IL-35在结节病发病机制中的研究

发布时间：2018-04-27 14:18

本文选题：结节病 + 白介素35　；参考：《北京协和医学院》2017年博士论文

【摘要】：背景和目的:我们先前研究已经发现,Treg/Th17失衡及Treg细胞免疫抑制功能缺陷是结节病发病的重要原因之一。IL-35作为Treg细胞免疫抑制功能的执行者,通过促进Treg细胞增殖,抑制Th17分化并诱导iTr35细胞(分泌IL-35的Treg细胞亚群)生成发挥免疫抑制作用。我们推测结节病中Treg细胞的抑制功能缺陷和Treg/Th17失衡可能与IL-35分泌减少或其在细胞内信号传导途径异常有关。因此本研究将在先前研究基础上明确结节病中合成和释放IL-35的细胞来源及其表达情况;探讨IL-35在结节病的发病机制中可能发挥的生物学效应,此研究旨在探寻结节病的发病机制和寻找潜在的治疗靶点。方法:ELISA检测结节病组(n=98)和正常对照组(n=98)血浆中IL-35的水平,Real-time PCR检测结节病组和正常对照组IL-35两个亚基EBI3和p35的表达水平,以及Treg细胞转录因子Foxp3的表达水平。Pearson相关分析结节病患者血浆中IL-35的水平与临床参数的相关性。选取初治结节病患者10例,健康对照者10例,采用流式细胞术检测外周血中Treg细胞占CD4+T细胞的比例,IL-35在CD4+T细胞、CD8+T细胞、Treg细胞上的表达情况。结果:结节病组血浆中IL-35的水平(43.56±11.77pg/mL)明显低于正常对照组(55.18± 11.53 pg/mL),差异具有统计学意义(P0.001)。结节病治疗组(n=57)较未治疗组(n=41)血浆中IL-35的水平明显升高(47.38±12.02pg/mL vs 40.82± 10.89 pg/mL;P=0.0059),结节病患者血浆中IL-35水平与DLCO占预测值%成正相关(r=0.76,P0.001),而与其他临床参数未发现明显相关性。结节病组EBI3和Foxp3基因mRNA的表达水平均明显低于正常对照组(分别为 P=0.0016,P=0.0391),而p35基因mRNA的表达水平在两组间无明显差别(P =0.5816),结节病组和正常对照组EBI3基因相对表达量与Foxp3基因相对表达量均成明显正相关(分别为 r =0.786,P0.001;r =0.730,P0.001),p35 与 Foxp3 基因相对表达量在结节病组成低度正相关(r =0.383,P =0.037),而在正常对照组两基因表达无明显相关性(r=0.208,P =0.271)。流式细胞术检测发现,10例结节病患者外周血中Treg/CD4+T细胞的平均比例(5.82±0.51%)与健康对照者(5.71 ±0.90%)相比无显著差异(P=0.7413),在CD4+T细胞、CD8+T细胞和Treg细胞中均未发现表达IL-35的细胞。结论:结节病患者血浆中IL-35水平较正常健康者明显减低,且经激素和或免疫抑制剂治疗后水平升高,提示IL-35可能参与了结节病的炎症过程,在其发病过程中具有重要作用。结节病患者血浆中IL-35水平与DLCO占预测值%成正相关,提示其有可能作为预测结节病肺部病变进展的生物学指标。IL-35的一个亚基EBI3和Treg细胞转录因子Foxp3的mRNA表达水平均明显低于正常对照组,且EBI3、p35基因与Foxp3基因相对表达量成正相关,提示结节病患者外周血中Treg细胞数量的减少可能导致了 EBI3、p35基因生成减少,以至于EBI3与p35形成的异源二聚体减少,最终合成IL-35减少。而本研究采用流式细胞术检测人类T淋巴细胞亚群并未发现IL-35表达阳性的细胞,可能与人T淋巴细胞亚群中分泌IL-35的细胞数量太少有关,抑或IL-35可能主要由其它细胞亚群所分泌。
[Abstract]:Background and purpose: our previous studies have found that the Treg/Th17 imbalance and Treg cell immunosuppressive function defects are one of the important causes of sarcoidosis,.IL-35 as the executor of the immunosuppressive function of Treg cells, by promoting the proliferation of Treg cells, inhibiting the differentiation of Th17 and inducing the generation of iTr35 cells (the Treg cell subgroup of IL-35) to produce hair. We speculate that the inhibitory functional defects of Treg cells and Treg/Th17 imbalance in sarcoidosis may be related to the decrease of IL-35 secretion or the abnormal signal transduction pathway in the cells. Therefore, this study will make clear the synthesis and release of IL-35 cells and their expression in sarcoidosis based on previous studies, and discuss IL-35 in the study. The biological effects may be played in the pathogenesis of sarcoidosis. This study aims to explore the pathogenesis of sarcoidosis and to find potential therapeutic targets. Methods: ELISA detection of IL-35 levels in plasma of sarcoidosis group (n=98) and normal control group (n=98), Real-time PCR to detect EBI3 and p35 of IL-35 two subunits in nodal disease group and normal control group. The level of expression and the expression level of Treg cell transcription factor Foxp3 were correlated with the correlation between the level of IL-35 in the plasma of sarcoidosis patients and the correlation of clinical parameters. 10 patients with primary sarcoidosis and 10 healthy controls were selected, and the proportion of Treg cells to CD4+T cells in peripheral blood was detected by flow cytometry, and IL-35 in CD4+T cells, CD8+T. Result: the level of IL-35 in the plasma of the sarcoidosis group (43.56 + 11.77pg/mL) was significantly lower than that of the normal control group (55.18 + 11.53 pg/mL), and the difference was statistically significant (P0.001). The level of IL-35 in the plasma of sarcoidosis group (n=57) was significantly higher than that in the untreated group (n=41) (47.38 + 12.02pg/mL vs 40.82 + 10.89 P. G/mL; P=0.0059), the level of IL-35 in the plasma of sarcoidosis patients was positively correlated with the predicted value of DLCO (r=0.76, P0.001), but not significantly correlated with other clinical parameters. The expression level of EBI3 and Foxp3 gene mRNA in sarcoidosis group was significantly lower than that of normal control group (P= 0.0016, P=0.0391), while the expression level of p35 gene was between the two groups. There was no significant difference (P =0.5816). The relative expression of EBI3 gene in the sarcoidosis group and the normal control group was positively correlated with the relative expression of the Foxp3 gene (R =0.786, P0.001; R =0.730, P0.001), and the relative expression of p35 and Foxp3 genes in the low degree of sarcoidosis, but in the normal control group two gene table. There was no significant correlation (r=0.208, P =0.271). Flow cytometry showed that the average proportion of Treg/CD4+T cells in peripheral blood of 10 sarcoidosis patients (5.82 + 0.51%) was not significantly different from that of healthy controls (5.71 + 0.90%) (P=0.7413). No IL-35 cells were found in CD4+T cells, CD8+T fine cells and Treg cells. Conclusion: Sarcoidosis The level of IL-35 in the patient's plasma is significantly lower than that of the normal health, and the level of IL-35 may be involved in the inflammatory process of sarcoidosis, which may play an important role in the pathogenesis of sarcoidosis. The level of IL-35 in the plasma of sarcoidosis patients is positively correlated with the predictive value of DLCO, suggesting that it may be a precondition. The biological index of the progression of sarcoidosis pulmonary disease.IL-35, the mRNA expression level of a subunit EBI3 and Treg cell transcription factor Foxp3 was significantly lower than that of the normal control group, and the EBI3, p35 gene was positively correlated with the relative expression of Foxp3 gene, suggesting that the decrease of Treg cell in the peripheral blood of sarcoidosis patients may lead to EBI3, p35 basis. As a result of the reduction of the formation of the heterogenous two polymer formed by EBI3 and p35, the final synthesis of IL-35 decreased. In this study, the use of flow cytometry to detect human T lymphocyte subsets did not find IL-35 positive cells. It may be related to the small number of cells secreting IL-35 in human T lymphocyte subsets, or IL-35 may be mainly caused by others. Secreted by cell subgroups.

【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R593.9

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