Allergy diagnosis with the radioallergosorbent test: A comparison with the results of skin and provocation tests in an unselected group of children with asthma and hay fever.
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The ability to covalently couple Alternaria allergens to microcrystalline cellulose particles has permitted not only the measurement of IgE antibodies to Alternaria in patient serums but also the identification of allergenic fractions from crude Alternaria extracts. Crude aqueous Alternaria extracts from 3 commerical suppliers were coupled to cellulose but failed to bind more than 5% of total radioactive counts (TRC) when reacted with serums from highly sensitive patients. Fractionation of a commercial extract through Sephadex G-25 showed that almost all allergenic activity was located in a protein- and carbohydrate-containing peak eluting at the column void volume. These fractions were pooled and coupled to cellulose to yield a RAST polymer which produced up to 20% TRC binding when tested with serums from over 100 Alternaria-sensitive patients, and only up to 1% TRC binding with 17 nonallergic serums. The study of commercial Alternaria extracts by chromatographic and Rast inhibition techniques showed that present extracts are neither qualitatively or quantitatively comparable.
Seventy-nine patients with acute or former reactions to penicillin were investigated by a benzylpenicilloyl (BPO)-specific RAST and/or by skin tests with penicilloyl-polylysine (PPL), benzylpenicillin and penicilloic acid and the results were correlated with the different clinical pictures. Positive RAST and skin test results could be found in patients with anaphylactic shock, urticaria and serum sickness-like reaction and sometimes in a special group of exanthems, which are characterized by the existence of many different lesions at the same time, therefore called 'polymorphic exanthems', and often observed after high-dosage penicillin therapy. In cases of scarlatiniform or morbilliform exanthems no positive results were found. The BPO-specific RAST showed an overall correlation of 95-I% with skin tests using PPL. However, some patients with positive skin tests to benzylpenicillin and penicilloic acid did no have detectable circulating IgE antibodies to BPO. This emphasizes the need for including these antigens in in vitro methods. The RAST was informative even at the allergic reaction or in the first 15 days afterwards and seems to be very valuable for early diagnosis of penicillin allergy especially in cases when many drugs have been given.
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Several components in house dust extract can bind IgE, these interactions are not all immunologically specific. Non-immune interactions may conceivably contribute to the biological effects of house dust extract, they are actually found to interfere with the assay of allergen-specific IgE, especially when the serum under investigation has a high IgE content. By modification of the incubation medium, non-immune interactions can to a large extent be prevented. The immunological interactions between IgE and components in house dust extract have been investigated using twenty extracts, both in inhibition-type assay and in direct insolubilization assay. D. pteronyssinus-related allergens were found to be present in almost all extracts, however, not all house dust specific IgE reacted with mite-related allergens. Whether standardization of house dust extract is at all feasible, and if so, worthwhile, remains to be seen. It is clear, however, that no single figure can represent a completely reliable measure for the biological potency of a house dust extract.
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Immunoglobulin E-rich fractions and IgE-depleted fractions were prepared from individual and pooled sera of patients allergic to ragweed. The IgE-depleted fractions contained antibodies to ragweed associated with the immunoglobulins G, A and M. The IgE-rich fraction contained only IgE antibodies to ragweed, although other immunoglobulins were present in this fraction. The amount of antibodies in these fractions was determined by the radioimmunoassay described in a preceding publication. The inhibitory role of the non-IgE antibodies was examined in an allergosorbent test (RAST) by using various concentrations of the IgE-rich fraction. The RAST was performed using ragweed-coupled cellulose powder as well as paper discs. The IgE antibodies in concentration from 0.017 to 0.069 mug/ml were inhibited by 30-60% in the cellulose RAST by non-IgE antibodies in concentrations from 6.25 to 12.5 mug/ml. The inhibition effect was substantially less pronounced in the paper disc RAST. This would indicate that for diagnostic purposes, especially for examination of sera with high levels of IgG antibodies, it is more reliable to use the disc RAST instead of the cellulose powder RAST.
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