细胞因子和树突状细胞参与超抗原SEB诱导高危角膜移植免疫抑制的研究

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目的探讨细胞因子和树突状细胞(DCs)在细菌性超抗原金黄色葡萄球菌外毒素B亚单位(SEB)诱导的高危角膜移植免疫抑制中的作用,比较SEB与糖皮质激素作用的差异。设计实验性研究。研究对象Fisher 344和Lewis纯系大鼠。方法供受体分别为纯系大鼠Fisher344和Lewis,SEB组和对照组分别于术前腹腔注射0.2ml SEB(75μg/ml)和生理盐水0.2ml/1次/4日,共3次;地塞米松组术后第1天开始结膜下注射地塞米松0.1ml(1mg/ml),1次/日,共2周。缝线法诱导受体角膜新生血管,穿透性角膜移植术后观察植片的存活状况,液相芯片技术检测术前及术后第10、30天房水和外周血中细胞因子含量,免疫组织化学方法检测角膜植片细胞因子及DCs表达情况。主要指标角膜植片平均存活时间,房水、外周血、角膜植片中细胞因子含量。结果与对照组角膜植片平均存活时间(MST)比较,SEB组的MST延长3.8天(P=0.00),地塞米松组MST延长7.1天(P= 0.01),SEB组和地塞米松组MST差异无统计学意义(P=0.26)。液相芯片检测显示,SEB组房水中IL-1β含量较对照组减少,IFN-γ、IL-4,IL-6含量明显升高,外周血仅可检出IFN-γ、IL-6两种细胞因子,变化趋势同房水中改变。免疫组化显示SEB组植片中IL-2较对照组明显减少,而IFN-γ、IL-4、IL-6明显升高,植片中DCs表达接近对照组,但表型不同。对照组以成熟DCs为主,SEB组以未成熟DCs为主。结论SEB在防治高危角膜移植免疫排斥反应方面与糖皮质激素无差异,但作用机制不同。SEB通过影响眼部细胞因子的产生调节免疫反应,诱导免疫抑制,DCs也可能参与作用。 Objective To investigate the role of cytokines and dendritic cells (DCs) in the immunosuppression of high-risk corneal transplantation induced by bacterial superantigen Staphylococcus aureus exotoxin B subunit (SEB), and to compare the effects of SEB with glucocorticoid. Design experimental research. Study subjects Fisher 344 and Lewis pure-line rats. Methods The recipients were fed with Fisher344 and Lewis, respectively. The SEB group and the control group were injected intraperitoneally with 0.2ml SEB (75μg / ml) and saline 0.2ml / 1 times / On the first day after the operation, dexamethasone 0.1ml (1mg / ml) was given subconjunctivally in the dexamethasone group once a day for 2 weeks. The suture was used to induce the corneal neovascularization. After penetrating keratoplasty, the survival of the grafts was observed. The contents of cytokines in the aqueous humor and peripheral blood were measured before and after the 10th and 30th days Histochemical methods were used to detect the expression of cytokines and DCs in corneal graft. The main indicators of corneal graft survival time, aqueous humor, peripheral blood, corneal graft cytokines content. Results Compared with the control group, the mean survival time (MST) of corneal graft in SEB group was 3.8 days (P = 0.00), MST in dexamethasone group was 7.1 days (P = 0.01), MST in SEB group and dexamethasone group The difference was not statistically significant (P = 0.26). The liquid chip detection showed that the content of IL-1β in aqueous humor of SEB group was lower than that of the control group, the content of IFN-γ, IL-4 and IL-6 were significantly increased, and only IFN-γ and IL- Cytokines, changes in the same room with changes in water. Immunohistochemistry showed that the level of IL-2 in the SEB group was significantly lower than that in the control group, while the levels of IFN-γ, IL-4 and IL-6 in the SEB group were significantly higher than those in the control group. The control group is dominated by mature DCs, while the SEB group is dominated by immature DCs. Conclusion SEB has no difference with glucocorticoid in the prevention and treatment of high-risk keratoplasty rejection, but its mechanism of action is different. SEB can regulate the immune response by affecting the production of cytokines in the eye and induce immunosuppression. DCs may also participate in the process.
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