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

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-associatedophthalmopathy,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伊106TU/mL,转染慢病毒的CD4＋T细胞展现出失...

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Bibliographic Details
Published in国际眼科杂志 Vol. 14; no. 2; pp. 225 - 228
Main Author 陈华新 于少硕 周维明 张意
Format Journal Article
LanguageChinese
Published 广州军区武汉总医院眼科, 中国湖北省武汉市,430070%第二军医大学研究生管理大队临床九队, 中国上海市,200433%第二军医大学免疫学教研室, 中国上海市,200433 2014
Subjects
AHNAK1
RNA干扰
T细胞
免疫调控
甲状腺相关性眼病
甲状腺相关性眼病
T 细胞
免疫调控
RNA interference
AHNAK1
immunoregulation
thyroid-associated ophthalmopathy
RNA 干扰
T cells
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Summary: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-associatedophthalmopathy,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伊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-1茁和IFN-酌的表达。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-1茁和IFN-酌的表达效应,能够有效抑制甲状腺眼病的发生发展。
Bibliography:61-1419/R
Hua-Xin Chen, Shao-Shuo YU, Wei-Ming Zhou, Yi Zhang 1Department of Ophthalmology, Wuhan General Hospital of Guangzhou Military Command of Chinese PLA, Wuhan 430070, Hubei Province, China; 2 Post-graduate Administration Company, the Second Military Medical University, Shanghai 200433, China; 3 Department of Immunology, the Second Military Medical University, Shanghai 200433, China
AHNAK1; T cells; RNA interference;thyroid-associated ophthalmopathy; immunoregulation
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 observ
ISSN:1672-5123
DOI:10.3980/j.issn.1672-5123.2014.02.07