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  

Study on cytokines and dendritic cells involving immunologic inhibition of rat high-risk corneal transplantation induced by SEB

作　　者：谭小波[1] 潘志强[1] 陈钰[2] 接英[1] 张文华[1] 武宇影[1] 

机构地区：[1]首都医科大学附属北京同仁医院眼科中心 北京市眼科研究所,100730 [2]解放军总医院免疫学教研室

出　　处：《眼科》2007年第3期169-174,共6页Ophthalmology in China

基　　金：国家自然科学基金(30271385);教育部新世纪优秀人才支持计划资助

摘　　要：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 explore the role of cytokines and dendritic cells （DCs） in rat high-risk corneal allograft survival prolonged by superantigen Staphylococcal Enterotoxin B （SEB） and to compare the different effects between SEB and glucocorticoid. Design Experimental study. Participants Fisher 344 and Lewis rats. Methods The Fisher 344 and Lewis inbred rats were used for high-risk penetrating keratoplasty model. All of the Lewis rat recipients were divided into three groups by blinded fashion. Group Ⅰ and Group Ⅱ rats were injected intraperitoneally with 0.2ml saline buffer or SEB （75μg/ml） respectively at 4-day intervals on three occasions before transplantation. Group Ⅲ rats were injected subeonjunctivally with 0.1ml dexamethasone （1mg/ml） daily from the first day after surgery for 2 weeks. The allograft survival was examined under slit-lamp. The concentration of interleukin IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, TNF-α in rat aqueous humor and peripheral blood were measured by liquichip and the cytokine and CD11c, CD80, MHC-Ⅱ expression in corneal grafts were examined by immunohistochemestry staining. Main Octcome Measures Mean survival time of the allografts, the level of cytokines in aqueous humor, peripheral blood and corneal grafts. Results Compared with group control, the grafts mean survival time was delayed about 3.8d in group SEB （P=0.00） and 7.1d in group dexamethasone（P=0.01）. But there was no significant difference between group SEB and dexamethone （P=0.26）. Liquichip test showed that the level of IL-1β in aqueous humor was reduced and IL-4, IL-6 and IFN-γ ascended. Only IFN-γ and IL-6 could be found in peripheral blood, and the changing shift of them was similar to that in aqueous humor. The immunohistochemistry staining showed that the expression of IL-2 in rat corneal grafts was signif- icantly decreased but IL-4, IL-6 and IL-10 elevated. The expression of DCs in group SEB was similar to that in group control, which was elevated after keratoplasty, but the pheno

关 键 词：超抗原 角膜移植 细胞因子 树突状细胞 免疫耐受 

分 类 号：R779.65[医药卫生—眼科]