长链非编码RNA NeST调控非节段型白癜风CD8+CTLs细胞IFN-γ通路活化的机制研究

中文摘要

英文摘要

Global activation of CD8+ cytotoxic T lymphocytes (CTLs), which release increased level of IFN-γ leading to apoptosis and destruction of melanocytes, plays a pivotal role in non-segmental vitiligo (NSV) patients. Yet, the mechanisms underlying the activation of CD8+ CTLs in NSV patients are unknown. Long noncoding RNA (lncRNA) plays a critical role in immune response. However, its role in the development of NSV is still undefined. Our research is based on the previous result from gene microarray expression chips and analysis of Gene Anotation Database, in which lncRNA NeST was validated to be involved in the activation of CD8+ CTLs. In this program, we will explore the differential expression of NeST in CD8+ T cells from peripheral blood or skin in a larger sample of NSV patients and normal controls, and delineate the correlation between the expression of NeST and the level of IFN-γ. Further, we will establish two overexpressing-NeST cell models, by which to explore the role of lncRNA NeST in regulating production of IFN-γ during the activation of CD8+ CTLs and discuss the underlying mechanism. This study will be expected to provide a novel targets for clinical prevention and treatment of NSV.

结题摘要

非节段型白癜风（NSV）患者存在系统性CD8+细胞毒性T淋巴细胞（CTLs）异常活化，产生大量IFN-γ引起皮肤黑素细胞凋亡和功能破坏，其分子机制尚不清楚。长链非编码RNA(lncRNA)参与调控机体的免疫反应，但其在NSV CD8+CTLs异常活化中的作用不明。本研究基于前期基因表达微阵列芯片技术和基因功能注解数据库，拟筛选出与NSV进展期CD8+CTLs活化相关的lncRNA NeST，扩大样本研究NeST在NSV患者与正常对照者外周血和皮疹来源CD8+T细胞中的表达差异，及其与IFN-γ的相关性。我们发现，lncRNA NeST在正常对照者、稳定期NSV和进展期NSV患者外周血CD8+T细胞中表达依次升高，在三组皮肤来源的CD8+T细胞中表达同样渐次上升，并且NeST的表达与IFN-γ+CD8+CTLs亚群比例呈正相关。体外实验中，我们使用原代人外周血 CD8+T 细胞构建过表达 lncRNA NeST细胞模型。经检测发现，过表达lncRNA NeST可刺激CD8+T细胞增殖，使得CD8+T淋巴细胞表面CD69和 CD44的阳性率增高，同时可导致活化的 CD8+T淋巴细胞分泌更多的 IFN-γ和IL-2，并具有一定抑制CD8+T细胞凋亡的作用。我们由此得出结论，lncRNA NeST 可刺激CD8+T细胞异常活化，生成更多IFN-γ，从而抑制黑素细胞功能，并可直接诱导黑素细胞凋亡，这可能是自身免疫因素参与白癜风发病的机制之一。本研究为自身免疫因素参与白癜风发病提供了新的证据，为白癜风的免疫学治疗提供了科学依据和潜在作用靶点。