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目的观察和比较致敏小鼠及正常小鼠DC表面共刺激分子表达及CD4+CD25+Foxp3+T数量的差异及大剂量过敏原在体外的作用。方法流式细胞仪检测致敏及正常对照小鼠脾脏DC表面分子CD11c、MHCⅡ、CD80、CD86表达。分离致敏及正常对照小鼠CD4+T细胞,流式细胞仪检测CD4+CD25+Foxp3+T细胞的数量。致敏小鼠脾脏DC、CD4+T细胞与10 mg/ml OVA或生理盐水共培养后,流式细胞仪检测并比较CD80、CD86等共刺激分子的表达及CD4+CD25+Foxp3+T细胞的数量。结果致敏小鼠脾脏DC表面共刺激分子CD80、CD86、MHCⅡ表达显著高于正常对照小鼠。10 mg/ml的OVA作用后,致敏小鼠脾脏DC表面共刺激分子CD80、CD86的表达明显降低。致敏小鼠脾脏细胞中CD4+CD25+Foxp3+T细胞数量显著低于正常对照小鼠。10 mg/ml的OVA作用后,致敏小鼠CD4+CD25+Foxp3+T细胞数量显著上升。结论大剂量过敏原在体外诱导致敏小鼠T细胞的不反应性,其机制与降低致敏小鼠DC共刺激分子表达,诱导调节性T细胞极化等有关。
Objective To observe and compare the expression of costimulatory molecules on DCs and the numbers of CD4 + CD25 + Foxp3 + T in sensitized mice and normal mice and the effect of high dose allergens in vitro. Methods Flow cytometry was used to detect the expression of CD11c, MHCⅡ, CD80 and CD86 in spleen of sensitized and normal control mice. CD4 + T cells from sensitized and normal control mice were isolated and the number of CD4 + CD25 + Foxp3 + T cells was detected by flow cytometry. CD4 + T cells of sensitized mice were co-cultured with 10 mg / ml OVA or normal saline. The expression of costimulatory molecules such as CD80 and CD86 were detected by flow cytometry and the expressions of CD4 + CD25 + Foxp3 + T cells Quantity. Results The expressions of costimulatory molecules CD80, CD86 and MHC Ⅱ on the surface of spleen DC in sensitized mice were significantly higher than those in normal control mice. After 10 mg / ml OVA treatment, the expression of co-stimulatory molecules CD80 and CD86 on the spleens DC in sensitized mice were significantly decreased. The number of CD4 + CD25 + Foxp3 + T cells in sensitized mice spleen cells was significantly lower than that in normal control mice. The number of CD4 + CD25 + Foxp3 + T cells in sensitized mice increased significantly after OVA treatment at 10 mg / ml. Conclusion The high dose of allergen induces the inactivation of T cells in sensitized mice in vitro. The mechanism is related to the reduction of DC costimulatory molecule expression and the induction of T cell polarization in sensitized mice.