST) reaction to A. cucumeris or D. pteronyssinus were selected for immunoblot analysis using extracts of both mites. A subselection (n = 5) was used for RAST and immunoblot inhibition to investigate potential cross-reactivity. RESULTS: On immunoblot, 2 distinct patterns were observed: one pattern showed common protein bands in A. cucumeris and D. pteronyssinus blots suggestive of cross-reactivity between A. cucumeris and D. pteronyssinus and the other pattern showed no shared protein bands. Dermatophagoides pteronyssinus RAST inhibition with A. cucumeris extract was low in 4 serum samples (<25% inhibition) and nearly absent in 1 serum sample; A. cucumeris RAST inhibition with D. pteronyssinus extract was high in 1 serum sample (75% inhibition), low in 2 serum samples (35% and <15% inhibition), and absent in 2 serum samples. These results were confirmed by immunoblot inhibition experiments. CONCLUSIONS: Amblyseius cucumeris, a new occupational allergen, has species-specific antigens and common antigens that are cross-reactive with the house dust mite D. pteronyssinus.

Adult↗
PubMed2005

Hetastarch allergy and positive latex radioallergosorbent test in a patient suffering cardiovascular decompensation during multiple perioperative periods.

The cause of sudden cardiovascular collapse in the perioperative period can be elusive. Allergy may be overlooked as a cause. When allergy is considered, latex is often suspected. Because hetastarch is frequently used in situations involving hypovolemia and hypotension, and because allergic reactions to it are rare, it may be overlooked as a possible allergen. We report a case of a patient suffering cardiovascular decompensation during four nonconsecutive perioperative periods before it was determined that she was allergic to hetastarch. She also had a very highly positive latex radioallergosorbent test, suggesting a latex allergy.

Aged↗
PubMed1979

Standardization of radioallergosorbent test (RAST) and allergen extracts in terms of the WHO standard for IgE.

An 131I-labelled IgE myeloma, standardized against the WHO reference serum for IgE was bound in varying concentrations to discs bearing anti-human light chain. These were used to construct a calibration curve for the subsequent binding of 125I-labelled anti-IgE, thus relating 125I counts to WHO IgE units. The calibrated anti-IgE could then be used to measure the actual amount of specific IgE bound to an allergen disc in the radioallergosorbent test. Using this standardized system, soluble allergen extracts could then be assayed by their ability to inhibit the binding of a defined amount of specific IgE, thereby enabling their activity to be expressed in WHO IgE units.

Allergens↗
PubMed1980

The development of a radioallergosorbent test (RAST) for murine IgE antibodies.

A purified monoclonal IgE preparation, isolated from the ascitic fluid of mice bearing a hybridoma secreting IgE with specificity to ovalbumin, was used for the production of goat anti-murine IgE (GAME) antiserum, which was then rendered monospecific for the epsilon chain. Another monoclonal hybridoma IgE preparation with specificity for the 2,4-dinitrophenyl group was isolated from ascitic fluid in a relatively pure state by affinity chromatography and used in the form of an immunosorbent to isolate antibodies from the monospecific goat serum. The GAME antibodies were 125I-labeled and used to develop a radioallergosorbent test (RAST) for the quantitation of murine IgE antibodies specifically adsorbed onto antigen-coupled paper discs. The RAST was specific for antibodies of the IgE class only and was as sensitive as and more accurate than PCA assay. RAST results on sera of mice treated with tolerogenic conjugates indicated a reduction in the affinity and concentration of the IgE antibody populations on suppression of the IgE response. The effect of interference by non-IgE antibody populations on the RAST curves has been discussed.

Absorption↗
PubMed1989

Radioallergosorbent test (RAST) for specific IgE antibody to lidocaine, procaine and methylparaben.

Although anaphylactoid reactions to local anesthetics are well known, a radioallergosorbent test (RAST) to detect specific drug reagin (IgE) anti-body has not been developed. We established RAST for local anesthetics by using carboxylic acid derivatives of lidocaine, procaine and methylparaben. Serum samples were taken from 100 volunteers who were regarded to be nonallergic to the drugs used. Negative RAST values obtained from these volunteers were 1653 +/- 254(SD) cpm (lidocaine), 2750 +/- 264 cpm (procaine), and 2805 +/- 336 cpm (methyl paraben).

Journal Article↗
PubMed1976

Analysis of the potency of extracts of June grass pollen by their inhibitory capacities in the radioallergosorbent test.

The potencies of 11 commercial extracts of June grass pollen were analyzed by skin test end point titrations and compared to potencies as determined in vitro (1) by the radioallergosorbent test (RAST), (2) by Group I antigen content, and (3) by protein nitrogen units (PNU). RAST potencies were determined by the capacity of the extract to inhibit the binding of IgE antibody to solid-phase allergen, and they were expressed as the quantity of extract required for 50% inhibition of binding. Potencies determined by skin testing in 8 patients were significantly related among the various patients in 19 of 27 comparisons and showed differences of up to 95,000-fold in the strengths of the extracts. Estimation of potencies by RAST inhibition showed approximately a 100-fold difference among the extracts and in 5 of 8 cases these were significantly related to potencies measured by skin tests. Similarly, PNU determinations and Group I determinations were also significantly related to potencies by skin test titration in 5 of 8 and in 4 of 8 comparisons, respectively. Comparison of the geometric mean skin test potencies with RAST, PNU, and Group I potencies revealed that all were significantly related to skin test potencies although the correlation of RAST and skin potency was the highest. The results indicate that measurement of potency by RAST inhibition compares favorably with other in vitro measurements of potency. These results are compared with those of a prior study with extracts of short ragweed, and the reasons for the differences between the results in the two studies are discussed.

Absorption↗
PubMed1976

Detection of allergy to nuts by the radioallergosorbent test.

The diagnosis of food allergy is often difficult to make by conventional means. Histories are frequently ambiguous, and skin testing is of dubious reliability because of the number of false-positive and false-negative reactions. We have evaluated the radioallergosorbent test (RAST) for the in vitro measurement of the specific IgE antibodies to nuts, including Brazil nut, almond, walnut, pecan, cashew, and the legume, peanut. Serums were obtained from 18 patients with a history of nut allergy and IgE level and specific IgE antibodies were measured. Thirteen of the 18 patients had significantly elevated IgE antibody (greater than twice control) to one or more of the allergens. Prausnitz-Küstner tests on selected serums in general corroborated the results of the in vitro studies. Five patients had RAST elevations to 2 or more nuts. As a group RAST-positive patients had elevated mean serum IgE levels and more severe clinical symptoms (p less than 0.01). The specificity and cross-reactivity of IgE antibodies to different nut antigens was investigated by RAST inhibition with serums from 5 patients having high levels of IgE antibody. In 4 patients no cross-reactivity between Brazil nut and peanut was found. In contrast, several nut extracts inhibited the reaction of pecan allergen with IgE antibodies. These results indicate that specific IgE antibodies can be measured by RAST in patients with nut allergy and the cross-reactivity of nut antigens can be investigated. RAST would appear to be most useful in confirming the diagnosis of nut hypersensitivity in children or in highly allergic patients in whom skin testing poses a risk of anaphylaxis.

Absorption↗
PubMed1976

Serum immunoglobulin E concentrations and radioallergosorbent tests in children with atopic dermatitis.

Serum IgE concentrations and the presence of allergen-specific IgE were determined in a series of 23 children with atopic dermatitis. In this group 83% had significantly elevated serum levels of IgE, 91% had coexistent respiratory allergy, 70% had radioallergosorbent test (RAST) evidence of pollen hypersensitivity, and 43% gave a history and demonstrated a RAST score consistent with milk or egg hypersensitivity. In patients with eczema a significant proportion of the elevated serum IgE appears to be antigen specific.

Adolescent↗