IL-36R及其通路对肠道GVHD免疫损伤修复作用的机制研究

Current studies found that the new proinflammatory factor of IL-36 has played an important role in the immune related diseases. The receptor of IL-36 (IL-36R) can can promote intestinal mucosa damage repair in inflammatory bowel disease. However, as the disease related with immune inflammatory, whether intestinal GVHD has the effect of regulating immune damage repair is not reported. Our previous work found that the expression of IL-36 in peripheral blood was higher in patients with intestinal GVHD, and the more seriously, the higher expression. Meanwhile, the expression of IL-36R after treatment of intestinal GVHD showed the down trends. Therefore, the development of intestinal GVHD may have the correlation with the IL-36R signalling. This study will use the animal model to validate the correlation between the IL-36R signalling and the intestinal GVHD from the level of mRNA, protein and cell. According to the IL-36R or MyD88 gene knockout mice transplant models, clearing that IL-36R intervened intestinal GVHD by MyD88 and its downstream regulation factors. We got the cells which are closely related to the intestinal mucosa damage, and observed the expression of IL-36R under the stimulus of IL-36 (the environment of GVHD) in vitro, monitored the cell proliferation activity, analysed the factors which were associated with the cells by next generation sequencing. Combining with clinical specimens to confirm that the function and mechanism of IL-36R signalling in mucosa damage repair in intestinal GVHD, and to provide the theory basis of discovering the potential therapeutic targets of intestinal GVHD.

近期研究发现促炎因子IL-36在免疫相关疾病中有重要作用，其受体IL-36R可促进炎性肠病粘膜修复，但对免疫炎性相关肠道GVHD是否也有免疫损伤修复作用目前未见报道。我们前期工作发现肠道GVHD患者IL-36表达升高，且严重度较高者更明显，治疗后出现下调；因此肠道GVHD可能与IL-36/IL-36R通路相关。本研究将利用动物移植模型，从mRNA、蛋白质及细胞水平验证IL-36R及通路与肠道GVHD相关性；通过IL-36R及MyD88基因敲除小鼠模型，明确IL-36R通过MyD88及下游调控因子干预肠道GVHD；获取与肠粘膜损伤修复相关细胞，体外实验观察其在IL-36刺激下（GVHD环境）IL-36R表达及细胞增殖活性，基因测序寻找相关因子，明确肠道GVHD损伤修复细胞及靶点；结合临床样本验证IL-36R及通路对肠道GVHD粘膜损伤修复免疫调控作用，为发现潜在肠道GVHD治疗靶点提供依据。

促炎因子IL-36在免疫相关疾病中有重要作用,其受体IL-36R可促进炎性肠病粘膜修复,但对免疫炎性相关肠道GVHD是否也有免疫损伤修复作用目前未见报道。前期工作发现肠道GVHD患者IL-36表达升高,考虑因此肠道GVHD可能与IL-36/IL-36R通路相关。本研究成功构建小鼠异基因造血干细胞移植后肠道GVHD模型,发现移植后小鼠肠道GVHD过程中相关炎性细胞因子特异表达，包括IL-36表达亦有特异改变。体外实验发现在活化T细胞刺激下(GVHD环境)IL-36表达有相似的特异性改变。收集临床异基因移植后肠道GVHD患者的外周血样本，发现在GVHD诊断及治疗后相关炎性因子，包括IL-36有均有变化，考虑IL-36在肠道GVHD的发生发展过程中有着重要作用。