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The presence of various proteins, including modified ones, in food which exhibit diverse immunogenic and sensitizing properties increases the difficulty of predicting host immune responses. Still, there is a lack of sufficiently reliable and comparable data and research models describing allergens in dietary matrices. The aim of the study was to estimate the immunomodulatory effects of β-lactoglobulin (β-lg) in comparison to those elicited by κ-casein (κ-CN), in vivo and ex vivo, using naïve splenocytes and a mouse sensitization model. Our results revealed that the humoral and cellular responses triggered by β-lg and κ-CN were of diverse magnitudes and showed different dynamics in the induction of control mechanisms. β-Lg turned out to be more immunogenic and induced a more dominant Th1 response than κ-CN, which triggered a significantly higher IgE response. For both proteins, CD4 +  lymphocyte profiles correlated with CD4 + CD25 +  and CD4 + CD25 + Foxp3 +  T cells induction and interleukin 10 secretion, but β-lg induced more CD4 + CD25 + Foxp3 -  Tregs. Moreover, ex vivo studies showed the risk of interaction of immune responses to different milk proteins, which may exacerbate allergy, especially the one caused by β-lg. In conclusion, the applied model of in vivo and ex vivo exposure to β-lg and κ-CN showed significant differences in immunoreactivity of the tested proteins (κ-CN demonstrated stronger allergenic potential than β-lg), and may be useful for the estimation of allergenic potential of various food proteins, including those modified in technological processes.

Differences in Regulatory Mechanisms Induced by β-Lactoglobulin and κ-Casein in Cow’s Milk Allergy Mouse Model–In Vivo and Ex Vivo Studies