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目的研究婴儿型双歧杆菌对花生过敏小鼠肠道Th2型反应的调节作用。方法通过应用花生蛋白诱导肠道的Th2型反应,建立食物过敏小鼠模型。过敏小鼠灌胃给予婴儿型双歧杆菌(ATCC菌或CGMCC0313-2)或不做处理。然后分离小鼠小肠黏膜CD4+T细胞或DC,另取肠黏膜组织进行石蜡包埋甲苯胺蓝染色肥大细胞计数,HE染色进行嗜酸细胞和单个核细胞计数,流式细胞检测CD4+T中Th2(CD4+IL-4+T)细胞和Treg(CD4+CD25+Foxp3+T)比例,另取CD4+T进行CFSE标记,与DC共培养4 d后流式细胞检测CD4+T增殖反应,收集细胞培养液ELISA检测IL-4、IL-5和IL-13分泌水平。结果过敏组小鼠Th2型细胞数,CD4+T细胞增殖反应,IL-4、IL-5和IL-13水平,肠黏膜中肥大细胞、嗜酸性细胞和单个核细胞数均明显高于对照组(P<0.01),而Treg数目低于对照组(P<0.01),婴儿双歧杆菌干预后,婴儿双歧杆菌组Th2型细胞数,IL-4、IL-5和IL-13水平,肠黏膜中肥大细胞、嗜酸性细胞和单个核细胞数均明显低于过敏组(P<0.01),而Treg数目高于过敏组(P<0.01)。结论口服婴儿型双歧杆菌可以抑制花生过敏导致的肠道Th2型反应。
Objective To study the regulatory effect of Bifidobacterium infantis on intestinal Th2 response in peanut allergy mice. Methods By using peanut protein to induce the Th2 response in the intestine, a mouse model of food allergy was established. Allergic mice were given intranasal Bifidobacterium infantis (ATCC bacteria or CGMCC0313-2) or untreated. CD4 + T cells or DCs from mouse intestinal mucosa were isolated, and other intestinal mucosa tissues were paraffin-embedded for toluidine blue staining for mast cell counts. Eosinophils and mononuclear cells were counted with HE staining. Flow cytometry was used to detect CD4 + T cells Th2 CD4 + T CD4 + T cells and Treg CD4 + T CFSE markers were co-cultured with DC 4 days after flow cytometry CD4 + T proliferative response, Cell culture medium ELISA was used to detect IL-4, IL-5 and IL-13 secretion levels. Results The numbers of Th2 type cells, the proliferation of CD4 + T cells, the levels of IL-4, IL-5 and IL-13, the numbers of mast cells, eosinophils and mononuclear cells in the allergic mice were significantly higher than those in the control group (P <0.01), and the number of Tregs was lower than that of the control group (P <0.01). After Bifidobacterium infantis intervention, the levels of Th2 cells, IL-4, IL-5 and IL-13 in Bifidobacterium infantis group, Mucous mucosa, eosinophils and mononuclear cells were significantly lower than the allergic group (P <0.01), while the number of Treg was higher than the allergic group (P <0.01). Conclusions Oral infant bifidobacteria can inhibit the intestinal Th2 response induced by peanut allergy.