小鼠异基因角膜移植术后调节性T细胞与相关因子的表达研究  

Expression of regulatory T cells and related cytokines in mice aHogenic corneal transplantation rejection

作　　者：王欣[1] 乐琦骅[1] 洪佳旭[1] 徐建江[1] 孙兴怀[1] 王文韬[1] 庄宏[1] 

机构地区：[1]复旦大学附属眼耳鼻喉科医院眼科,上海200031

出　　处：《中华眼科杂志》2009年第12期1118-1126,共9页Chinese Journal of Ophthalmology

基　　金：基金项目：2007年卫生部临床学科重点项目;2008年复旦大学青年基金

摘　　要：目的探讨在小鼠异基因角膜移植免疫排斥中，调节性T细胞（Treg）及其下游细胞因子的表达变化。方法实验研究。实验组以BALB／c（H-2^d）小鼠为受体、C3H／He（H-2^k）小鼠为供体，建立小鼠异基因角膜移植模型48只；对照组的受体与供体均为BALB／c，建立小鼠同基因角膜移植模型48只。观察术后植片的存活率、排斥反应发生时间；组织病理学检查术后3d、7d、4周及8周各时间点角膜植片中炎症细胞浸润与角膜新生血管生成；流式细胞仪检测术前、术后3d、7d、4周及8周各时间点受体外周血和脾脏中CD4^＋CD25^＋Treg及CD103^＋CD8^＋Treg的表达变化；同时酶联免疫吸附试验检测血清与房水中自细胞介素10（IL-10）、IL-4、1干扰素（IFN—γ）及IL-1β的表达变化。应用Kaplan—Meier生存曲线、两独立样本t检验、非参数秩和检验进行统计学分析。结果实验组角膜植片免疫排斥发生时间为7d至4周，平均存活时间为（14．79±1．02）d，对照组术后植片保持透明，观察期（8周）内未发生排斥反应，存活时间显著长于实验组，差异有统计学意义（χ^2=46．934，P=0．000）。流式细胞仪检测显示对照组外周血CIN^＋CD25^＋Treg术后各时间点的表达分别为（3．36±0．29）％、（4．09±0．44）％、（5．44±0．35）％、（5．73±0．53）％，显著高于实验组的（2．50±0．39）％、（3．24±0．25）％、（4．20±0．45）％、（4．18±0．14）％，差异有统计学意义（t=3．828、2．898、3．780、4．892，均P〈0．05），两组脾脏中CIN^＋CD25^＋Treg的表达上调先于外周血；术后各时间点实验组外周血内CD8^＋CD103^＋Treg的表达分别为（2．20±0．33）％、（2．79±0．57）％、（4．55±1．03）％、（4．31±0．07）％，显著高于对照组的（0．73±0．12）％、（1．10±0．19）％、（1．43±0．14）％、（2．10±0．14）％，差�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, γ-interferon （IFN-γ） and IL-1β 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 （ χ^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

关 键 词：小鼠 角膜移植 移植物排斥 T淋巴细胞 调节性 细胞因子类 

分 类 号：R686[医药卫生—骨科学]