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Background The present study investigated to what extent shrimp allergic individuals were IgE-sensitized to anisakis, crab and house dust mite and whether tropomyosin was responsible for IgE cross-reactivity. Methods 29 Individuals with self reported shrimp allergy were recruited by advertisements in local and national news-papers in Norway. Anamnesis was taken, skin prick tests (SPT) were performed and positive responders to shrimp were studied further with basophile activation test (BAT), ImmunoCAP analyses and western blotting. Results Of the 29 persons studied, 10 (34%) had positive SPT against shrimp and house dust mite, 9 (31%) against shrimp tropomyosin and 3 (10%) against anisakis. Individuals with positive SPT to shrimp all showed positive basophilic responses to house dust mite, while 43% responded to shrimp, 25% to anisakis and 36% to crab in BAT. Moreover, SPT, BAT as well as ImmunoCAP analyses showed a positive correlation of IgE-reactivity between anisakis and shrimp, house dust mite and crab. Immunoblot studies indicated that these responses are not completely explained by cross-reactivity towards tropomyosin. Conclusions The current study indicates a positive correlation between IgE-mediated reactions to shrimp, anisakis, house dust mite and crab, which may not be completely explained by cross-reactivity against tropomyosin.

Visualization by autoradiography of specific IgE binding in crossed radioimmunoelectrophoresis (CRIE) and other 125I-immunoautoradiography (IAR) techniques is done in two different ways; either by traditional direct autoradiography (D-ARG), where the film is exposed to the 125I-anti-IgE incubated sample at room temperature, or by indirect autoradiography (ID-ARG), applying intensifying screen, low-temperature exposure and, eventually, pre-exposure. This study confirmed that D-ARG provided the benefits of simplicity and better image resolution with the disadvantage of prolonged exposure periods. ID-ARG reduced the exposures needed to produce film image densities of 0.01 and 0.1 A540 nm (i.e. autoradiographic sensitivity (AR sigma) and autoradiographic speed (ARs] to 1/18 and 1/55 respectively of the corresponding exposures in D-ARG. The lowest detection limits for 125I in 24 h were 1.2 cpm mm-2 with the indirect and 6.8 cpm mm-2 with the direct systems investigated. The major drawbacks of ID-ARG were inferior image resolution and higher background levels, especially when pre-exposure was included.

The effects of enzymic cleavage and perturbing the conformation of the allergenic and antigenic determinants of hens egg white albumin (OA) were examined. Hens egg white extract of a total protein concentration 8.43 g/l was prepared. Isoelectric focusing in sodium docedyl sulfate and polyacrylamide gel peptide maps for the crude egg white extract showed 26 spots visualized by staining with Coomassie blue. The OA was purified using a TSK-2000 gel filtration chromatography column. The specific allergenic reactivity of the purified·A as measured by RAST inhibition and direct RAST was relatively high: 3 μg gave an inhibition of approximately 10%.The cleavage of OA with cyanogen bromide resulted in 4 fractions, all capable of binding specific IgE with the first peak showing the highest inhibition. Thermal denaturation of OA had no direct effect on the antigenic reactivity. RAST inhibition values for the denatured protein were similar to those of the native protein. Carboxymethylation of OA gave a product with only 20% of the inhibition reactivity. Further treatment with trypsin did not abolish the allergenic and antigenic reactivities as shown by RAST inhibition and by deflection of OA line in rocket line immunoelectrophoresis. On the other hand, limited pepsin hydrolysis destroyed the antigenic structure of the molecule. The reactivity of OA is thus relatively stable and could easily be retained making it possible to identify the allergenic determinants of enzymic hydrolysates used for elucidating the antigenic structure of the molecule.

The CD-calcium binding loop of Allergen M encompasses the two homologous tetrapep-tides Asp-Glu-Asp-Lys and Asp-Glu-Leu-Lys. These tetrapeptides were recently suggested to be mutually critical for the immunological specificity of this region. To obtain evidence of the validity of this hypothesis, four peptide analogues encompassing these tetrapeptides at the termini were produced by manual solid-phase peptide synthesis. The structures of these peptides were ascertained by amino acid analyses and end-group sequence determination. The crude preparations were prepared by high-performance liquid chromatography and high-voltage electrophoresis. The immunological reactivity was investigated by using the preliminary purified peptides in the IgE and IgG test systems used. The results of radioallergosorbent test inhibition showed that all the synthetic peptides encompassing a minimum of two of the tetrapeptides have the capacity to bind specific IgE of the serum pool. The composition and sequence of the six spacer amino acids between these tetrapeptides were of no importance for the acitivity. The highest inhibition percentage was obtained by the peptides derived from the primary structure of Allergen M. No activity was found for the control peptides synthesized under identical conditions. Similar results were shown by PIC tests, using two cod-allergic sera and two recipients. In the IgG test system, all the four peptides containing the two terminal tetrapeptides showed an antigenic reactivity. They deflected allergen M line in rocket line immunoelectrophoresis. No deflection was given by the control peptides or proteins used. We conclude that the immunological reactivity of region 41–64 of allergen M is determined by 3 homologous tetrapeptides, repeated in three sites interspaced by six amino acids in a segment of 24 residues.

Allergens in birch (Betula) pollens from B. pendula grown in Australia and Norway, B. davurica and B. populofolia and from alder (Alnus incana) were identified by electroblotting, following separation by SDS-PAGE, transfer to nitrocellulose membranes and incubation with sera from birch pollenallergic subjects. Of 42 antigenic components detected by protein staining in the pollen extract from B. pendula grown in Norway, 17 bound IgE. The allergenic components included those already reported in the literature at MWs of 40, 29, 25, 17 and 10–12 kd, as well as previously undescribed components at MWs of 90, 79, 60, 50, 38, 35, 31, 27, 23, 16, 15 and 14 kd. The major IgE-binding components were located in the low MW region 10–17 kd for all species of birch and alder pollen proteins studied. Results reported here provide the first evidence of birch and alder pollen allergies in Australia. Extensive heterogeneity was observed amongst the sera from birch pollen-allergic subjects in both Norway and Australia. Cross-reactivity appears to exist among the proteins present in pollen from the various birch species and from alder.

Background Bronchial hyperresponsiveness (BHR) is common in bakery workers. The relation between bronchial responsiveness measured with a tidal breathing method and smoking, airway symptoms, IgE-sensitization, nasal indices of inflammation and flour dust exposure have been studied with bronchial responsiveness expressed as a continuous outcome. Material and methods Bakery workers (n = 197) were subjected to interviews, questionnaires, allergy tests, workplace dust measurements and bronchial metacholine provocation. Eosinophil cationic protein (ECP) and alpha(2)-macroglobulin were measured in nasal lavage. Bronchial responsiveness was expressed as slope(conc), a measurement based on regressing the per cent reduction in FEV1 at each provocation step. Results BHR expressed as slope(conc) was associated with smoking (P = 0.009), asthma symptoms at work (P = 0.001), and occupational IgE sensitization (P = 0.048). After adjusting for baseline lung function the association between BHR and IgE sensitization was no longer present. We demonstrated an association between nasal ECP and BHR (slope(conc) < 3: P = 0.012), but not to alpha(2)-macroglobulin in nasal lavage. No association was seen between BHR and current exposure level of flour dust, number of working years in a bakery or a history of dough-making. Conclusions BHR is related to baseline lung function, work-related asthma symptoms, smoking and nasal eosinophil activity, but not to occupational IgE sensitization and current flour dust exposure when measured with metacholine provocation. The slope(conc) expression seems to be a useful continuous outcome in bronchial responsiveness testing. (Less)

The use of different diagnostic criteria has considerable consequences for the prevalence estimates of occupational rhinitis. There is a strong relationship between occupational rhinitis and lower airway symptoms. Storage mites appear to be important occupational allergens in Norwegian bakeries.To study the consequences of various diagnostic criteria on the prevalence of occupational rhinitis, assess the prevalence of IgE sensitization and explore the relationships between upper and lower airway symptoms and between symptoms and IgE sensitization.A total of 197 employees in 6 bakeries were interviewed and completed a questionnaire. A skin prick test was performed, total and specific IgE were determined and a histamine release test was performed for relevant allergens. The criteria for the diagnosis of occupational rhinitis were based on the 1994 International Consensus Report on Rhinitis.The prevalence of occupational rhinitis varied between 23% and 50%, depending on the criteria used. The occurrence of nasal symptoms was found to precede the development of lower airway symptoms. Occupational rhinitis, both IgE- and non-IgE-mediated, was associated with asthma symptoms. The most frequent causes of sensitization (20%) were different species of storage mites. Storage mite sensitization was related to occupational rhinitis and work exposure.