负载供肾抗原的受者大鼠树突状细胞诱导肾移植免疫耐受性研究

It became a prospective therapeutics that Dendritic cells (DCs) can induce transplantation immune tolerance for its bi-directional immune regulatory function. In our study we have successfully obtained recipient rats DC2 in vitro, and then confirmed that it was able to regulate Th1/Th2 immune response and induce the specific recipient-to-donor immune tolerance in the kidney transplantation model. However, its detailed mechanism remains unclear. And during the process, we further found the upregulation of phosphorylated STAT6 levels and the downregulation of phosphorylated STAT4 levels. Therefore, we speculated that DC2 induced immune tolerance through STAT6/CCL17 and STAT4/ NF-κB/IL-12 pathways. Based on these results, this project will intend to focus on studying STAT-related signaling pathways. So, at the molecular level, we will fully screen these signaling molecules and regulatory pathways which may be involved by the use of liquid-chip technology; and then in the kidney transplantation model, we will perform siRNA or overexpression experiments to verify the role of these molecular pathways in immune tolerance induced by DC2, and establish evaluation indexes to characterize the effect, then further clarify the signaling pathway and influence in the induction of donor-specific immune tolerance by recipient DC2 loaded with donor kidney antigens. In all, this project will not only provide a new basis for the theory of induced specific immune tolerance in kidney transplantation, and also, lay a new foundation for the clinical application of DC2 induced adoptive immunotherapy in transplantation.

树突状细胞（DC）因其双向免疫调节功能而成为诱导移植免疫耐受的有效手段。我们在体外成功制备了大鼠DC2，并在肾移植模型中证实它能调控Th1/Th2型免疫应答，诱导受者产生对供体的特异性免疫耐受，但其机制尚不清楚。我们又发现，此过程中DC2的STAT6和STAT4的磷酸化水平分别有升高和下降，说明其诱导免疫耐受可能与STAT6/CCL17和STAT4/NF-κB/IL-12等调控通路相关。据此，本项目拟围绕STAT相关调控通路，在分子层面用液态芯片技术全面筛查其中可能涉及的信号分子和调控通路；在肾移植模型层面，用siRNA或过表达技术验证这些分子通路在DC2诱导免疫耐受过程中的作用，并建立考察指标表征其效果，从而阐明负载供肾抗原的受者DC2诱导肾移植免疫耐受信号通路及其对耐受性影响。本项目研究将为肾移植中诱导特异性免疫耐受的理论提供新依据，为DC2诱导免疫过继疗法在移植中的应用打下新基础。

树突状细胞（DC）因其双向免疫调节功能而成为诱导移植免疫耐受的有效手段。我们在体外成功制备了大鼠DC2，并在肾移植模型中证实它能调控Th1/Th2 型免疫应答，诱导受者产生对供体的特异性免疫耐受，但其机制尚不清楚。据此，本项目围绕STAT 相关调控通路，在分子层面用液态芯片技术全面筛查其中可能涉及的信号分子和调控通路；在肾移植模型层面，用siRNA 或过表达技术验证这些分子通路在DC2 诱导免疫耐受过程中的作用，并建立考察指标表征其效果，从而阐明负载供肾抗原的受者DC2 诱导肾移植免疫耐受信号通路及其对耐受性影响。项目研究发现，负载供肾抗原的未成熟树突状细胞能有效抑制受体T细胞增生，吲哚胺2，3双加氧酶（IDO)在负载供肾抗原的受体表达明显提高，以及负载供体抗原未成熟树突状细胞(iDCs)和IDO信号通路有助于移植肾免疫耐受和延长其存活时间。本项目研究为肾移植中诱导特异性免疫耐受的理论提供了新依据，为DC2 诱导免疫过继疗法在移植中的应用打下新基础。