荧光染色法在实验性异种角膜移植术后免疫诊断中的应用  

Application of paraffin embedding slice fluorescent staining method in the experimental xenogenic corneal transplantation immune diagnosis

作　　者：刘先宁 武鹏安 吴洁[1,4] 程燕[1,4] 潘士印[1,2] 肖湘华[1,2] 艾婷婷[1,2] 安娜[1,2] 王伟[3] 朱秀萍[1,2] 

机构地区：[1]中国陕西省西安市第一医院 [2]陕西省眼科研究所,710002 [3]中国陕西省西安市第一医院病理科,710002 [4]西安市眼科医院,710002

出　　处：《国际眼科杂志》2013年第11期2182-2184,共3页International Eye Science

基　　金：陕西省科技统筹创新工程计划项目(No.2012KTCQ03-11)~~

摘　　要：目的:采用单克隆抗体间接免疫荧光法测定异种角膜移植术后不同时间点角膜组织中CD4,CD8T淋巴细胞的变化。方法:取正常兔角膜、腋窝淋巴结组织;鸵鸟-兔异种角膜移植术后1,2wk角膜组织,经甲醛固定,常规石蜡包埋切片脱蜡复水,胃酶消化法进行抗原修复,用鼠抗兔CD4,CD8单克隆抗体及荧光标记的羊抗鼠IgG进行免疫组化测定,荧光显微镜下观察角膜组织中CD4+,CD8+T淋巴细胞的表达。结果:鸵鸟-兔异种角膜移植后1wk角膜组织中CD4+,CD8+T淋巴细胞的表达为阴性,2wk时T淋巴细胞CD4呈强阳性表达,CD8呈阴性表达。结论:采用间接免疫荧光法测定动物异种角膜移植术后不同时间点局部角膜组织中CD4+,CD8+T淋巴细胞的表达,为异种角膜移植排斥反应机制研究及术后药物筛选奠定实验基础。AIM ： Using monoclonal antibodies indirectly immunofluorescence to detect the changes of CD4 and CD8 T lymphocytes in the different time points in corneal tissue after corneal transplantation. METHODS： Normal rabbit cornea, axillary lymph node tissue were taken. Get corneal tissue at one week, two weeks after ostrich- rabbit heterogeneous corneal transplantation, fixed by formaldehyde, conventional paraffin embedding sectioning dewaxing after water, gastric enzyme digestion method for antigen repair, use mouse anti rabbit CD4, CD8 monoclonal antibody and fluorescent tagging sheep anti mouse IgG for immunohistochemical assay, and observe the expression of CD4~, CD8＋T lymphocytes in the cornea tissue in fluorescence microscope. ~ RESULTS： The expression of CD4~, CD8＊T cells in the cornea tissue was negative one week after the ostrich- rabbit heterologous cornea transplantation, CD4 T lymphocytes showed strong positive expression after the second week , CD8 was negative. = CONCLUSION： Indirect immunofluorescence method can be used for determination of related antigen expression in local corneal tissue after heterologous corneal transplantation, and it can lay the foundation for the studies of heterogeneous cornea transplantation rejection mechanism and postoperative drug screening.

关 键 词：异种角膜移植 排斥反应 石蜡包埋组织 间接免 疫荧光染色 CIM CD8T淋巴细胞 

分 类 号：R779.65[医药卫生—眼科]