Alanine Scanning of the Unstructured Region of Ara h 2 and of a Related Mimotope Reveals Critical Amino Acids for IgE Binding.

SCOPE: The unstructured region of Ara h 2, referred to as epitope 3, contains a repeated motif, DYPSh (h = hydroxyproline) that is important for IgE binding. METHODS AND RESULTS: IgE binding assays to 20mer and shorter peptides of epitope 3, defines a 16mer core sequence containing one copy of the DPYSh motif, DEDSYERDPYShSQDP. This study performs alanine scanning of this and a related 12mer mimotope, LLDPYAhRAWTK. IgE binding, using a pool of 10 sera and with individual sera, is greatly reduced when alanine is substituted for aspartate at position 8 (D8; p < 0.01), tyrosine at position 10 (Y10; p < 0.01), and hydroxyproline at position 12 (h12; p < 0.001). IgE binding to alanine-substituted peptides of a mimotope containing the DPY_h motif confirm the critical importance of Y (p < 0.01) and h (p < 0.01), but not D. Molecular modeling of the core and mimotope suggests an h-dependent conformational basis for the recognition of these sequences by polyclonal IgE. CONCLUSIONS: IgE from pooled sera and individual sera differentially bound amino acids throughout the sequences of Epitope 3 and its mimotope, with Y10 and h12 being most important for all sera. These results are highly significant for designing hypoallergenic forms of Ara h 2.

Role of Food Allergy Education: Measuring Teacher Knowledge, Attitudes, and Beliefs.

INTRODUCTION: Almost 6 million children suffer from food allergies with roughly 2 affected per classroom. Deficiencies in knowledge and discrepancies in attitudes within school staff when addressing food allergies are associated with barriers to care. In this study, school teacher knowledge, beliefs, and attitudes were measured before and after a food allergy educational session. METHODS: Three hundred seventy-five personnel of similar age, socioeconomic status, ethnicity, and educational level completed the Chicago Food Allergy Research survey before and after a 1-hour educational session in 6 private schools in Houston randomly assigned into an intervention (n = 4) and control group (n = 2). Responses were measured using frequencies and percentages. The group score differences and survey question comparisons were evaluated with a linear mixed-effects model. RESULTS: Posttest, the intervention group had knowledge scores 19.58% points higher than control (95% confidence interval = 16.62-22.53; P < .001) with no differences pretest. Odds of agreeing that injectable epinephrine is important was higher in the intervention schools posteducation. Within the intervention group, personnel were more likely to agree to injectable epinephrine use for children posteducation. CONCLUSION: A 1-hour educational session improved knowledge and attitudes in personnel in the intervention schools. Given the growing prevalence of food allergy, the emphasis on food allergy education is crucial to allow for familiarization of the condition, early recognition of anaphylaxis, and promotion of injectable epinephrine use.