Glycation is a spontaneous reaction accompanying the thermal processing and storage of food. It can lead to changes in the allergenic and immunogenic potential of protein. This study aimed to evaluate the effect of the glycation of α-lactalbumin and β-lactoglobulin (β-lg) on the ex vivo response of β-lg sensitized lymphocytes. C57BL/6 mice were immunized intragastrically (i-g) or intraperitoneally (i-p) with β-lg. The humoral response of the groups differed only with respect to the IgE level of the i-p group. Cellular response was studied after stimulation with antigen variants. The lymphocytes from the i-g/group mesenteric lymph nodes, stimulated with β-lg before and after glycation, presented a higher percentage of CD4 and CD8 T cells compared to the i-p/group. The cytokine profile of the i-p/group splenocytes stimulated with antigens showed elevated levels of pro-inflammatory IL-17A regardless of protein modification. In conclusion, the ex vivo model proved that the glycation process does not reduce protein immunogenicity.