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。

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 the lentivirus. The titer of the recombinant lentivirus was 1.0×10⁶TU/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-γ 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 occurring and proceeding of TAO in mice.