ALA/SFC诱导的DCreg对移植免疫耐受的诱导效果及其作用机制研究

Dendritic cells (DCs) play a key role in cellular immune response. In recent years, some research had found that DC was a group of heterogeneous cells. Some subclasses of DCs have a negative regulatory function, which were referred to as regulatory DC. And the regulatory DC plays an important role in peripheral regulatory T-cell induction and maintenance. 5-aminolevulinic acid (ALA) is widely used as an amino acid photodynamic cancer diagnosis and treatment. Our previous study has firstly proved that ALA/SFC induce MHC mismatched donor mice cardiac transplantation produce specific transplantation tolerance effect. And we also found a special phenomenon in the receptor of cardiac transplant models with a large amount of the regulatory DC. Therefore, in this study we will focus on the methods and properties of regulatory DC induced by ALA/SFC and the mechanism of in vivo induction of allograft tolerance effect using in vitro culture, flow cytometry, mixed lymphocyte experiment, mice heart transplantation, adoptive cell experiments, RT-PCR, cell antibody screening, protein staining and so on. From three dimensions- cells, molecules and, RNA to clarify ALA/SFC induce allograft tolerance mechanism. This study for the first time will provide a scientific explanation of allograft tolerance induced by ALA/SFC via elevated regulatory DC.

调节性DC在外周T细胞耐受诱导和维持中发挥重要作用。5-氨基酮戊酸(ALA)作为一种光动力药物,其产物CO及胆红素/胆绿素被证实具有抑制免疫排斥反应作用。我们先期研究发现ALA具有抑制淋巴细胞增殖作用,并首次发现ALA联合二价铁枸橼酸亚铁钠（SFC）具有诱导MHC错配的小鼠心移植产生供体特异性的移植耐受,并诱发受体产生大量调节性DC的特有现象。本申请课题中，我们(1)将利用ALA/SFC体外培养获得调节性DC。通过培养供体和受体的DCreg和传统培养方法比较其生物学活性。(2)通过心移植模型输注供体或者受体的DCreg验证其诱导免疫耐受效果。(3)通过抗体阻断,细胞移除，细胞过继实验探讨DCreg诱导免疫耐受机制,绘制其作用机制路线,从细胞、分子和RNA三个层面来阐明ALA/SFC诱导同种移植耐受的作用机制,亦为今后继续研发这种安全有效与传统免疫抑制剂作用机制不同的新型免疫调节药物奠定基础

调节性DC在外周T细胞耐受诱导和维持中发挥重要作用。5-氨基酮戊酸(ALA)作为一种光动力药物,其产物CO及胆红素/胆绿素被证实具有抑制免疫排斥反应作用。我们(1)将利用ALA/SFC体外培养获得调节性DC。通过培养供体和受体的DCreg和传统培养方法比较其生物学活性。(2)通过心移植模型输注供体或者受体的DCreg验证其诱导免疫耐受效果。(3)通过抗体阻断,细胞移除，细胞过继实验探讨DCreg诱导免疫耐受机制,绘制其作用机制路线,从细胞、分子和RNA三个层面来阐明ALA/SFC诱导同种移植耐受的作用机制,亦为今后继续研发这种安全有效与传统免疫抑制剂作用机制不同的新型免疫调节药物奠定基础。体外研究部分我们顺利完成，获得了相应的结果并且写成相关文章《5-ALA联合SFC体外诱导小鼠骨髓来源调节性树突状细胞的培养与鉴定》发表在《中华器官移植杂志》上。心移植模型上的研究，数据收集整理后我们准备参与Frontiers in Immunology 杂志2022的专题Novel Therapies for Tolerance Induction in Solid Organ and Bone Marrow Transplantation: Volume II组稿文章。同时我们也进行了相关的研究如睾丸热损伤模型，小鼠抑郁模型及急性腹膜炎模型上利用ALA+SPF进行治疗，取得了良好的结果，相关实验数据我们整理后将进行发表。同时，我们进行了糖尿病相关勃起功能障碍的大鼠模型上进行了尿源性干细胞分泌的外泌体相关治疗研究，取得了非常好的结果，相关结果已经投稿于stem cell research and therapy。同时，相关外泌体在心移植方面的治疗也在进行中。