BTLA联合CCR7基因共转染未成熟树突状细胞诱导皮肤移植免疫耐受的研究

How to prolong the mean survival time of alloskin transplantation is one of the main problems of large area burn wound repair. Previous studies have shown that CCR7 gene transfected into immature dendritic cells (imDCs) can enhance the migration ability of imDCs, but can promote imDCs differentiate towards maturation and influence the effect of immunologic tolerance induced by imDCs. How to make imDCs after transfected by CCR7 gene remains higher immune tolerance function is an urgent problem to be solved. BTLA is a negative inhibitory receptor, BTLA-HVEM combine with each other and plays negative feedback regulation role. The project intends to apply gene cotransfection technique, transfect CCR7 and BTLA genes into the donor imDCs, make the imDCs after transfection has automatic homing recipient lymph node and keep strong immune tolerance. Observation the morphology、phenotype、function and gene expression of imDCs before and after transfection in vitro, and observation the content of imDCs and the expression of signaling molecule in lymph node and the effect of skin allograft rejection in the mouse allogeneic skin transplantation model in vivo. The purpose of this project is to induce the imDCs developed more immune tolerance function, provide a new method for prolong the mean survival time of allogeneic skin transplantation after large area burn and how to induce immune tolerance.

如何延长同种异体皮肤存活时间是大面积烧伤创面修复领域的主要问题之一。前期研究证实CCR7基因转染未成熟树突状细胞(imDCs)能增强imDCs迁移能力,但可促使imDCs部分向成熟分化,影响诱导免疫耐受的效果。如何使CCR7基因转染后imDCs仍保持较强的免疫耐受功能是亟待解决的问题。BTLA为负性抑制性受体，BTLA-HVEM通过相互结合起着负反馈调节作用。本项目拟采用基因共转染技术,将CCR7和BTLA基因共转入供者源imDCs，使其具有自动归巢受者淋巴结又保持较强免疫耐受的功能。体外观察转染前后imDCs细胞形态、表型、功能以及相关基因的表达情况，体内观察小鼠异基因皮肤移植模型中imDCs在各淋巴结的含量、各信号通路分子表达情况以及对皮肤移植排斥反应的影响。本项目旨在诱导imDCs发挥更强的免疫耐受作用,为大面积烧伤治疗延长异基因皮肤移植存活时间及如何诱导免疫耐受提供一个新方法。