IL-10基因修饰脂肪来源间充质干细胞治疗自身免疫性干眼机制研究

Autoimmune related dry eye is one of most common eye diseases, which can significantly affect vision acuity and the quanlity of life. Our preliminery data showed that adipose-derived mesynmensal stem cells (ADSCs) could effectively inhibit the development of autoimmune dry eye in rabbit. Based on our preliminery data and the latest progression in this field, this project will explore the therapeutic effects of IL-10-ADSCs in autoimmune dry eye in rabbit induced by transfer of antigen-specific T cells. We will analyze the effcts of IL-10-ADSCs injected at different time points on the autoimmune dry eyes. The immunomodulatory mechanisms of the ADSCs will be elucidated by analyzing the changes of the function and frequency of the three T cell subsets:Th1 Th17 and Tregs, and by exploring the effects of ADSCs-secreted cytokines on them, especially on the balance beteween the Th17 and Treg cells.The migratory capacity of IL-10-ADSCs will also be studied. This project will futher optimize the therapeutic strategy of ADSCs by investigating the possible mechanism of treating autoimmune dry eye with IL-10-ADSCs, and will provide strong experimental basis for the future clinical application in ophthalmology.

自身免疫相关性干眼严重影响人们的生活质量，寻找安全有效的治疗方法是临床亟待解决的问题。我们的前期研究结果表明脂肪间充质干细胞（ADSCs）可以有效抑制自身免疫性干眼的发生和发展，治疗可能与上调调节性T细胞相关细胞因子IL-10的表达有关。结合前期的研究结果和最新的进展，本课题将通过兔特异性T细胞诱导的兔自身免疫性干眼模型，研究IL-10基因修饰的脂肪间充质干细胞（IL-10-ADSCs）在不同的给药模式和时机下，对干眼的治疗及对致病性T细胞致病能力的影响；并通过检测淋巴器官和泪腺Th1、Th17和Treg细胞数目和功能变化，以及IL-10-ADSCs对T细胞亚群的影响，尤其是对Th17和Treg平衡的影响，阐明IL-10-ADSCs在自身免疫性干眼中的免疫调节机理；初步探索IL-10-ADSCs治疗干眼中的定向迁移能力及机制。从中找到新靶点，为自身免疫性干眼的预防和新药开发提供依据。

间充质干细胞(Mesenchymal stem cells, MSCs)的免疫调节特性使其在自身免疫性干眼的治疗中具有较大的应用前景，近年来研究表明，用特定的免疫调节因子对MSCs进行基因修饰，联合基因治疗和细胞治疗的优势，可显著提高其治疗作用。本研究成功构建了IL-10 (Interleukin-10, IL-10) 基因修饰的脂肪来源间充质干细胞 (IL-10-ADSCs)，利用兔自身免疫性干眼模型证实了其对自身免疫性干眼的保护作用显著优于ADSCs，机制研究表明，IL-10-ADSCs在体内体外均可抑制滤泡辅助T细胞(Tfh)的分化与功能，其对Tfh细胞的调控作用部分归因于抑制miR-142-5p的表达。本研究为进一步优化MSCs治疗自身免疫性干眼的临床应用提供了新的思路和实验基础。