INDUCTION AND REGULATION OF IMMUNE AND ALLERGIC RESPONSES BY FOOD-DERIVED IMMUNE FUNCTIONAL MOLECULES

1. 肠道免疫特征的分子生物学分析(1)DNA微阵列分析鉴定出IEL中比脾脏T细胞表达更丰富的基因。(2)我们还阐明了卵清蛋白（OVA）特异性t细胞受体转基因（TCR-Tg）小鼠饲喂蛋清饮食后，EEL中基因表达模式的变化。(3)建立了小鼠胎肠上皮细胞原代培养方法，并验证了其抗原提呈能力。口服耐受诱导机制分析(1)我们发现ppp缺乏小鼠可诱导口服耐受。(2)采用双向电泳和质谱分析方法测定口服耐受CD4 T细胞特异性表达的细胞内蛋白。结果表明，Caspase 3在口服耐受CD4 T细胞中上调。(3)利用减法杂交技术克隆了口服耐受CD4 T细胞中特异性表达的基因。食物过敏模型分析及食物源性免疫功能分子对食物过敏反应的抑制作用。(1) OVA特异性TCR-Tg小鼠饲喂OVA日粮，不仅可诱导血清IgE反应，还可引起小肠炎症反应。饲喂OVA诱导的OVA特异性抗体主要识别OVA的三级结构。(2)在这种食物过敏模型中，OVA特异性IgE的产生和体重的下降被证明可以通过给予抗原OVA肽的tcr拮抗类似物来抑制。(3)在该食物过敏模型中，饲喂棉子糖抑制了IgE反应，棉子糖具有调节肠道菌群的作用。(4)我们还发现，初始CD4+ T细胞的细胞因子分泌模式随着ova衍生肽作为抗原肽的剂量而改变。

1. Molecular biological analysis of characteristics of gut immunity (1) DNA microarray analysis identified genes expressed more abundantly in IEL than in spleen T cells. (2) We also elucidated the change in the gene expression pattern in EEL when ovalbumin (OVA)-specific T-cell receptor transgenic (TCR-Tg) mice were fed an egg-white diet. (3) The method for the primary culture of murine fetal intestinal epithelial cells was established and its capability for antigen presentation was shown.2. Analysis of mechanisms of oral tolerance induction (1) We showed that oral tolerance was induced in PP-deficient mice. (2) Intracellular proteins specifically expressed in orally tolerized CD4 T cells were determined by two-dimensional electrophoresis and mass spectrometric analysis. The results demonstrated that Caspase 3 was upregulated in orally tolerant CD4 T cells. (3) Genes expressed specifically in orally tolerized CD4 T cells were cloned using subtractive hybridization.3. Analysis of food allergy model, and suppression of food allergic reactions by food-derived immune functional molecules. (1) Feeding OVA diet to OVA-specific TCR-Tg mice resulted not only in induction of a serum IgE response, but also in inflammation in the small intestine. OVA-specific antibodies elicited by feeding OVA recognized mainly the tertiary structure of OVA. (2) OVA-specific IgE production and the decrease in body weight induced in this food-allergy model were shown to be suppressed by administering a TCR-antagonistic analog of the antigenic OVA peptide. (3) Feeding raffinose, which is known to modulate intestinal microflora, suppressed the IgE response in this food allergy model. (4) We also showed that the cytokine secretion pattern of naive CD4+ T cells were altered depending on the dose of OVA-derived peptide as an antigenic peptide.