Exosome介导的T细胞信息传递在口腔扁平苔藓免疫炎性反应中的作用及机制研究

Oral lichen planus (OLP) is a chronic T cell-mediated immune and inflammatory disease. Exosome plays a key role in antigen presentation, T cell immune and inflammatory response, and keratinocyte apoptosis. Thus, it is speculated that exosome, via mediating signal transmission in T cells, might affect the function of T cell and biology of keratinocyte in OLP. (1) To detect the differential expression of immunity and inflammation associated gene and miRNA in T cell -derived exosome of OLP. (2) To analyze the variation of exosome and the clinical significance in OLP. (3) To build an OLP T cell-exosome model. (4) To examine the effect of the altered inflammation and immunity associated gene in exosome from T cell of OLP on T cell biology as well as T cell-mediated inflammation and immunity function. (5) To test the effect on T cell of immunity and inflammation associated miRNA in T-cell derived exosome on T cell of OLP. (6) To assay the role of exosome in the crosstalk between T cell and keratinocytes of OLP. (7) To investigate the related signalling pathways of exosome from T cell in OLP. (8) To explore the effect of exosome-based immune modulation on T cell in OLP. From the perspective of exosome-mediated signal transmission in T cells, this project potentially provides experimental basis for studying the immune inflammatory mechanism and exploring biological markers and targeted therapy for OLP, which has important scientific significance and promising application prospects.

口腔扁平苔藓（OLP）是T细胞介导的慢性免疫炎性疾病，外泌体可调控抗原提呈、T细胞的免疫炎性反应和角质形成细胞的凋亡。本项目提出OLP中的外泌体介导T细胞信息传递，导致OLP中T细胞的功能异常及角质形成细胞生物学特性的改变，从而影响OLP的发生发展。拟检测 OLP T细胞源性外泌体免疫炎症相关基因和miRNA的差异表达谱；分析OLP外泌体变化规律及临床意义；构建OLP T细胞-外泌体模型；观察免疫炎症相关基因沉默前后，OLP T细胞外泌体对T细胞生物学特性及免疫炎性功能的影响；研究OLP T细胞外泌体免疫炎症相关miRNA对T细胞的调控作用；探讨外泌体在OLP T细胞和角质形成细胞相互对话中的作用；探究OLP T细胞外泌体的相关信号通路；阐述基于外泌体的免疫调节在OLP T细胞中的作用。本项目为研究OLP的免疫炎性机制、探索其生物标记物和靶向治疗提供实验依据，具有重要科学意义和应用前景。

口腔扁平苔藓（OLP）是一种常见的由T细胞介导的免疫炎症性疾病。外泌体在多种炎症性疾病和自身免疫性疾病的发病机制中发挥重要作用。本项目旨在研究外泌体在OLP中的作用及调控机制。.主要研究内容:（1）探究OLP患者循环外泌体对T细胞生物学功能的影响。（2）应用miScript® miRNA PCR Array对OLP患者循环外泌体中的miRNA进行差异筛选，RT-PCR验证差异筛选结果，分析其临床意义。（3）分离OLP T细胞源性外泌体，检测OLP T细胞源性外泌体对T细胞增殖的影响，采用Luminex液相芯片筛选外泌体刺激后差异表达的细胞因子。（4）探讨差异表达的细胞因子MIP1-α/β对OLP T细胞的功能及作用机制。（5）构建T细胞源性外泌体-角质形成细胞共培养模型，探究OLP T细胞源性外泌体对角质形成细胞凋亡的影响。检测OLP T细胞源性外泌体中差异表达的细胞因子水平。.研究结果：（1）糜烂型OLP 循环外泌体促进T细胞的增殖、DNA的合成和细胞迁移，抑制T细胞凋亡。（2）OLP循环外泌体中miR-34a-5p的表达水平显著升高，与OLP病情正相关。miR-34a-5p的功能主要集中在生物学过程方面，可能通过作用于PI3K/Akt通路参与OLP进程。（3）OLP患者T细胞源性外泌体可刺激Jurkat细胞产生MIP-1α/β。MIP-1α/β在OLP病损组织和患者血浆中的含量明显升高，其受体CCR1和CCR5在OLP T细胞表面显著表达，且CCR1/5+ T细胞大多数为CD8+T细胞。（4）MIP-1α/β可促进PBMCs的迁移，抑制CCR1和CCR5可显著降低PBMCs迁移，尤其是减少CD8+ T细胞的迁移。（6）OLP和对照组T细胞源性外泌体均可造成角质形成细胞的形态改变和凋亡。OLP外周血T细胞源性外泌体中IL-1β、IL-5、IL-10、IL-12、IFN-γ、MIP-1β显著降低，IL-17A显著升高。两组外泌体均高表达TNF-α，这可能是角质形成细胞凋亡的原因。.本研究提示OLP循环外泌体影响T细胞的增殖、细胞周期、迁徙和凋亡。OLP循环外泌体中差异表达的miR-34a-5p与OLP病情相关，OLP T细胞源性外泌体可通过分泌MIP-1α/β诱导CD8+ T细胞迁移，还可诱导角质形成细胞凋亡。本研究揭示了外泌体在OLP免疫发病机制中的重要作用。