Detecting Epstein-Barr virus DNA from peripheral blood mononuclear cells in adult patients with systemic lupus erythematosus in Taiwan

Epstein-Barr virus (EBV) has been found by many serology studies to be associated with systemic lupus erythematosus (SLE). However, the results of DNA studies have been conflicting. Therefore, instead of antibody to EBV, we studied the association between EBV DNA and SLE. In this case-control study in Taiwan, we enrolled 87 SLE patients and 174 age- and sex-matched controls. Peripheral blood mononuclear cells of SLE patients and matched controls were tested for EBV DNA by polymerase chain reaction (PCR) and Southern blot. Of the 87 SLE patients, 71 (81.6%) were found to be positive for EBV DNA, while 85 (48.9%) of the 174 controls (odds ratio 4.64, 95% confidence interval 2.50–8.62, P<0.0001) were positive. While the EBV DNA-positive rate did not decline with age in SLE patients (P>0.05), it did decline with age in controls (P<0.05). Furthermore, based on a real-time quantitative PCR study, we have found a significant difference between EBV viral load in SLE and controls (P=0.008). Therefore, in our molecular study of DNA level, we found evidence for the association of EBV infection and SLE, suggesting that EBV contributes, if not to the development of SLE, then to disease perpetuation.     

系统性红斑狼疮患者外周血白细胞介素15的研究

目的：检测系统性红斑狼疮（SLE）患者血清白细胞介素15（IL-15）水平及外周血单个核细胞（PBMC）IL-15mRNA表达，并进一步分析其临床意义。方法：IL-15检测采用ELISA方法；PBMCIL-15mRNA表达采用原位杂交法检测。结果：①SLE组患者血清IL-15水平显著高于正常对照组（P〈0．01），活动期SLE患者血清IL-15水平显著高于缓解期患者（P〈0．05）。②发生临床肾损害者IL-15水平明显高于无肾损害者（P〈0．05），出现血清抗dsDNA抗体阳性、低补体C3血症、高IgG血症者血清IL-15水平均分别显著高于无上述表现者。③SLE患者PBMCIL-15mRNA表达量明显高于正常对照组（P〈0．05），活动期SLE显著高于缓解期（P〈0．05）。④SLE患者PBMC培养上清IgG、IgM和抗dsDNA抗体浓度均显著高于正常对照组；SLE患者PBMCIL-15mRNA表达量与细胞培养上清的IgG及抗dsDNA抗体滴度均呈正相关关系（分别为r=0．645和r=0．715，P〈0．05），而与IgM．无相关关系（r=0．451，P〉0．05）。⑤SEE患者PBMCIL-15mRNA表达量与血清IL-15水平呈正相关关系（r=0．726，P〈0．05）。结论：SLE患者存在外周血IL-15蛋白和基因表达异常，且与其分泌免疫球蛋白和自身抗体有关，提示IL-15可能参与SLE的病理生理过程。     

外周血CD19+B细胞PD-L1的表达与系统性红斑狼疮发病的相关性分析

目的:探讨程序性死亡配体1(Programmed death ligand-1,PD-L1)在系统性红斑狼疮(Systemic lupus erythema-tosus,SLE)患者外周血B细胞上的表达及临床意义。方法:应用流式细胞仪检测51例SLE患者和38例健康对照者外周血CD19+B细胞表面PD-L1的表达水平,比较SLE稳定组、活动组和健康对照组以及狼疮肾炎组和无狼疮肾炎组之间CD19+B细胞表面PD-L1表达阳性细胞的百分比,并分析其与临床表现及实验室检查数据的相关性。结果:SLE活动组和稳定组CD19+PD-L1+B细胞百分率均低于健康对照组,活动组又低于稳定组,差异均有统计学意义(均P<0.05)。狼疮肾炎患者CD19+PD-L1+B细胞百分率低于无狼疮肾炎患者(P<0.05)。SLE患者CD19+PD-L1+B细胞百分率与SLEDAI评分、尿蛋白定量、呈负相关,与C3呈正相关。SLE患者中抗dsDNA抗体、抗Sm抗体、抗U1snRNP抗体、抗核小体抗体阳性组外周血B细胞PD-L1表达水平均低于对应阴性组,且均有统计学意义(均P<0.05)。结论:SLE患者外周血CD19+B细胞表达PD-L1下降,与病情活动性和抗体产生有很好的相关性。     

Expression of lncRNA NEAT1 in peripheral blood mononuclear cells of patients with systemic lupus erythematosus and its correlation with Th1/Th2 balance

Objective: This study explored and analyzed the expression of LncRNA NEAT1 in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE) and its correlation with Th1/Th2 balance. Methods: We chose 97 SLE patients admitted in our hospital from Jun. 2016 to Feb. 2019 as SLE group, and randomly selected 50 healthy volunteers that underwent physical examination in our hospital during the same period as control group. We detected the expression of LncRNA NEAT1 in PBMCs of the two groups of subjects by qRT-PCR, the degree of Th1 and Th2 cells in both groups by flow cytometry, and the expression of TFN-γ and IL-4 in both groups by ELISA. Results: The relative expression of LncRNA NEAT1 in PBMCs of SLE group was higher than that of control group (P<0.05). The proportion of Th1 and the ratio of Th1/Th2 cells in PBMCs were markedly lower in the SLE group than the control group (P<0.05), while the proportion of Th2 was higher in the SLE group than the control group (P<0.05). IFN-γ level in SLE group was much lower than the control group (P<0.05), while IL-4 level was evidently higher in the SLE group than in controls (P<0.05). The expression of LncRNA NEAT1 in PBMCs of SLE group was notably negatively correlated with Th1 proportion and Th1/Th2 ratio (P<0.05), while positively correlated with Th2 proportion (P<0.05). Conclusion: LncRNA NEAT1 in PBMCs of SLE patients is abnormally highly expressed, and this expression is negatively correlated with Th1/Th2 balance. These two factors may interact and jointly affect the occurrence and progression of SLE.     

系统性红斑狼疮患者外周血单个核细胞miR-125b及miR-145的表达及临床意义

目的探讨miR-125b及miR-145在系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中的表达水平及其临床意义。方法选取海南省人民医院收治的136例SLE患者,采用SLE病情活动指数(SLEDAI)评分将其分为低活动组(n=75)和高活动组(n=61),另选取60例健康体检者作为对照组。采用实时定量PCR(RT-PCR)法检测各组PBMC中miR-125b及miR-145水平,分析其对SLE患者的诊断价值。Pearson相关分析SLE患者miR-125b及miR-145水平与抗双链DNA(dsDNA)抗体、C3、C4、IgG、血沉(ESR)、C反应蛋白(CRP)及SLEDAI评分的相关性。结果 SLE组、高活动组和低活动组抗dsDNA抗体、ESR、IgG及CRP水平均明显高于对照组(P<0.05),而C3及C4水平均明显低于对照组(P<0.05)。SLE组、高活动组和低活动组PBMC中miR-125b(0.84±0.23、0.42±0.09和1.53±0.28 vs 6.12±1.83)及miR-145(0.68±0.17、0.35±0.06和1.12±0.20 vs 2.15±0.64)表达水平均明显低于对照组(P<0.01),且高活动组PBMC中miR-125b(0.42±0.09 vs 1.53±0.28)及miR-145(0.35±0.06 vs1.12±0.20)表达水平明显低于低活动组(P<0.01)。SLE患者治疗后miR-125b(2.60±0.71 vs 0.84±0.23)及miR-145(1.96±0.58 vs 0.68±0.17)表达水平明显高于治疗前(P<0.01),SLE患者治疗后抗dsDNA抗体(96.24±17.20 vs 272.36±64.35)水平明显低于治疗前(P<0.01)。ROC曲线分析显示,miR-125b、miR-145及抗dsDNA抗体三项联合诊断SLE的AUC(95%CI)为0.928(0.871～0.986),其敏感度(90.4%)和特异度(85.0%)较好。相关分析显示,SLE患者miR-125b、miR-145与抗dsDNA抗体、IgG、SLEDAI评分均呈负相关(P<0.05),miR-125b与miR-145呈明显正相关(r=0.805,P<0.05)。结论 miR-125b及miR-145水平在SLE患者PBMC中明显降低,且与SLE病情活动相关,联合抗dsDNA抗体检测对SLE诊断具有一定价值。     

SLE患者外周血单个核细胞分泌白细胞介素6和IgG的研究

我们研究了21例SLE患者和正常人外周血单个核细胞(PBMC)体外培养自发和丝裂原刺激后细胞增殖、IL—6产生、IgG分泌的情况。发现SLE 患者PBMC经PHA和PWM刺激后,细胞增殖的指数降低;用IL—6细胞依赖株(MH_(60)·BSF_2)检测培养上清液中的IL—6,发现活动期SLE患者PBMC自发和PHA刺激后产生的IL—6明显高于缓解期SLE患者(P<0.001)和正常人(P<0.001);活动期和缓解期SLE患者PBMC体外自发分泌IgG量增高,分别为正常人自发分泌量的5.68倍和4.48倍;我们首次用统计学方法分析了SLE患者细胞培养上清液中的IL—6水平与分泌的IgG量的关系,发现两者有显著的相关性(r=0.7046,P<0.001)。     

系统性红斑狼疮患者外周血单个核细胞转录因子NF—AT和AP—1活性的初步观察

目的观察系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)NF-AT和AP-1活性,并进一步探讨其临床意义.方法NF-AT和AP-1活性检测采用电泳迁移率改变法(EMSA).结果①活动期(19例)和缓解期(13例)SLE患者PBMC NF-AT活性均显著高于正常对照组(P＜0.05);活动期SLE显著高于缓解期(P＜0.05).②活动期SLE组PBMC AP-1活性均显著低于正常对照组(P＜0.01);缓解期SLE与正常对照组无明显差别(P＞0.05).③血清抗dsDNA抗体阳性SLE患者(21例)NF-AT明显高于抗dsDNA抗体阴性组(11例)(P＜0.05),而两组患者AP-1活性无显著差异(P＞0.05).④SLE患者PBMC NF-AT活性与CRP和SLEDAI呈正相关关系,与ESR、ANA、C3无相关关系;AP-1与上述各临床指标无相关关系.结论SLE外周血单个细胞中存在介导细胞内信号转导的转录因子NF-AT和AP-1表达异常,这可能与SLE的发病机制有关;NF-AT可能与SLE的疾病活性有关.     

Ets-1在系统性红斑狼疮中的研究进展

系统性红斑狼疮(SCE)是一种典型的系统性自身免疫性疾病,其发病机制尚未明确,其中T、B细胞功能异常起着重要作用.转录因子Ets-1作为SLE的易感基因之一在淋巴细胞分化与细胞因子调节上起重要作用.Ets-1除影响B细胞分化和功能外,对T细胞的生存、增殖、发育和功能起重要作用.虽然Ets-1在SLE发病中的确切机制仍尚未明确,但越来越多的研究表明Ets-1在SLE的发生发展中起着重要作用.     

系统性红斑狼疮患者外周血Th1细胞自噬相关标记物的表达

目的:探讨系统性红斑狼疮(Systemic lupus erythematosus,SLE)患者外周血Th1细胞自噬相关标记物的表达情况及意义.方法:应用流式细胞术检测68例SLE患者及23例正常人外周血Th1细胞自噬相关标记物的表达水平,并分析其与补体C3 、SLEDAI评分和血清Anti-dsDNA水平的相关性.结果:SLE患者Th1细胞自噬标记物Beclin-1和Phospho-mTOR的水平均低于正常人,而LC3的水平高于正常人(均P＜0.05);活动期SLE患者Phospho-mTOR水平低于非活动期患者(P＜0.05).SLE患者Th1细胞Phospho-mTOR水平与SLEDAI评分呈负相关,与血清C3水平呈正相关,与血清Anti-dsDNA水平呈负相关(均P＜0.05).外周血Th1细胞Phospho-mTOR和补体C3水平变化在判断SLE患者疾病活动情况时敏感性高于血清Anti-dsDNA,但特异性不如血清Anti-dsDNA.结论:SLE患者外周血Th1细胞存在自噬异常现象,且自噬相关标记物Phos-pho-mTOR的表达与病情活动度相关,有望成为评价SLE活动度的指标之一.     

系统性红斑狼疮病人血T,B淋巴细胞Bcl—2的表达

目的：探讨Ｂｃｌ－２在系统性红斑狼疮（ＳＬＥ）发病机制中作用。方法：采用流式细胞仪双标记法检测３１例ＳＬＥ病人外周血Ｔ、Ｂ细胞Ｂｃｌ－２蛋白表达。结果：发现活动期ＳＬＥ病人活动期ＳＬＤ病人ＣＤ３＾＋、ＣＤ４＾＋和ＣＤ８＾＋Ｔ细胞Ｂｃｌ－２蛋白表达明显高于非活动期ＳＬＥ病人、其他疾病组和正常对照组。ＣＤ１９＾＋Ｂ细胞Ｂｃｌ－２蛋白表达在各组之间并无统计学差异。ＣＤ３＾＋Ｔ细胞Ｂｃｌ－２蛋白表达的平均     

Activation of IL-6 production by UV irradiation of blood mononuclear cells from patients with systemic lupus erythematosus.

Cultured mononuclear cells from patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and normal donors were assayed for their ability to secrete IL-6 both spontaneously and after exposure to UV light. Mononuclear cells from SLE, RA and atopic control patients produced IL-6 spontaneously, while those from normal donors did not. Spontaneous production of IL-6 occurred in the non-adherent cell population. UV light-induced IL-6 production was confined exclusively to the SLE patients and was present only in the macrophage/monocyte fraction. This stimulation was induced by wavelengths in the UVA, UVB but not the UVC portion of the spectrum. These results suggest that cytokine release may be involved in the exacerbations of SLE provoked by photosensitivity.     

钙调神经磷酸酶-活化T细胞核因子与系统性红斑狼疮

系统性红斑狼疮是一种以多种自身抗体产生为特点的侵犯多系统多脏器的自身免疫性疾病,其发病机制尚未明确,目前认为T淋巴细胞异常是SLE的发病的关键.钙调神经磷酸酶-活化T细胞核因子信号通路作为T细胞内重要的生物信号转导通路,在T细胞活化中起到调节枢纽的作用,了解CaN/NFAT信号途径在SLE中的作用及目前相关主要治疗药物的研究现状可为SLE的基础研究及临床治疗提供依据.     

系统性红斑狼疮患者外周血单个核细胞中急性期蛋白反应因子的表达

目的　探讨系统性红斑狼疮 (SLE)患者外周血单个核细胞 (PBMC)中急性期蛋白反应因子 (APRF)的活性水平 ,以及IL 6和IL 10对APRF表达的影响。方法　采用凝胶阻滞电泳 (EMSA)的方法检测 4 0例SLE患者及 2 0例正常对照组PBMC中DNA结合蛋白APRF的表达水平。结果　所有活动期SLE患者均出现APRF电泳条带 ,17例非活动期SLE患者中有 10例出现APRF条带 ,而正常人对照组无 1例出现。 7例未出现APRF电泳条带的非活动期SLE患者PBMC加IL 10处理后均出现不同程度的APRF表达 ,而加IL 6处理时仍未出现APRF电泳条带。结论　SLE患者存在APRF的异常表达。在SLE中 ,IL 10信号转导途径中的某些调控机制 (如蛋白激酶 )可能发生改变 ,从而使得核内的APRF激活转录 ,提示IL 10很可能是通过APRF在SLE的发病机制中起作用 ,相反IL 6在SLE发病的作用机制很可能与APRF无关。     

雷公藤内酯醇对系统性红斑狼疮患者DC细胞功能及成熟的影响

目的 通过体外实验研究雷公藤内酯醇(triptolide)对系统性红斑狼疮(systemic lupus erythematosus,SLE)患者树突状细胞(dendritic cell,DC)功能及成熟的影响,为进一步阐明雷公藤内酯醇的免疫学活性提供依据.方法 从SLE患者外周血分离单个核细胞,流式细胞仪分选DC,加入0、5、10、30μg/L的雷公藤内酯醇共孵育,24h后收集上清液,ELISA检测IFN-α、IL-6、TNF-α量,5d后收集细胞,流式细胞仪检测DC表型CD11c、CD80、CD86阳性率,光镜观察DC的形态,扫描电镜观察DC的超微结构.结果 雷公藤内酯醇显著减低活动期与非活动期SLE患者IFN-α、IL-6、TNF-α量,并呈雷公藤内酯醇浓度依赖性(P＜0.05);雷公藤内酯醇可抑制SLE患者DC的分化和成熟,并呈雷公藤内酯醇浓度依赖性(P＜0.05).结论 雷公藤内酯醇能够减弱SLE患者DC的功能,并抑制其分化和成熟.     

Phenotype and function of dendritic cells of patients with systemic lupus erythematosus

Dendritic cells (DC) regulate the activation and differentiation of T cells. They are activated by signals of inflammation and tissue damage, and thus could play a role in the amplification and perpetuation of autoimmune diseases such as systemic lupus erythematosus (SLE). Here we analyzed the phenotype of circulating DC from patients with SLE and studied their differentiation from monocytes. Peripheral blood DC exhibited increased levels of activation in patients with SLE. Although their in vitro differentiation process occurred normally, their responses to activation stimuli (LPS, TNF-α plus PGE(2), anti-CD40) were abnormal when compared to DC differentiated from healthy monocytes. When incubated in the presence of IL-10, DC from patients with SLE were able to induce tolerance to allogeneic antigens in a normal manner. Our results suggest that DC from patients with SLE are abnormal, in part due to the effect of abundant pro-inflammatory signals, but also because of intrinsic cellular defects that alter their responses to activation stimuli.     

系统性红斑狼疮患者外周血CD4+CD25high Tr和 CD4+CD25low T细胞PD-1表达的分析和意义

目的 探讨PD-1(programmed death-1)在系统性红斑狼疮(systemic lupus erythematosus,SLE)患者外周血CD4+ CD25high调节性T细胞(regulatory T cell,Tr)和CD4+ CD25lowT细胞上的表达及临床意义.方法 应用流式细胞仪检测51例SLE患者和38例健康对照者外周血CD4+ CD25high Tr和CD4+ CD25lowT细胞表面PD-1表达水平,同时收集SLE患者临床表现、实验室检查数据,比较SLE稳定组、活动组和健康对照组以及狼疮肾炎组和无狼疮肾炎组之间CD4+ CD25 high Tr和CD4+CD25lowT细胞表PD-1表达的百分比,并分析其与临床表现及实验室检查数据的相关性.结果 (1)SLE活动组和稳定组外周血CD4+ CD25 high Tr和CD4+ CD25lowT细胞的百分率均低于对照组.(2)SLE活动组和稳定组外周血CD4+ CD25 highTr表达PD-1的百分率均高于对照组.SLE稳定组和对照组外周血CD4+ CD25lowT细胞表达PD-1的百分率均高于活动组.(3)狼疮肾炎患者外周血CD4+CD25highTr表达PD-1的百分率高于无狼疮肾炎患者.而狼疮肾炎患者CD4+ CD25lowT细胞表达PD-1的百分率低于无狼疮肾炎患者.(4) CD4+CD25high PD-1+T细胞百分率与SLEDAI评分呈正相关;CD4+ CD25high Tr、CD4+CD25lowPD-1+T细胞百分率与SLEDAI评分呈负相关.结论 SLE患者外周血中CD4+ CD25high Tr、CD4+ CD25lowT细胞以及它们表面PD-1表达异常,并且与疾病活动性相关,PD-1/PD-L1信号通路可能对CD4+CD25highTr、CD4+CD25lowT细胞的数量和功能有影响.