shRNA静默CD4~＋ T细胞AHNAK1基因对小鼠甲状腺相关性眼病进程的抑制作用  

Inhibition effect of the thyroid-associated ophthalmopathy in mice through the lentivirus expressing AHNAK1 shRNA on T cells

作　　者：陈华新[1] 于少硕[2] 周维明[3] 张意[3] 

机构地区：[1]广州军区武汉总医院眼科,中国湖北省武汉市430070 [2]第二军医大学研究生管理大队临床九队,中国上海市200433 [3]第二军医大学免疫学教研室,中国上海市200433

出　　处：《国际眼科杂志》2014年第2期225-228,共4页International Eye Science

基　　金：国家自然科学基金青年项目(No.30801000)~~

摘　　要：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。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。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1.0×106TU/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。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。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, 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. RESULTS： The shRNA with good disturbing potency was successfully screened and correctly inserted into thelentivirus. The titer of the recombinant lentivirus was 1.0xl08TU/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- II3/IFN-（ in the T cells of the control group. 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-γ by T cells. The recombinant lentivirus can effectively inhibit the occurrinq and oroceedina of TAO in mice.

关 键 词：AHNAK1 T细胞 RNA干扰 甲状腺相关性眼病 免疫调控 

分 类 号：R771.3[医药卫生—眼科]