Methods: Binding of human IgE to anisakis and house dust mite proteins was investigated by immunoblot with serum from individuals sensitized to anisakis or shrimp. IgE binding patterns in the immunoblots were used for the identification of major Anisakis allergens, which were analysed by mass spectrometry-based proteomics in ESI-Orbitrap, after separation on SDS-gel. Results: Four new allergen candidates were identified. The first identified allergen was enolase, which is related to the

more »

... h allergen enolase. The other allergens were Heat Shock Protein-70 (HSP 70), tubulin, and glutathion-S-transferase, which are also present as allergens in house dust mite. Conclusions: Here we describe the identification of 4 novel IgE binding allergens in anisakis. The allergens might explain IgE cross-reactivity between anisakis and house dust mite or cockroach. Background: One important point in cow's milk allergy (CMA) is to stablish predicitve factors in acquiring tolerance. The aim of this study is to evaluate if a ratio of milk components: alpha-lactalbumin (a), beta-lactoglobulin (b), casein (C) and whole milk (WM) can contribute to predict tolerance development. Methods: It was a retrospective study that included patients with previous diagnosis of CMA evaluated at 6 years old. CMA was defined as a positive double blind placebo-controlled food challenge, open challenge or confirmed anaphylaxis plus positive specific IgE to cow's milk (higher than 3.5 kU/L or positive skin prick test), and tolerance was defined as acceptance of cow milk without previous symptoms. Specific IgE analysis to WM, a, b, and C were performed through Immunocap (Phadia AB). Ratios of milk components and whole milk (a /WM, b/WM, C/WM) were calculated and compared the results in 2 study groups: tolerant and persistent at 6 years old. Since values from both a /WM and b/WM ratios didn't follow a normal distribution, Mann-Whitney test was used to compare groups. For C/WM ratios, Student's t test was used as values were normally distributed. Results: It included 49 patients (27 male/22 female), 24 tolerant and 25 persistent. Average age of Immunocap test was 2.7 years (SD ¼ 1.4). Comparing the results from 2 groups, persistent and tolerant it was detected: no difference in a /WM (P ¼ 0.055, Mann-Whitney test), higher levels of b/WM in persistent group (P ¼ 0.023, Mann-Whitney test) and also higher levels of C/WM in persistent group (P ¼ 0.004, Student's t test). Conclusions: Higher ratios involving beta-lactoglobulin or casein components were detected in persistent patients. Thus, evaluating these markers precociously can be helpful in predicting CMA evolution.