Exogenous IL-10 induces corneal transplantation immune tolerance by a mechanism associated with the altered Th1/Th2 cytokine ratio and the increased expression of TGF-β
Exogenous IL-10 induces corneal transplantation immune tolerance by a mechanism associated with the altered Th1/Th2 cytokine ratio and the increased expression of TGF-β
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DOI：
10.3892/mmr.2014.2073
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发表时间：
2014-06-01
影响因子：
3.4
通讯作者：
Wang, Xuefeng
中科院分区：
文献类型：
--
作者：
Li, Bing;Tian, Lihua;Wang, Xuefeng
本研究旨在探讨外源性IL-10基因修饰的大鼠树突状细胞(DCs)在同种异体角膜移植中的作用。采用大鼠淋巴细胞分离液和细胞因子诱导法提取和培养大鼠骨髓来源的树突状细胞前体细胞。以SD大鼠为受体，Wistar大鼠为供体，建立角膜移植模型。流式细胞仪检测成熟树突状细胞CD83和细胞表面特异性刺激分子CD86的表达。用Western印迹法检测IL-10蛋白表达，RT-PCR法检测细胞因子IL-10、IL-2和转化生长因子-βmRNA的表达。与其他组相比，IL-10绿色荧光蛋白(GFP)-DC组角膜移植物存活时间明显延长，HE染色显示角膜移植物轻度水肿和炎性细胞浸润。IL-10-GFP-DC组高表达IL-10，低表达表面抗原CD83和CD86，T淋巴细胞增殖能力低。IL-10-GFP-DC组IL-10和TGF-β的表达高于其他组，而IL-2的表达低于其他组。IL-10基因的导入抑制了树突状细胞的成熟。IL-10基因修饰的未成熟树突状细胞(IMDC)能够抑制同种异体角膜移植排斥反应，诱导免疫耐受，延长移植角膜的存活时间。Th1/Th2偏离和转化生长因子-β的高表达可能导致免疫耐受。
The aim of the present study was to examine the effect of exogenous IL-10 transfected rat dendritic cells (DCs) in corneal allografts. Rat lymphocyte separation medium and a cytokine induction method was used to extract and culture precursor cells of rat bone marrow-derived dendritic cells. A corneal transplant model was established, with Sprague-Dawley (SD) rats as the recipients and Wistar rats as the donors. Flow cytometry (FCM) was used to detect the expression of CD83, which indicates a mature dendritic cell, and a specific cell surface stimulatory molecule CD86. Western blot analysis was used to detect interleukin (IL)-10 protein expression and RT-PCR was used to detect cytokine IL-10, IL-2 and TGF-beta mRNA expression in each group. Compared with the other groups, the survival time of corneal grafts in the IL-10-green fluorescent protein (GFP)-DC group was significantly prolonged and H&E staining demonstrated mild graft edema and inflammatory cell infiltration. There was a high expression of IL-10 and a low expression of the surface antigens, CD83 and CD86, with a lower proliferation of T lymphocytes in the IL-10-GFP-DC group. The expression of IL-10 and TGF-beta in the IL-10-GFP-DC group was higher than that in the other groups, while the expression of IL-2 was lower. The transfection of the IL-10 gene inhibited dendritic cell maturation. IL-10 gene-modified immature dendritic cells (imDC) were able to inhibit corneal allograft rejection and induce immune tolerance to prolong the survival time of the corneal graft. The Th1/Th2 deviation and the high expression of TGF-beta may lead to immune tolerance.