Scope: β‐lactoglobulin (BLG) is a major cow milk allergen encountered by the immune system of infants fed with milk‐based formulas. To determine the effect of processing on immunogenicity of BLG, this article characterized how heated and glycated BLG are recognized and internalized by APCs. Also, the effect of heat‐induced structural changes as well as gastrointestinal digestion on immunogenicity of BLG is evaluated.Methods and results: The binding and uptake of BLG from raw cow milk and heated either alone (BLG‐H) or with lactose/glucose (BLG‐Lac and BLG‐Glu) to the receptors present on APCs are analyzed by ELISA and cell‐binding assays. Heated and glycated BLG is internalized via galectin‐3 (Gal‐3)and scavenger receptors (CD36 and SR‐AI) while binding to the receptor for advanced glycation end products (R AGE) does not cause internalization. Receptor affinity of BLG is dependent on increased hydrophobicity, β‐sheet exposure and aggregation. Digested glycated BLG maintained binding to sRAGE and Gal‐3 but not to CD36 and SR‐AI, and is detected on the surface of APCs. This suggests a mechanism via which digested glycated BLG may trigger innate (via RAGE) and adaptive immunity (via Gal‐3).Conclusions: This study defines structural characteristics of heated and glycated BLG determining its interaction with APCs via specific receptors thus revealing enhanced immunogenicity of glycated versus heated BLG.

Bovine immunoglobulin G (bIgG) was previously shown to enhance innate immune responses to toll-like receptor (TLR) stimulation, via induction of trained immunity. In this study, we investigated whether minimally processed dairy streams with high levels of whey proteins as potential infant nutrition ingredients could also induce trained immunity, and to what extent this can be explained by the presence of bIgG. The minimally processed whey ingredients serum protein concentrate (SPC) and whey protein concentrate (WPC) were tested for their ability to induce trained immunity in human peripheral blood monocytes. Both ingredients induced trained immunity as evidenced by an increased production of TNF-α and, to a lesser extent, of IL-6 upon stimulation with TLR ligands. This was comparable to isolated bovine immunoglobulin G (bIgG) that served as positive control. Depletion of bIgG from both whey protein-containing ingredients did not significantly inhibit the induction of trained immunity, suggesting that the streams contain other components in addition to bIgG that are able to induce trained immunity. These results indicate that minimally processed whey ingredients may contribute to protection against infections through enhancing innate immune responsiveness to pathogens.