Peanut protein allergens: the effect of roasting on solubility and allergenicity.

BACKGROUND: A contributing factor to food allergen stability is heat resistance. Peanut allergens in particular are resistant to heat, which results in their decreased solubility upon routine extraction and may have a profound influence on their continued presence in the digestive tract. Although there have been a number of studies characterizing soluble extracts of raw and roasted proteins, the relative solubility of the insoluble material following routine extraction for residual allergen characterization has not been investigated. The effects of various treatments on the re-solubilization and subsequent allergenicity of this insoluble peanut protein material are presented here. METHODS: Various methods to resolubilize the insoluble protein material were used, including pH, proteases and glycosidases. Protease digestion of nonextractable peanut proteins with pepsin, chymotrypsin and trypsin was performed in appropriate buffers as previously optimized for peanut proteins. Glycosidase activity in the presence of protease inhibitors was performed at pH 2. Digested samples were then subjected to SDS-PAGE/Western blot analysis using serum IgE from peanut-sensitive individuals. RESULTS: Progressive roasting of peanuts resulted in a significant decrease in protein solubility. The acidic proteins were resolubilized moderately at high pH, with solubility decreasing as pH approached the pI of the protein. However, at pH 2 the solubility increased dramatically. More extensive resolubilzation was observed with amylase treatment, presumably due to cleavage of glycoside of glycoproteins. The protein released into solution had a high IgE-binding capacity. While amylase was effective at resolubilizing this material, digestive tract proteases were not. CONCLUSION: The presence of these insolubilized peanut proteins provides a continuous source of major allergens to the gastrointestinal mucosal immune system.

A neonatal swine model for peanut allergy.

BACKGROUND: Peanut allergy represents a significant health threat in the United States. The factors contributing to the severity of the allergic response and the immunopathogenic mechanisms underlying peanut allergy remain to be completely characterized. As yet, no animal model has been developed that will completely mimic the physical, immunologic, and histologic features of food allergy. OBJECTIVE: The purpose of this investigation was to develop a neonatal pig model of peanut allergy that would mimic the allergic symptoms and the immunologic and histologic profile of human peanut allergy. METHODS: Newborn piglets sensitized intraperitoneally with peanut extract and cholera toxin were orally challenged repeatedly with peanut meal. Physical symptoms, including emesis, lethargy, diarrhea, and respiratory distress, were monitored to determine the allergic response. Immunologic assessment was conducted through use of skin testing and the antigenic response to peanut proteins. Histologically, tissues derived from the esophagus, stomach, small intestine, and colon were assessed for morphologic changes after the oral challenge. RESULTS: Peanut-sensitized pigs responded with physical symptoms that mimicked those seen in double-blinded, placebo-controlled oral food challenges to peanuts in children and adults. Skin testing suggested an IgE-mediated response; this was confirmed by a negative passive cutaneous anaphylaxis response of heat-treated sera obtained from peanut-sensitized animals. Damage to villi of the small intestine was similar to that seen in endoscopically obtained tissue specimens from certain food-allergic individuals. CONCLUSION: The neonatal pig model of peanut allergy mimics the physical and immunologic characteristics of peanut allergy in human beings. The model will be useful for determining IgE-mediated mechanisms and conducting endoscopic histologic assessment of tissues and immunotherapeutic intervention strategies with repeated allergen challenges.