FTY720和环孢素A局部应用可延长小鼠角膜移植的存活时间。
Topical application of FTY720 and cyclosporin A prolong corneal graft survival in mice.
作者信息
Liu Yong, Jiang Jingjing, Xiao He, Wang Xiaokui, Li Yan, Gong Yubo, Wang Dajiang, Huang Yifei
机构信息
Departments of Ophthalmology, Chinese PLA General Hospital, Beijing, China.
出版信息
Mol Vis. 2012;18:624-33. Epub 2012 Mar 9.
PURPOSE
To investigate the effects of topical FTY720 and cyclosporin A (CsA) on allogeneic corneal transplantation in mice.
METHODS
A total of 75 BALB/c mice received corneal grafts from C57BL/6 donors. Recipients were treated with 0.1%, 0.3%, or 0.5% FTY720 ophthalmic gel or 1% CsA eye-drops after the graft (controls received no treatment). The number of cluster of differentiation (CD)4+ T cells and CD4+CD25+forkhead box P3 (Foxp3)+ regulatory (Treg) cell phenotypes were measured by flow cytometry. Cytokine mRNA expression in corneal grafts was analyzed by real-time quantitative PCR. CD4 + T cells and cytokines in corneal samples were identified by immunohistochemical staining.
RESULTS
Corneal graft survival was prolonged by treatment with topical 0.5% FTY720 (mean survival time [MST], 24.1±1.6 days) or 1% CsA eye-drops (MST 25.0±1.9 days) compared with controls (MST, 13.4±0.5 days; n=9, both p<0.01). Topical 0.5% FTY720 treatment significantly increased the percentages of CD4 + T (p<0.05) and Treg cells (p<0.01; n=5) in the cervical lymph nodes compared with controls. Transforming growth factor-β1 (TGF-β1) mRNA transcription in corneal grafts after topical 0.5% FTY720 increased (p<0.05, n=3), while interleukin-2 (IL-2) and interferon-γ (IFN-γ) mRNA expression in corneal grafts treated with 1% CsA decreased (p<0.01, p<0.05, respectively). These cytokine results were paralleled by similar immunohistochemical staining. Topical 0.5% FTY720 and 1% CsA treatment reduced the infiltration of CD4+ Tcells in the grafts.
CONCLUSIONS
Topical 0.5% FTY720 and 1% CsA can effectively prolong allogeneic corneal graft survival in mice. Treatment with topical 0.5% FTY720 increases the percentage of CD4+ T cells and the percentage of Treg cells in cervical lymph nodes. The 0.5% FTY720 increased TGF-β1 mRNA expression and decreases infiltration of CD4+ T cells in corneal grafts, while topical 1% CsA down-regulated the expression of IL-2 and IFN-γ.
目的
研究局部应用FTY720和环孢素A（CsA）对小鼠同种异体角膜移植的影响。
方法
75只BALB/c小鼠接受来自C57BL/6供体的角膜移植。移植后，受体分别用0.1%、0.3%或0.5%的FTY720眼凝胶或1%的CsA滴眼液治疗（对照组不治疗）。通过流式细胞术检测分化簇（CD）4⁺T细胞和CD4⁺CD25⁺叉头框P3（Foxp3）⁺调节性（Treg）细胞表型的数量。通过实时定量PCR分析角膜移植中细胞因子mRNA的表达。通过免疫组织化学染色鉴定角膜样本中的CD4⁺T细胞和细胞因子。
结果
与对照组（平均生存时间[MST]，13.4±0.5天；n = 9，p均<0.01）相比，局部应用0.5%的FTY720（MST，24.1±1.6天）或1%的CsA滴眼液（MST 25.0±1.9天）可延长角膜移植的存活时间。与对照组相比，局部应用0.5%的FTY720治疗可显著增加颈部淋巴结中CD4⁺T细胞（p<0.05）和Treg细胞（p<0.01；n = 5）的百分比。局部应用0.5%的FTY720后，角膜移植中转化生长因子-β1（TGF-β1）mRNA转录增加（p<0.05，n = 3），而用1%的CsA治疗的角膜移植中白细胞介素-2（IL-2）和干扰素-γ（IFN-γ）mRNA表达降低（分别为p<0.01，p<0.05）。这些细胞因子结果与类似的免疫组织化学染色结果一致。局部应用0.5%的FTY720和1%的CsA治疗可减少移植中CD4⁺T细胞的浸润。
结论
局部应用0.5%的FTY720和1%的CsA可有效延长小鼠同种异体角膜移植的存活时间。局部应用0.5%的FTY720可增加颈部淋巴结中CD4⁺T细胞百分比和Treg细胞百分比。0.5%的FTY720可增加角膜移植中TGF-β1 mRNA表达并减少CD4⁺T细胞浸润，而局部应用1%的CsA可下调IL-2和IFN-γ的表达。
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