国际眼科杂志
國際眼科雜誌
국제안과잡지
INTERNATIONAL JOURNAL OF OPHTHALMOLOGY
2013年
11期
2182-2184
,共3页
刘先宁%武鹏安%吴洁%程燕%潘士印%肖湘华%艾婷婷%安娜%王伟%朱秀萍
劉先寧%武鵬安%吳潔%程燕%潘士印%肖湘華%艾婷婷%安娜%王偉%硃秀萍
류선저%무붕안%오길%정연%반사인%초상화%애정정%안나%왕위%주수평
异种角膜移植%排斥反应%石蜡包埋组织%间接免疫荧光染色%CD4,CD8T淋巴细胞
異種角膜移植%排斥反應%石蠟包埋組織%間接免疫熒光染色%CD4,CD8T淋巴細胞
이충각막이식%배척반응%석사포매조직%간접면역형광염색%CD4,CD8T림파세포
heterologous cornea transplantation%rejection%paraffin embedding organizations%indirect immunofluorescence staining%CD4 and CD8 T lymphocytes
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， CD8 T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1，2 wk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1 wk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。
目的：採用單剋隆抗體間接免疫熒光法測定異種角膜移植術後不同時間點角膜組織中CD4， CD8 T淋巴細胞的變化。方法：取正常兔角膜、腋窩淋巴結組織；鴕鳥-兔異種角膜移植術後1，2 wk角膜組織，經甲醛固定，常規石蠟包埋切片脫蠟複水，胃酶消化法進行抗原脩複，用鼠抗兔CD4，CD8單剋隆抗體及熒光標記的羊抗鼠IgG進行免疫組化測定，熒光顯微鏡下觀察角膜組織中CD4＋， CD8＋T淋巴細胞的錶達。結果：鴕鳥-兔異種角膜移植後1 wk角膜組織中CD4＋， CD8＋T淋巴細胞的錶達為陰性，2wk時T淋巴細胞CD4呈彊暘性錶達，CD8呈陰性錶達。結論：採用間接免疫熒光法測定動物異種角膜移植術後不同時間點跼部角膜組織中CD4＋，CD8＋T淋巴細胞的錶達，為異種角膜移植排斥反應機製研究及術後藥物篩選奠定實驗基礎。
목적：채용단극륭항체간접면역형광법측정이충각막이식술후불동시간점각막조직중CD4， CD8 T림파세포적변화。방법：취정상토각막、액와림파결조직；타조-토이충각막이식술후1，2 wk각막조직，경갑철고정，상규석사포매절편탈사복수，위매소화법진행항원수복，용서항토CD4，CD8단극륭항체급형광표기적양항서IgG진행면역조화측정，형광현미경하관찰각막조직중CD4＋， CD8＋T림파세포적표체。결과：타조-토이충각막이식후1 wk각막조직중CD4＋， CD8＋T림파세포적표체위음성，2wk시T림파세포CD4정강양성표체，CD8정음성표체。결론：채용간접면역형광법측정동물이충각막이식술후불동시간점국부각막조직중CD4＋，CD8＋T림파세포적표체，위이충각막이식배척반응궤제연구급술후약물사선전정실험기출。
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.
