国际眼科杂志
國際眼科雜誌
국제안과잡지
INTERNATIONAL JOURNAL OF OPHTHALMOLOGY
2014年
2期
225-228
,共4页
陈华新%于少硕%周维明%张意
陳華新%于少碩%週維明%張意
진화신%우소석%주유명%장의
AHNAK1%T 细胞%RNA 干扰%甲状腺相关性眼病%免疫调控
AHNAK1%T 細胞%RNA 榦擾%甲狀腺相關性眼病%免疫調控
AHNAK1%T 세포%RNA 간우%갑상선상관성안병%면역조공
AHNAK1%T cells%RNA interference%thyroid-associated ophthalmopathy%immunoregulation
目的：构建表达小鼠CD4＋T细胞钙支架蛋白AHNAK1的短发夹RNA（short hairpin RNA ，shRNA）慢病毒载体，并研究其对小鼠甲状腺相关性眼病（ thyroid-associated ophthalmopathy ，TAO）的抑制效应。<br> 方法：设计并筛选对AHNAK1具有良好干扰效力的shRNA序列，慢病毒载体包装干扰序列，感染小鼠CD4＋T细胞，检测AHNAK1静默对T细胞功能的抑制作用，采用实验动物模型观察AHNAK1体内抑制甲状腺相关性眼病的效果。<br> 结果：成功筛选出具有良好干扰效力的shRNA ，并包装入慢病毒。病毒滴度为1．0×106 TU/mL，转染慢病毒的CD4＋T细胞展现出失能倾向，抑制炎症免疫反应；在动物模型中抑制T细胞中AHNAK1表达可以有效控制甲状腺眼病的发生发展，显著降低治疗组T细胞中IL-2、IL-1β和IFN-γ的表达。<br> 结论：成功构建了表达小鼠AHNAK1 shRNA的慢病毒，具有抑制T细胞分泌IL-2、IL-1β和IFN-γ的表达效应，能够有效抑制甲状腺眼病的发生发展。
目的：構建錶達小鼠CD4＋T細胞鈣支架蛋白AHNAK1的短髮夾RNA（short hairpin RNA ，shRNA）慢病毒載體，併研究其對小鼠甲狀腺相關性眼病（ thyroid-associated ophthalmopathy ，TAO）的抑製效應。<br> 方法：設計併篩選對AHNAK1具有良好榦擾效力的shRNA序列，慢病毒載體包裝榦擾序列，感染小鼠CD4＋T細胞，檢測AHNAK1靜默對T細胞功能的抑製作用，採用實驗動物模型觀察AHNAK1體內抑製甲狀腺相關性眼病的效果。<br> 結果：成功篩選齣具有良好榦擾效力的shRNA ，併包裝入慢病毒。病毒滴度為1．0×106 TU/mL，轉染慢病毒的CD4＋T細胞展現齣失能傾嚮，抑製炎癥免疫反應；在動物模型中抑製T細胞中AHNAK1錶達可以有效控製甲狀腺眼病的髮生髮展，顯著降低治療組T細胞中IL-2、IL-1β和IFN-γ的錶達。<br> 結論：成功構建瞭錶達小鼠AHNAK1 shRNA的慢病毒，具有抑製T細胞分泌IL-2、IL-1β和IFN-γ的錶達效應，能夠有效抑製甲狀腺眼病的髮生髮展。
목적：구건표체소서CD4＋T세포개지가단백AHNAK1적단발협RNA（short hairpin RNA ，shRNA）만병독재체，병연구기대소서갑상선상관성안병（ thyroid-associated ophthalmopathy ，TAO）적억제효응。<br> 방법：설계병사선대AHNAK1구유량호간우효력적shRNA서렬，만병독재체포장간우서렬，감염소서CD4＋T세포，검측AHNAK1정묵대T세포공능적억제작용，채용실험동물모형관찰AHNAK1체내억제갑상선상관성안병적효과。<br> 결과：성공사선출구유량호간우효력적shRNA ，병포장입만병독。병독적도위1．0×106 TU/mL，전염만병독적CD4＋T세포전현출실능경향，억제염증면역반응；재동물모형중억제T세포중AHNAK1표체가이유효공제갑상선안병적발생발전，현저강저치료조T세포중IL-2、IL-1β화IFN-γ적표체。<br> 결론：성공구건료표체소서AHNAK1 shRNA적만병독，구유억제T세포분비IL-2、IL-1β화IFN-γ적표체효응，능구유효억제갑상선안병적발생발전。
AIM: To construct the mice model with lentivirus expressing AHNAK1 shRNA in CD4+T cells, and to study its inhibitory effect on the thyroid - associated ophthalmopathy ( TAO) in mice. <br> METHODS:The shRNA sequence with good disturbing potency towards AHNAK1 was designed and selected;then the shRNA was packed into lentivirus;and the CD4+T cells were infected.The infected CD4+T cells of mice by the packed lentivirus were observed to detect the inhibition effect on T cells. And then the immunotherapeutic effects of AHNAK1-/-on TAO were observed by experimental animal model. <br> RESULTS: The shRNA with good disturbing potency was successfully screened and correctly inserted into the lentivirus.The titer of the recombinant lentivirus was 1.0 ×106 TU/mL.The CD4+T cells infected by lentivirus showed the anergy trend and restrained the immune response of inflammation.Suppressing the expression of AHNAK1 in T cells of animal model can effectively control the occurring and proceeding of TAO, which can significantly reduce the expression of IL-2 /IL-1β/IFN-γin the T cells of the control group. <br> CONCLUSION:This paper successfully constructs mice model with the recombinant lentivirus expressing AHNAK1 shRNA, which has a favorable inhibitory effect on secretion of IL-2, IL-1β, IFN-γby T cells.The recombinant lentivirus can effectively inhibit the occurring and proceeding of TAO in mice.
