细胞毒T淋巴细胞相关抗原4免疫球蛋白抑制鼠高危角膜移植免疫排斥反应的实验研究  

CTLA4-Ig prevents corneal allograft reject ion in mice

作　　者：史伟云[1] 谢立信[1] 

机构地区：[1]山东省眼科研究所暨眼科医院,青岛266071

出　　处：《中华眼科杂志》2004年第10期696-700,共5页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金资助项目 ( 3 0 2 712 3 9);山东省自然科学基金资助项目 (Y2 0 0 2C14 )

摘　　要：目的　探讨细胞毒T淋巴细胞相关抗原 4免疫球蛋白 (CTLA4 Ig)对小鼠高危角膜移植免疫排斥反应的抑制作用及局部抗免疫排斥反应机制。方法　建立 5 0只BALB/c小鼠穿透性角膜移植动物模型。治疗组 (2 5只 ) :取C5 7BL/ 6小鼠角膜片 ,放置于 10 μg/mlCTLA4 Ig保存液中浸泡 2 4h后 ,移植到BALB/c小鼠。对照组 (2 5只 ) :植片不行任何处理。术后每 3d用裂隙灯显微镜检查植片情况 ,每周应用组织学和免疫组织化学方法检测植片中各种炎性细胞和淋巴细胞的变化。对出现排斥反应的角膜植片应用逆转录PCR(RT PCR)方法检测部分细胞因子的表达。另外 ,选择经CTLA4 Ig治疗、植片保持透明 6周以上的小鼠作为受体 ,接受来自C5 7BL/ 6小鼠皮肤的移植 ,当移植皮肤发生排斥时 ,进行迟发性超敏反应 (DTH)分析。结果　CTLA4 Ig治疗组角膜植片保持透明 >10 0d。对照组小鼠术后 14d内均发生免疫排斥反应。组织病理学检查显示 ,角膜移植术后 2周 ,CTLA4 Ig治疗组植片保持正常细胞结构 ,无明显炎性细胞和T淋巴细胞浸润 ;对照组的排斥植片有大量炎性细胞和T淋巴细胞浸润 (包括CD+ 4 ,CD+ 8及CD+ 11细胞 )。术后 2周发生免疫排斥的角膜植片中检测到白细胞介素 10 (IL 10 ) ,肿瘤坏死因子α(TNF α) ,γ干扰素 (IFN γ) 。Objective Selective inhibition of T cell activation using the B7 specific fusio n protein CTLA4-Ig has been shown to block CD28/B7 signal pathway. In this s tudy, the capacity of soluble CTLA4-Ig alone in the suppressing corneal allogra ft rejection is further tested in mice, and its mechanism of preventing the corn ea from immune rejection in local eye is analyzed.Methods The mouse models of corneal allografts were established. In one group(25 mice), the corneal donors from C57BL/6 mice were incubated in corneal storage medium c ontaining 10 μg/ml of CTLA4-Ig for 24 hours, and then transplanted orthotopica lly into the recipients cornea of BALB/c mice. In another group(25 mice), as a control, no treatment was conducted on the donor corneas before surgery. The co ndition of the allografts was monitored using slit lamp microscopy every 3 days and the cellar architecture of selected graft was examined by histological and i mmunohistochemical techniques weekly. Local corneal cytokines expressing in the rejected grafts were examined. Some mice, which were treated with CTLA4-Ig a nd their grafts survived beyond 6 weeks, were selected as recipients of skin gra fts from C57BL/6 donors. Delayed type hypersensitivity (DTH) was measured afte r the skin grafts rejection.Results All mice in the control group were rejected within 14 days after transplantation , while grafts incubated in CTLA4-Ig for 24 hours survived well beyond 100 days . The histopathology of surviving allografts had a normal cellular architecture , whereas the rejected allografts were heavily infiltrated with inflammatory cel ls and T lymphocytes ( including CD+ 4, CD+ 8, and CD+ 11 cells) . The cytokines including IL-10, TNF-α, IFN-γ, B7-1 and CD40 were detecte d in the rejected corneas. Skin grafts from the C57BL/6 donor into the BALB/c mice after corneal transplantation were rejected with the expected median survi val time of 11 days. DTH ear swelling challenge was induced after skin rejectio n, and a significant difference wa

关 键 词：免疫排斥反应 术后 植片 CTLA4-IG 角膜移植 高危 相关抗原 结论 保持 情况 

分 类 号：R779.65[医药卫生—眼科] R392[医药卫生—临床医学]