摘要
目的研究高表达吲哚胺2,3-二氧化酶(in-doleamine 2,3-dioxygenase,IDO)的树突状细胞(dendritic cells,DC)对角膜植片存活时间的影响。方法采用脂质体转染的方法将质粒CTLA4Ig转入F344大鼠(供体)DC,用荧光显微镜观察目的基因的表达,采用RT-PCR方法检测转染前后DC中IDO mRNA的表达变化,应用流式细胞仪检测转染前后不同时间的DC对同种异体T细胞的影响。以F344大鼠为供体、Lewis大鼠为受体建立大鼠同种异体角膜移植模型,将基因修饰后高表达IDO的DC在术后立即注入Lewis大鼠体内(实验组),注射转染前DC的受体为对照组。用裂隙灯观察角膜植片的存活情况,术后第7天用八肽胆囊收缩素测定受体鼠T淋巴细胞对供体抗原的反应。结果转染CTLA4Ig促进了树突状细胞IDO的表达(P<0.05),并且转染后48h的DC能显著引起T细胞凋亡(P<0.05),注射高表达IDO的DC能明显延长角膜植片的存活时间(P<0.01),治疗组Lewis大鼠T淋巴细胞对供体的抗原刺激的反应明显低于对照组(P<0.05)。结论高表达IDO的供体DC能特异性抑制受体对供体抗原的免疫反应,明显延长角膜植片存活时间。
Objective To study the effects of dendritic cells （DCs） with high expression of indoleamine 2,3-dioxygenase （IDO） on corneal allografts.Methods Plasmid PG/CTLA4Ig was transfected into the dendritic cells of the F344 rat mediated by Lipofectamine^TM 2000. Expression of CTLA4Ig was detected by immuno-fluorescence. RT-PCR was used to detect the expression of IDOmRNA in dendritic cells. The flow cytometry assay in Annexin V/PI was used to detect the apoptosis of T cells after mixed lymphocyte cell reaction. The F344 rat was then used as the donor, and the Lewis rat was the receptor. Corneal transplants were performed. DCs with high expression of IDO were injected into Lewis rats on the day the transplants were performed and on the 3rd day after transplantation （treatment group）. A rejection reaction was observed using a slit lamp, and the survival of cemeal allografts was evaluated by Holland criterion. Mixed lymphocyte cell reaction of receptor to donor was performed by CCK8 on the 7th day after the operation.Results Expression of IDOmRNA increased in DEs after transfection （ P 〈 0.05）. DCs with high expression of IDO could induce the apoptosis of T cells. In the treatment group, the survival time for corneal allografts was significantly prolonged （P 〈 0. 01 ）, and the responsiveness of the receptor＇s T cells to the donor＇s antigens was significantly lower （ P 〈 0.05）. Conclusion Donor＇ s DCs with high expression of IDO could specifically suppress receptor＇ s immune response to donor＇ s antigen and significantly prolong the survival of corneal allografts.
出处
《眼视光学杂志》
CAS
2007年第2期90-93,共4页
Chinese Journal of Optometry & Ophthalmology
