IL-10基因单核苷酸多态性与SLE患者免疫异常的相关性研究

High serum level of the cytokines interleukin-10 (IL-10) is an essential factor of immune injury and dysfunction in systemic lupus erythematosus (SLE) patients. mRNA and protein expression may be affected by genetic polymorphisms. Three common single nucleotide polymorphisms (SNPs): a G to A substitution at position −1082 (rs1800896), a C to T at −819 (rs1800871) and a C to A at −592 (rs1800872), are from the transcriptional start site in the 5′ flanking region of the IL-10 gene which is a strong determinant for IL-10 production in preliminary studies. Our previous study suggested that IL-10−1082G/A polymorphism was significantly associated with SLE susceptibility. However no correlation of −819C/T and −592C/A polymorphisms with SLE risk was found. hIL-10 gene has 184 SNPs. Some of these SNPs may upregulate the expression of IL-10 in SLE patient by gene transcription and translation process, resulting in the patient's immune abnormality. . In the present study, genomic DNA was extracted from peripheral blood samples of SLE patients and controls stored in our laboratory. Genotyping for these 184 SNPs are to be performed by Fluidigm SNPtype assays. Single factor logistic regression will be used to confirm that which SNPs are associated with SLE susceptibility. Furthermore,the SNP-SNP interactions in these SLE susceptibility loci are to be tested with multifactor dimensionality reduction (MDR) statistical method.. From January 2016 to December 2018, peripheral venous blood will be collected from new-onset SLE patients and controls. To detect the lever of IL-10, the separated serum for enzyme-linked immunosorbent assay is to be performed. Total RNA is to be extracted from PBMC using TRIzol Reagent. Quantitative real-time PCR is to be performed to detect IL-10 mRNA using a 7300 Real-Time PCR System. Genotyping for SNPs vulnerable to SLE are to be performed by Fluidigm SNPtype assays. Then, the appropriate statistical analysis methods will be conducted. Based on the above research, this study will illustrate IL-10 SNPs for the mechanism of action of SLE immune abnormality.

SLE免疫异常与IL-10升高密切相关。IL-10基因某些单核苷酸变异(SNPs)，如1082G>A可改变相关转录因子致IL-10水平升高；课题组前期研究发现IL-10-1082G/A与SLE发病相关。hIL-10基因184个SNPs，哪些影响IL-10表达，与患者免疫异常相关，未见系统性报道。课题拟用Fluidigm基因分型技术检测实验室备存病例和对照标本184个SNPs基因型，进行单因素Logistic回归及多因子降维分析，明确与SLE相关IL-10 SNPs及交互作用；采用Fluidigm基因分型、RT-PCR、ELISA和流式细胞术分别检测2016.1-2018.12采集标本的SNPs基因型、IL-10 mRNA、血清IL-10和免疫细胞生物标志分子，探讨IL-10 SNPs与SLE免疫异常相关性；建立IL-10 SNPs与患者临床表型数据库，为SLE风险预测和临床诊疗提供指导。

系统性红斑狼疮患者中 IL-10水平升高 ，可能是患者免疫损害及功能失调发生的重要原因，在疾病发生与发展中发挥重要作用，但SLE 发病相关的 IL-10 SNPs 及其交互作用，以及它们与 SLE 患者免疫异常的相关性未见深入研究。本研究采用病例-对照研究方法，筛选与 SLE 易感相关的 IL-10 SNPs，检测SLE患者和健康对照血浆IL-10水平和IL-10 mRNA表达情况，并探讨IL-10 mRNA和血浆 IL-10 表达水平与SLE 患者免疫异常的关系。研究发现SLE两个新待确认的易感位点，IL-10基因rs3024498多态性与SLE易感性相关，部分SNPs基因分型或等位基因与临床表现相关；SLE患者IL-10水平和IL-10 mRNA表达水平高于对照，SLE 患者IL-10 mRNA 表达水平与血浆IL-10 水平无相关性。研究为相关动物实验研究奠定了理论与实验基础，为临床 SLE 风险预测和个体化治疗提供了理论依据。