The aim of this study was to evaluate the ex vivo and in vivo studies immune potential of α- and κ-casein. Ex vivo, naïve mouse splenocytes were stimulated with α- or κ-casein. After 120 h of culture, the proliferation index (PI), determined by 3-(4,5 dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and carboxyfluorescein diacetate N-succinimidyl ester (CFSE) staining, did not vary for either antigen, suggesting similar ex vivo immunogenic potential of both casein fractions. In vivo, BALB/ccmdb mice were sensitized with α- or κ-casein and then gavaged with primary antigen. Mice immunized with α-casein had higher levels of IgG (2^16.33) and IgA (2^10.22) in serum at the end of the experiment compared with mice immunized with κ-casein (2¹⁵ and 2^9.3 for IgG and IgA, respectively). The use of α-casein for mouse immunization and ex vivo lymphocyte stimulation resulted in higher concentrations of secreted cytokines (IL4, IL10) compared with κ-casein stimulation. This is consistent with increasing regulatory T cell (Treg) lymphocyte populations, independent of the antigen used for stimulation. In summary, the immunogenic potential of α- and κ-casein was similar. Humoral and cellular immune responses confirmed their strong, independent potential to induce B and T cells. We propose that the lymphocyte proliferation index be used as an initial screening for protein immunogenicity.

这项研究的目的是评估离体和体内研究α-酪蛋白和κ-酪蛋白的免疫潜能。用α-或κ-酪蛋白刺激离体的幼稚小鼠脾细胞。培养120小时后，通过3-（4,5二甲基噻唑-2基）-2,5-二苯基溴化四氮唑（MTT）测定和羧基荧光素二乙酸酯N-琥珀酰亚胺酯（CFSE）染色确定了增殖指数（PI）两种抗原均未改变，表明两种酪蛋白级分的相似的离体免疫原性潜力。在体内，将BALB / ccmdb小鼠用α-或κ-酪蛋白致敏，然后用第一抗原进行管饲。与用κ-酪蛋白免疫的小鼠（2 ）相比，在实验结束时用α-酪蛋白免疫的小鼠血清中的IgG（2 ^16.33）和IgA（2 ^10.22）水平更高。IgG和IgA分别为¹⁵和2 ^9.3）。与κ-酪蛋白刺激相比，使用α-酪蛋白进行小鼠免疫和离体淋巴细胞刺激可导致更高浓度的分泌细胞因子（IL4，IL10）。这与增加的调节性T细胞（Treg）淋巴细胞数量一致，而与用于刺激的抗原无关。总之，α-酪蛋白和κ-酪蛋白的免疫原性潜力相似。体液和细胞免疫反应证实它们具有强大的独立潜力来诱导B细胞和T细胞。我们建议将淋巴细胞增殖指数用作蛋白质免疫原性的初步筛选。