[调节性T细胞及相关细胞因子在小鼠同种异体角膜移植排斥反应中的表达]
[Expression of regulatory T cells and related cytokines in mice allogenic corneal transplantation rejection].
作者信息
Wang Xin, LE Qi-Hua, Hong Jia-Xu, Xu Jian-Jiang, Sun Xing-Huai, Wang Wen-Tao, Zhuang Hong
机构信息
Department of Ophthalmology, Eye, Ear, Nose and Throat Hospital of Fudan University, Shanghai 200031, China.
出版信息
Zhonghua Yan Ke Za Zhi. 2009 Dec;45(12):1118-26.
OBJECTIVE
To study the changes of CD4(+)CD25(+) and CD8(+)CD103(+) regulatory T cells and their relevant cytokines in corneal allograft transplantation in mice.
METHODS
BABL/c (H-2(d)) mice received corneal allografts from C3H/He (H-2(k)) mice were served as the experimental group. BABL/c (H-2(d)) mice received corneal from BABL/c (H-2(d)) mice were used as the control group. Corneal graft survival time was recorded pre-, 3rd day, 7th day, 4th week, 8th week post-operatively. Infiltration of inflammatory cells and corneal neovascularization were evaluated by histopathologic examination. The percentage of CD4(+)CD25(+) and CD8(+)CD103(+) T cell in peripheral blood and spleen was determined by flow cytometry. The expression of interleukin-10 (IL-10), IL-4, gamma-interferon (IFN-gamma) and IL-1beta in serum and aqueous humor was measured by enzyme-linked immunospecific assay (ELISA).
RESULTS
The graft rejection in experimental group occurred at 7 days to 4 weeks after the operation, averaged (14.79 +/- 1.02) days. The grafts in the control group remained clear within 8 weeks observation and the survival time is much longer than that of the allografts (chi(2) = 46.934, P = 0.000). Flow cytometry showed that the percentage of CD4(+)CD25(+) T cell in peripheral blood in the control group after surgery was (3.36 +/- 0.29)%, (4.09 +/- 0.44)%, (5.44 +/- 0.35)%, (5.73 +/- 0.53)% at day 3, day 7, 4th week, and 8th week, respectively, which was significantly higher than that in the experimental group [(2.50 +/- 0.39)%, (3.24 +/- 0.25)%, (4.20 +/- 0.45)%, (4.18 +/- 0.14)%; t = 3.828, 2.898, 3.780, 4.892; P < 0.05]. On the other hand, CD8(+)CD103(+) T cell in peripheral blood in the experimental group after surgery was (2.20 +/- 0.33)%, (2.79 +/- 0.57)%, (4.55 +/- 1.03)%, (4.31 +/- 0.07)% at different periods, which was much higher than that in the control group (t = 7.133, 4.876, 5.196, 19.960; P < 0.05). The changes of CD4(+)CD25(+) and CD8(+)CD103(+) T cells in the spleen was earlier than those in peripheral blood. ELISA showed the expression of IL-10 and IL-4 in the serum in experimental group after surgery was much lower than that in the control group (t = 3.203, 3.141, 3.012, 2.869 and 2.340, 6.681, 8.839, 8.574; P = 0.011, 0.012, 0.013, 0.019 and 0.053, 0.000, 0.000, 0.000). The serum levels of IFN-gamma and IL-1beta in experimental group were higher than that in the control group (t = 3.508, 3.265, 4.402, 5.539 and 3.630, 5.796, 1.728, 0.660; P = 0.006, 0.011, 0.002, 0.000 and 0.005, 0.000, 0.115, 0.524). Furthermore, the cytokine levels in the aqueous humor behaved similarly with those in the serum.
CONCLUSION
The down-regulation of CD4(+)CD25(+) Treg, IL-10 and IL-4 and the up-regulation of CD8(+)CD103(+) Treg, IFN-gamma and IL-1beta in mice allograft corneal rejection may play an important role in allograft rejection.
目的
研究小鼠角膜移植中CD4(+)CD25(+)和CD8(+)CD103(+)调节性T细胞及其相关细胞因子的变化。
方法
将接受C3H/He（H-2(k)）小鼠角膜移植的BABL/c（H-2(d)）小鼠作为实验组。将接受BABL/c（H-2(d)）小鼠角膜的BABL/c（H-2(d)）小鼠作为对照组。记录术前、术后第3天、第7天、第4周、第8周角膜移植的存活时间。通过组织病理学检查评估炎性细胞浸润和角膜新生血管形成。采用流式细胞术测定外周血和脾脏中CD4(+)CD25(+)和CD8(+)CD103(+) T细胞的百分比。采用酶联免疫吸附测定（ELISA）法检测血清和房水中白细胞介素-10（IL-10）、IL-4、γ-干扰素（IFN-γ）和IL-1β的表达。
结果
实验组移植排斥反应发生在术后7天至4周，平均（14.79±1.02）天。对照组移植片在8周观察期内保持透明，存活时间远长于同种异体移植片（χ(2)=46.934，P=0.000）。流式细胞术显示，术后对照组外周血中CD4(+)CD25(+) T细胞百分比在第３天、第7天、第4周和第8周分别为（3.36±0.29）%、（4.09±0.44）%、（5.44±0.35）%、（5.73±0.53）%，显著高于实验组[（2.50±0.39）%、（3.24±0.25）%、（4.20±0.45）%、（4.18±0.14）%；t=3.828、2.898、3.780、4.892；P<0.05]。另一方面，术后实验组外周血中不同时期CD8(+)CD103(+) T细胞百分比分别为（2.20±0.33）%、（2.79±0.57）%、（4.55±1.03）%、（4.31±0.07）%，远高于对照组（t=7.133、4.876、5.196、19.960；P<0.05）。脾脏中CD4(+)CD25(+)和CD8(+)CD103(+) T细胞的变化早于外周血。ELISA显示，术后实验组血清中IL-10和IL-4的表达远低于对照组（t=3.203、3.141、3.012、2.869 和 2.340、6.681、8.839、8.574；P=0.011、0.012、0.013、0.019 和 0.053、0.000、0.000、0.000）。实验组血清中IFN-γ和IL-1β水平高于对照组（t=3.508、3.265、4.402、5.539 和 3.630、5.796、1.728、0.660；P=0.006、0.011、0.002、0.000 和 0.005、0.000、0.115、0.524）。此外，房水中细胞因子水平与血清中表现相似。
结论
小鼠同种异体角膜移植排斥反应中CD4(+)CD25(+)调节性T细胞、IL-10和IL-4的下调以及CD8(+)CD103(+)调节性T细胞、IFN-γ和IL-1β的上调可能在同种异体移植排斥反应中起重要作用。
Zotero 插件