系统性红斑狼疮患者外周血单个核细胞中白细胞介素-10mRNA表达的研究

目的 探讨白细胞介素(IL-10)在系统性红斑狼疮(SLE)中的作用。方法 采用逆转录多聚酶链反应(RT-PCR)及酶联免疫吸附法(ELISA)测定40例SLE患者和20例正常对照组外周血单核细胞(PBMC)IL-10mRNA表达及IL-10自发分泌水平。结果 SLE患者PBMC自发分泌IL-10水平及其IL-10mRNA表达水平均显著高于正常对照组(P<0.01),其中SLE活动期明显高于非活动期(P<0.01),而非活动期又明显高于正常对照组(P<0.01)。结论 IL-10在SLE发病中起重要作用,PBMC分泌IL-10水平对SLE诊断和病情活动性监测有重要临床意义,拮抗SLE患者体内IL-10水平,将为SLE治疗开辟一条新途径。     

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.     

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。     

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。     

Abnormal CTLA‐4 function in T cells from patients with systemic lupus erythematosus

CTLA‐4 is a critical gatekeeper of T‐cell activation and immunological tolerance and has been implicated in patients with a variety of autoimmune diseases through genetic association. Since T cells from patients with the autoimmune disease systemic lupus erythematosus (SLE) display a characteristic hyperactive phenotype, we investigated the function of CTLA‐4 in SLE. Our results reveal increased CTLA‐4 expression in FOXP3^? responder T cells from patients with SLE compared with other autoimmune rheumatic diseases and healthy controls. However, CTLA‐4 was unable to regulate T‐cell proliferation, lipid microdomain formation and phosphorylation of TCR‐ζ following CD3/CD28 co‐stimulation, in contrast to healthy T cells. Although lupus T cells responded in vitro to CD3/CD28 co‐stimulation, there was no parallel increase in CTLA‐4 expression, which would normally provide a break on T‐cell proliferation. These defects were associated with exclusion of CTLA‐4 from lipid microdomains providing an anatomical basis for its loss of function. Collectively our data identify CTLA‐4 dysfunction as a potential cause for abnormal T‐cell activation in patients with SLE, which could be targeted for therapy.     

Regulatory T cells in systemic lupus erythematosus

Systemic lupus erythematosus (SLE), an autoimmune disease, develops when immunologic self‐tolerance fails. Treg cells are a subset of CD4⁺ T cells that maintain self‐tolerance by suppressing autoreactive lymphocytes. Defects in Treg cells are therefore considered to be an aspect of SLE pathogenesis. Nevertheless, reports on the numbers and function of Treg cells in SLE are contradictory and the definitive role of Treg cells in SLE remains unclear. In this review, we summarize findings from murine models and ex vivo experiments, which provide insights into the mechanisms that result in the breakdown of tolerance. We also include recent findings about Treg‐cell subsets and their markers in human SLE. The identification of unique markers to identify bona fide Treg cells, as well as therapies to reconstitute the balance between Treg cells and autoreactive T cells in SLE, are the future challenges for SLE research.     

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。     

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。     

A comprehensive review of the clinical approach to pregnancy and systemic lupus erythematosus

Nowadays, most of the young women affected by Systemic Lupus Erythematosus (SLE) can carry out one or more pregnancies thanks to the improvement in treatment and the consequent reduction in morbidity and mortality. Pregnancy outcome in these women has also greatly improved in the last decades. A correct timing for pregnancy (tailored on disease activity and established during a preconception counselling), together with a tight monitoring during the three trimesters and the post-partum period (to timely identify and treat possible obstetric complications or maternal disease flares), as well as the concept of multidisciplinary management, are currently milestones of the management of pregnancy in SLE patients. Nevertheless, the increasing knowledge on the compatibility of drugs with pregnancy has allowed a better treatment of these patients, by choosing medications that control maternal disease activity without harming the foetus. However, particular attention and strict monitoring should be dedicated to SLE pregnant women in particular clinical settings: patients with lupus nephritis and patients with aPL positivity or Antiphospholipid syndrome, who are at higher risk for maternal and foetal complications, but also patients with anti-Ro/SSA and/or anti-La/SSB antibodies, because of the risk of neonatal lupus. A discussion on family planning, as well as counselling on contraception, should be part of the everyday-practice for physicians caring for SLE women during their reproductive age. Another issue is the possible reduction of fertility in these women, that can be due to different reasons. Consequently, the request for assisted reproduction techniques has been increasing in the last years, so that rheumatologists and gynaecologists should be prepared to counsel SLE patients also in this particular setting.     

Increased frequency of activated regulatory T cells in patients with lupus nephritis

Background and ObjectiveLupus nephritis (LN) is one of the common manifestations of systemic lupus erythematosus (SLE), affecting the quality of life of patients. Abnormality in the adaptive immune response, such as T cell response, plays the main role in the pathogenesis of SLE and LN. In this study, we aimed to evaluate the role of different subpopulations of regulatory T cells (Tregs) and effector T cells (Teff) in LN patients and compare them with SLE patients.Materials and MethodsA total of 48 participants were enrolled in this study and divided into 3 groups: (i) patients with SLE; (ii) patients with LN; and (iii) healthy controls (HCs). The frequencies of CD4⁺ CD25⁺⁺ CD45RA^? Foxp3^hi activated Tregs (aTregs), CD4⁺ CD25⁺ CD45RA⁺ Foxp3^lo resting Tregs (rTregs), CD4⁺ CD25⁺ CD45RA^? Foxp3^lo non-Tregs, CD4⁺ CD25⁺ Foxp3^? Teff, and Tregs were analyzed in all subjects using a flow cytometer.ResultsLN patients had a significantly increased frequency of aTregs and Tregs compared with SLE patients (standardized mean difference [SMD] = 0.50; 95% CI [-0.26, 1.25]; p > 0.05 and SMD = 0.60; 95% CI [-0.16, 1.36]; p > 0.05, respectively). Patients with LN had a significantly increased frequency of Teff compared with SLE patients (SMD = 0.49; 95% CI [-0.26, 1.24]; p > 0.05). However, an increased ratio of Tregs/Teff was observed in LN patients compared with SLE patients (SMD = -0.25; 95% CI [-0.97, 0.48]; p > 0.05).ConclusionPatients with LN showed immunoregulatory properties, in which both aTregs and Tregs had increased frequencies.     

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     

T cells in the pathogenesis of systemic lupus erythematosus

Recent studies in patients with systemic lupus erythematosus (SLE) have demonstrated that autoantigen-reactive T cells can be isolated from peripheral blood and that such cells can support autoantibody production ex vivo, suggesting that they may have a central role in the pathogenesis of disease. In addition, recent work has identified and characterized signaling abnormalities in T cells from SLE that may be fundamental to the disease. This review will examine the role of T cells in the pathogenesis of SLE and it will consider pathogenic mechanisms by which T cells escape normal of immunological tolerance. The focus will be on recent studies characterizing autoantigen-reactive human T cells and signaling abnormalities identified in T cells from patients with SLE.     

系统性红斑狼疮外周血单个核细胞CD40L的表达增高

目的:了解系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMCs)的白细胞分化抗原40配体(CD40L)表达,探讨其在发病中的作用。方法:分离SLE患者和正常人PBMCs,采用流式细胞术,检测其在正常状况和应用植物凝集素(PHA)及地塞米松(Dex)后,CD40L的表达水平,并进行比较;分析SLE患者CD40L的表达水平和狼疮活动指数(SLEDAI)的相关性。结果:活动期SLE患者PBMCs的CD40L阳性细胞百分率(%)明显高于对照组,且高于静止期SLE患者;应用PHA处理24h后,3组PBMC表达CD40L均明显增加,但活动期SLE患者增加更明显;应用地塞米松后,SLE患者(活动期和静止期)PBMCs的CD40L表达明显减少,对照组无明显改变;SLE患者(活动期和静止期)CD40L的表达水平和SLEDAI均呈明显正相关。结论:CD40L在SLE患者PBMCs的表达增加,和疾病活动度有关;其受PHA和Dex调控,在SLE发病和病程中起重要作用。     

血清抗C1q抗体在系统性红斑狼疮疾病活动性判断及狼疮肾炎诊断中的价值

目的探讨抗C1q抗体(C1qAb)在系统性红斑狼疮(SLE)活动性及狼疮肾炎(LN)诊断和疾病活动性判断中的价值。方法采用酶联免疫吸附法检测SLE患者(n=89)、疾病对照组(n=56)和正常对照组(n=42)血清中的抗C1q抗体阳性率,并与SLE患者临床实验室指标﹑活动性评分进行分析。结果 C1qAb的阳性率在SLE患者中显著高于疾病对照组和正常对照组患者(P<0.05);C1qAb阳性的SLE患者肾损发生率、活动性狼疮发生率及抗dsDNA抗体的阳性率均高于C1qAb阴性患者(P<0.05);C1qAb与SLEDAI活动性评分、抗核小体抗体(anti-nucleosome antibody,AnuA)及抗dsDNA抗体呈正相关(P<0.05)。结论抗C1q抗体对SLE的诊断和疾病活动性判断有重要价值;抗C1q抗体参与了SLE肾脏损害的发病机制。     

Matrix metalloproteinase-9 and its natural inhibitor TIMP-1 expressed or secreted by peripheral blood mononuclear cells from patients with systemic lupus erythematosus

Matrix metalloproteinase-9 (MMP-9) was involved in inflammation and immune system dysfunctions. Besides immunologic abnormalities, systemic lupus erythematosus (SLE) also presents chronic inflammatory components. Therefore, a role of MMP-9 in SLE pathology might be supposed. To verify this hypothesis, SLE patients and healthy donors were compared for the MMP-9 and MMP-9 mRNA levels in peripheral blood mononuclear cells (PBMCs), the spontaneous secretion of MMP-9 and TIMP-1 and the MMP-9 activity. Thus, we found that fresh PBMCs from SLE patients expressed a significantly higher activity of MMP-9 and spontaneously released higher levels of MMP-9, as compared to healthy donors, while the secreted TIMP-1 level was the same for both groups. When the patients were sub-grouped based on disease status, the most increased pro-MMP-9 activity inside the PBMCs was identified for relapse SLE sub-group. A similar observation for SLE patients with positive serum fibrinogen was found. Following culture, the PBMCs from remission SLE patients secreted significantly higher MMP-9 level, than the PBMCs from relapse SLE patients. PBMCs from relapse SLE patients secreted the highest levels of TIMP-1, although this difference was not statistically significant. Taken together, these observations suggested the multiple roles of MMP-9 and TIMP-1 in progress of inflammation and tissue damage and/or in repair, depending on clinical stages of SLE.     

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中IL—4和IL—13 mRNA的表达

目的探讨Th2型细胞因子在系统性红斑狼疮(SLE)发病机制中的作用及其意义.方法应用逆转录-聚合酶链反应(RT-PCR)检测了34例活动期SLE患者和30例正常人外周血单个核细胞(PBMC)中IL-4和IL-13mRNA的表达水平.结果活动期SLE患者IL-4和IL-13的阳性表达率与正常人对照组相比均无明显差异(P>0.05);活动期SLE患者PBMC中IL-4和IL-13mRNA的平均表达水平(0.938 6±0.168 9,0.898 3±0.115 3)均明显高于正常人对照组(0.5494±0.151 0,0.608 5±0.090 3),差异非常显著(P<0.001).结论Th2型细胞因子IL-4和IL-13在SLE患者中呈高水平表达,这可能与SLE患者外周血T细胞高度活化、功能异常有关.