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目的观察完整无损伤正常皮肤外用抗原诱导免疫耐受的效应,并探讨其可能的机制。方法 Balb/c小鼠背部皮肤外用不同浓度卵清蛋白(ovalbumin,OVA),1次/周,连用5周。第6周始予皮下注射OVA,1次/周,共3周。检测血清标本OVA特异性抗体水平。体外培养脾脏细胞,给予OVA刺激,分析T细胞分泌细胞因子情况。结果皮肤外用OVA抗原5周后,小鼠血清中未检测到抗OVA抗体;皮下OVA免疫3次后,对照小鼠血清有高滴度的抗OVAIgG1,IgG2a和IgE抗体,而1000μgOVA抗原皮肤外用组的小鼠血清OVA特异性IgG1和IgE抗体水平显著低于其他剂量的外用组(P<0.05),其脾脏细胞培养上清中IL-2,IL-4,IL-6和IFN-γ细胞因子水平也显著低于0μg,10μg和100μg抗原外用组。结论正常无损伤皮肤外用抗原可以产生免疫耐受效应,高浓度抗原皮肤外用可以有效抑制皮下免疫所诱导的免疫应答。
Objective To observe the effect of intact normal skin antigen on immune tolerance and to explore its possible mechanism. Methods Balb / c mice were orally treated with ovalbumin (OVA) once a week for 5 weeks. Subcutaneous injection of OVA from the 6th week was given once a week for 3 weeks. Serum samples were tested for OVA-specific antibody levels. Spleen cells were cultured in vitro and stimulated with OVA to analyze the cytokine secretion by T cells. Results After OVA antigens were applied to the skin for 5 weeks, no anti-OVA antibodies were detected in the serum of the mice. Anti-OVA IgG1, IgG2a and IgE antibodies were highly titrated in the serum of the control mice after three times immunization with the subcutaneous OVA, while 1000 μg OVA antigen (P <0.05). The levels of IL-2, IL-4, IL-6 and IFN-γ cytokines in the spleen cell culture supernatant of OVA-specific IgG1 and IgE were significantly lower than those of other groups Levels were also significantly lower than the 0 μg, 10 μg and 100 μg antigens topical groups. Conclusion The normal non-destructive skin external antigens can produce immune tolerance effect, and the high concentration anti-skin external application can effectively inhibit the immune response induced by subcutaneous immune.