Detection of antibodies to proteases used in laundry detergents by the radioallergosorbent test.

Two proteases, Esperase and Alcalase, derived from Bacillus licheniformis and B. subtilis, respectively, are used in laundry products. In testing for the prevalence of IgE antibodies to these enzymes in sera among 300 laundry product workers, we experienced two problems in the establishment of a reliable RAST for these antigens. The first problem was the propensity of the allergen, Esperase, to undergo autolysis, suggesting that solid-phase Esperase might also lose reactivity through degradation. Treatment of Esperase with phenylmethylsulfonyl fluoride stabilized the enzyme and permitted the synthesis of a stable solid-phase antigen. The second problem was the finding that sera reactive with Esperase in the RAST were also reactive with Savinase, an enzyme from B. licheniformis to which the workers were not exposed. Immunochemical analyses of the three enzymes with specific rabbit antisera by gel diffusion and by two-site immunoradiometric assay demonstrated that they were not cross contaminated to any appreciable extent. RAST inhibition demonstrated that solid-phase Esperase possessed unique allergenic determinants in that the reactivity of IgE antibodies was inhibited by low concentrations of Esperase and only by very high concentrations of Alcalase and Savinase. In contrast, the reactivity of solid-phase Alcalase was occasionally inhibited equally well by Esperase and Alcalase. Most strikingly, the reaction of IgE antibodies with solid-phase Savinase was always inhibited by comparable quantities of Esperase, Alcalase, and Savinase. Thus, the establishment of the RAST for these proteases appears to require the use of phenylmethylsulfonyl fluoride to retard autolysis, and the results must be interpreted with caution because IgE antibodies in certain sera demonstrate cross-reactivity with Alcalase and Savinase.

Facial edema and eosinophilia. Evidence for eosinophil degranulation.

Two patients had recurrent facial edema and peripheral blood eosinophilia. One patient showed a marked increase in the serum level of the eosinophil granule major basic protein. In both patients, skin biopsy samples showed nonspecific mononuclear cell inflammation with few eosinophils. However, immunofluorescence staining showed extracellular localization of the major basic protein within the dermis, similar to that previously shown in chronic urticaria and the recently described syndrome of episodic angioedema with eosinophilia. These observations provide further evidence that degranulation of eosinophils occurs in the skin and suggest that eosinophil mediators may play a role in the development of cutaneous edema.

[Immediate hypersensitivity to quinine]. / Hypersensibilité immédiate à la quinine.

Systemic anaphylactic reaction with shock was observed after ingestion of an antipyretic combination product containing quinine. That quinine was responsible for the reaction was proven by immediate skin tests and by oral challenge tests. The demonstration of specific IgE's by a radioallergosorbent test (RAST) provided evidence for an immediate hypersensitivity mechanism.

Aspirin intolerance and asthmal induction of a tolerance and long-term monitoring.

Acetylsalicylic-acid (ASA) intolerance is well recognized as a possible cause for exacerbating asthma. It has been postulated that if this could be overcome, long-term aspirin administration could improve asthma symptoms and enable reduction of the use of other anti-asthmatic drugs. We succeeded in inducing an ASA tolerance in nine corticosteroid-dependent asthmatics, and this tolerance lasted at least 1 month and at most 1 year. Progressive deterioration in lung function was seen, however, in all patients taking ASA for more than 1 month, unless the corticosteroid intake was increased. These results suggest that ASA long-term treatment is of no help to severe corticosteroid-dependent, ASA-sensitive asthmatics. This conclusion differs from other reports on ASA intolerance in asthma.

Charcot-Leyden crystal protein and eosinophil granule major basic protein in sputum of patients with respiratory diseases.

Sputum eosinophilia is characteristic of bronchial asthma, and sputum specimens from patients with asthma contain eosinophil-derived substances including Charcot-Leyden crystals (CLC) and the eosinophil granule major basic protein (MBP). Prior observations have indicated that an elevated sputum level of MBP is associated with asthma. To determine whether CLC protein was also elevated in asthma, we measured by specific radioimmunoassays the quantities of MBP and CLC protein in sputum specimens from 106 consecutive patients with various respiratory diseases and in sputum specimens from 10 patients hospitalized for asthma. The CLC protein was detected in all sputum samples, and the levels in the 116 samples averaged 3.5 micrograms/ml (range, 0.01 to 25 micrograms/ml). The CLC protein was significantly elevated in sputum specimens from patients with acute asthma and from patients with certain other respiratory diseases when associated with bronchopulmonary infection. In contrast, MBP levels in the 116 samples averaged 0.32 micrograms/ml (range, 0.01 to 8.8 micrograms/ml) and were significantly elevated only in patients with asymptomatic or acute asthma. Sputum MBP was not elevated in patients with bronchopulmonary infections unless acute asthma was also present. Thus, an elevated sputum MBP level was specifically associated with asthma, whereas an elevated CLC protein level was associated with both asthma and with bronchopulmonary infection in certain patient groups.

Induction of late cutaneous reaction by kallikrein injection: comparison with allergic-like late response to compound 48/80.

Although IgE antibody is able to provoke a late cutaneous allergic response (LCAR), the mechanism of its development is not clear. It seems to involve vasoactive and clotting mediators released from mast cells and basophils. Substances biologically similar to plasma kallikrein (KK), a Hageman factor activator, have been shown to be released by basophils in the course of an IgE-dependent reaction. Because compound 48/80 induces LCAR-like responses in the skin, we compared the ability of tissue KK and compound 48/80 to induce late cutaneous reactions (LCRs). All 40 test subjects showed an immediate wheal and flare reaction (WF) after both KK and 48/80. The WF was followed by an LCR characterized by diffuse edema, pain, and erythema in 36 of 40 subjects with 48/80 and 26 of 40 with KK. The LCRs evoked by the two agents did not differ in their appearance or in their time of development. These reactions increased until the 5 hr mark, began to decrease at the 10 hr mark, and were gone after 24 hr. Histologic study showed edema and a mixed cell infiltration with fibrin deposition. Challenge of the original injection site and a new site after 1 or 2 wk showed a local refractory state lasting 2 wk. Prednisone almost totally suppressed the LCRs induced by both KK and 48/80.

[Radioimmunologic assay of the major basic protein of eosinophilic granules in sputum. A diagnostic contribution in asthma]. / Dosage radioimmunologique de la protéine majeure basique du granule éosinophile dans l'expectoration. Un appoint diagnostique dans l'asthme.

The concentration of major basic protein (MBP) of the eosinophilic granules in bronchial expectoration (Sputum) were measured by radioimmunoassay in 204 subjects suffering from various illnesses of both the respiratory tract and other systems. 31 of these subjects were later revealed to have asthma and all had very high sputum MBP levels. The MBP level was much greater in asthmatics than any other respiratory or non-respiratory ailment and in a statistically significant fashion (p less than 0.001). 25 other subjects who were in hospital for exacerbations of asthma were subsequently tested; their MBP concentrations were higher still. During the course of their hospital stay the sputum MBP levels fell significantly following treatment with steroids and broncho-dilators. The measurement of the MBP level would seem useful in those situations where the diagnosis of asthma is not evident from the history and may help in predicting the effectiveness of steroid treatment.