Sensitizing Capacity and Residual Allergenicity of Hydrolyzed Cow’s Milk Formulae: Results from a Murine Model

Abstract

Background: Cow’s milk allergy is the most common cause of clinically relevant adverse reactions to food in infants and children. Partially and extensively hydrolyzed formulae are used for the therapy and prevention of cow’s milk allergy. However, the immunogenic potency of hydrolyzed cow’s milk formulae to induce and/or enhance the allergic phenotype in vivo is still under debate. The aim of this study was to assess the sensitizing capacity and residual allergenicity of various partially and extensively hydrolyzed cow’s milk formulae in a murine model of cow’s milk allergy. Methods: BALB/c mice were immunized with either a cow’s milk formula or various partially and extensively hydrolyzed formulae. Immediate cutaneous hypersensitivity reactions and allergen-specific antibody production were assessed. Results: Although immunization with cow’s milk resulted in 12/13 cases in a positive skin test response to intradermal injection of cow’s milk formulae, only 1 mouse showed a positive skin test to one of the partially hydrolyzed formulae, and none showed positive reactions to other partially hydrolyzed formulae, any of the extensively hydrolyzed formulae, phosphate-buffered saline or the amino acid formula. However, 6 of 8 mice showed positive skin tests when immunized with partially hydrolyzed formulae and with one of the extensively hydrolyzed formulae. Conclusions: The residual allergenic potential is markedly reduced in many hydrolyzed formulae, but most of the formulae investigated could induce an allergic immune response in BALB/c mice. Our murine model seems to be suitable to investigate the sensitizing capacity of hydrolyzed formulae and to differentiate even between extensively hydrolyzed formulae.

Partially hydrolyzed whey proteins prevent clinical symptoms in a cow's milk allergy mouse model and enhance regulatory T and B cell frequencies

“…This finding corresponds with previous experiments in the same and other mouse models using partially hydrolyzed whey proteins [12,33,34]. Interestingly, despite the absence of clinical symptoms, we found a significantly increased whey-specific IgE response in whey hydrolysate sensitized animals.…”

Enzymatic Treatment of Whey Proteins in Cow’s Milk Results in Differential Inhibition of IgE-Mediated Mast Cell Activation Compared to T-Cell Activation

“…To date, different results with different ranges of CM protein hydrolysis have been reported in rodent models [21,25,26,27] and clinical studies [28,29,30,31]. In prevention strategies, there is not really a need to check for IgE binding, because specific IgE antibodies have not yet been produced.…”

A Protein’s Conformational Stability Is an Immunologically Dominant Factor: Evidence That Free-Energy Barriers for Protein Unfolding Limit the Immunogenicity of Foreign Proteins

“…Blood samples were drawn from the orbital sinus into capillary tubes every week after immunization. The immunization experiments using Phl p 7 were performed according to the methods of Niggemann et al (27), with slight modification. Five mice were injected i.p.…”

“…To examine whether a hyperstable protein, M5C mutant Phl p 7, was associated with a decrease in IgG production, BALB/c mice were immunized with M5C mutant Phl p 7 and wild-type Phl p 7 in aluminum adjuvant and were bled every week, according to previously described methods with slight modifications (27). The reactivities of these antisera to M5C mutant Phl p 7 and wild-type Phl p 7 were analyzed using ELISA.…”