系统性红斑狼疮患者干扰素诱导基因的表达及其与疾病活动性的相关性研究

目的 探讨系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMCs)中5个干扰素(IFN)诱导基因(IFIT1、IFIT4、OAS1、OASL、ISG15)的表达及其与SLE疾病活动度的相关性.方法 运用SYBRgreen dye I实时定量聚合酶链反应(RT-PCR)方法检测76例SLE患者与54名健康对照人的IFIT1、IFIT4、OASL、OASL及ISG15 mRNA表达水平(以-AACt值表示),并且与红细胞沉降率(ESR)、C反应蛋白(CRP)、补体C3、C4、抗核抗体(ANA)及抗dsDNA抗体等指标比较,分析IFIT1、IFIT4、OAS1、OASL、ISG15 mRNA表达水平以及上述常规检测指标与SLE疾病活动指数(SLEDAI)积分之间的相关性.结果 ①SLE组与正常对照组相比,IFIT1、IFIT4、OAS1、OASL及ISG15 mRNA表达显著增高(P＜0.01);SLE活动组与SLE缓解组比较,IFIT1、IFIT4、OAS1、OASL及ISG15 mRNA表达增高(P＜0.05);SLE组IFIT1、IFIT4、OAS1、OASL及ISG15 mRNA表达水平之间呈正相关(r＞0.5,P＜0.05).②SLE组IFIT1、IFIT4、OAS1、OASL及ISG15 mRNA表达水平均与SLEDAI积分呈显著正相关(r＞0.5,P＜0.05).③ESR、CRP、补体C3、C4、ANA与IFIT1、IFIT4、OAS1、OASL、ISG15 mRNA表达水平及SLEDAI积分均无相关性,抗dsDNA抗体与IFIT1、IFIT4、OAS1、OASL、ISG15 mRNA表达水平及SLEDAI积分呈正相关(r＞0.5,P＜0.05).结论 SLE患者IFIT1、IFIT4、OAS1、OASL及ISG15的表达水平在SLE患者中显著增高,并且与SLEDAI积分呈显著正相关,对SLE患者病情活动度和病情严重度的判断均有较大价值,抑制这5个基因的表达可能为SLE的治疗提供新的靶点.     

系统性红斑狼疮患者Ⅰ型干扰素诱导基因的表达

目的 通过观察系统性红斑狼疮(SLE)患者Ⅰ型IFN诱导基因[黏液病毒抗性蛋白1(MXI)、2',5'-寡腺苷酸样合成酶(OASL)、2',5'-寡腺苷酸合成酶1(OASI)、α-干扰素诱导蛋白15(ISGl5)、淋巴细胞抗原6复合体E(LY6E)]mRNA表达,探讨其对SLE活动或感染鉴别诊断的作用.方法 收集48例SLE患者、16例类风湿关节炎(RA)患者、26例健康人外周血细胞,提取RNA,用实时荧光定量PCR检测所有受试者MXI、OASL、OASI、ISG15、LY6E mRNA表达.单因素和多因素logistic回归分析上述5个基因表达水平与狼疮感染或活动的关联性.结果 (1)与健康人相比,SLE患者MXI、OASL、OASI、ISGl5、LY6E mRNA表达水平显著增高,差异有统计学意义(P值均小于0.01);SLE患者OASL、OAS1、ISG15、LY6E mRNA表达水平较RA者显著增高,差异有统计学意义(P值均小于0.01).(2)男性SLE患者MXI、OASL、OASI、ISGl5、LY6E mRNA表达水平与女性患者相比,差异均无统计学意义.(3)logistic回归分析显示,ISG15、LY6E是SLE活动的独立危险因素(P相似文献   

干扰素诱导基因与系统性红斑狼疮的相关性研究

目的 探讨实时定量测定干扰素诱导基因[抗黏病毒1(MX1)基因,2′5′-寡腺苷酸合成酶1(OAS1),干扰素诱导蛋白44(IFI44)基因]表达水平与系统性红斑狼疮(SLE)临床表现及病情活动度的相关性.方法 收集100例SLE患者,40例非SLE其他自身免疫性疾病患者,40名健康对照的临床资料,取外周血抽提总RNA并反转录成cDNA,运用SYBR green dye Ⅰ实时定量聚合酶链反应(QT-PCR)在ABI PRISM 7000基因测序仪上检测患者和对照组的MXI、OAS1和IFI44定量表达水平(以△CT值表示)的差异,并与各临床指标及病情活动度进行相关性分析.采用方差分析和Spearman相关分析进行统计学处理.结果 ①SLE患者总体的MX1、OAS1、IFI44 mRNA △CT值(3.4±1.8,4.2±1.5,8.8±2.2)明显高于非SLE患者组(2.4±0.4,3.4±0.7,5.4±2.1)和健康对照组(2.3±1.2,2.6±0.7,5.2±2.0);②SLE患者组的OAS1、IFI44 mRNA △CT值在不同活动度的SLE组内[非活动组(3.8±1.4,3.2±1.8);轻度活动组(4.8±1.5,8.0±2.2);重度活动组(6.0±1.4,12.1±2.4)]差异具有统计学意义;③SLE患者组的OAS1、IFI44 mRNA△CT值与SLE疾病活动指数(SLEDAI)积分之间有相关性(r=0.338,0.380);④在有关节炎的SLE患者中,其MX1、OAS1、IFI44表达水平升高,与无关节炎患者比较差异具有统计学意义;⑤在狼疮肾炎组内,IFI44mRNA △CT值表达(2.2±1.1)与非狼疮肾炎组(3.2±2.1)比较差异有统计学意义;⑥SLE组内3个基因表达存在高相关性且具有统计学意义(P均＜0.05).结论 干扰素诱导基因MX1、OAS1、IFI44在SLE患者中均有表达上调现象,OAS1、IFI44 mRNA实时定量表达水平对SLE患者的病情活动度判断有意义,且三者在SLE的发生发展中亦存在密切联系.     

OAS异构体对系统性红斑狼疮合并感染或疾病活动鉴别诊断的初步研究

目的探讨2′,5′-寡腺苷酸合成酶(OAS)异构体对系统性红斑狼疮(SLE)活动或感染的鉴别诊断作用。方法用荧光定量聚合酶链反应(PCR)检测SLE活动组(34例),SLE合并感染组(28例),正常对照组(29名)的OAS1、OAS2、OASL的mRNA表达。结果SLE活动组与正常对照组相比OAS1、OAS2、OASL的表达均明显增高(P<0.01),感染组3者表达均较SLE活动组为低(P<0.01)。SLE合并感染组与正常对照组相比OAS1的表达仍增高(P=0.000)、OAS2的表达增高不明显(P=0.124)、OASL的表达则明显降低(P=0.01)。Logistic回归显示OASL表达减低构成SLE合并感染的预测因子(P<0.01)。结论初步提示OAS异构体表达格局,特别是OASL,可能成为SLE活动和感染鉴别诊断的参考指标。     

寡腺苷酸合成酶家族基因的定表达与系统性红斑狼疮的临床特征

目的探讨系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中寡腺苷酸合成酶(OAS)1、OAS2和OASLmRNA的实时定量表达水平与SLE疾病特异性和病情活动度的相关性。方法收集144例SLE患者、27例非SLE患者与59名正常对照人群的临床资料,取外周血抽取总RNA并反转录成cDNA,运用sybrgreendyeⅠ实时定量聚合酶链反应(PCR)法在ABIPRISM7900H基因测序仪上检测患者组和对照组的OAS1、OAS2的OASLmRNA表达水平的差异,并与病情活动度进行分组比较,分析其意义。结果①SLE患者的总体OAS1mRNA定量表达水平显著高于非SLE对照组和正常对照组,差异有非常显著性(P=0郾016,P=0郾000);OAS2mRNAP定量表达水平高于正常对照组(P=0郾015);而OASLmRNA定量表达水平与非SLE对照组和正常组之间差异无显著性(P=0郾698,P=0郾396)。②SLE活动组的OAS1和OASLmRNA定量表达水平均显著高于SLE非活动组,差异有非常显著性(P=0郾011,P=0郾001)。但SLE活动组的OAS2表达水平与SLE非活动组之间差异无显著性(P=0郾109)。③SLE患者组的OAS2和OASLmRNA定量表达水平与SLEDAI积分呈正相关性(P=0郾001,P=0郾006)。④SLE患者的肾、肺、脑、血液等器官的有无受累与OAS1、OAS2和OASLmRNA表达水平无相关性。⑤SLE患者组和正常对照组组内的OAS1、OAS2和OASL表达水平     

系统性红斑狼疮外周血细胞干扰素诱导蛋白IFI16 IFI27及IFI30表达研究

目的 探讨干扰素诱导蛋白(IFI 16、IFI 27、IFI 30)mRNA表达与系统性红斑狼疮(SLE)的关系.方法 以前期构建SLE患者外周血单个核细胞(PBMCs)基因表达系列分析(SAGE)文库为基础,分析该文库中出现的3个干扰素相关基因标签(tag),并采用反转录-聚合酶链反应(RT-PCR)方法,临床检测IFI16、IFI27、IFI30在43例SLE患者和25名正常对照人群PBMCs中的mRNA表达水平.结果 分别约有88%、98%、58%的SLE个体其IFI16、IFI27、IFI30 mRNA表达增加,高于正常对照组总体均数95%可信区间的上限.与正常对照组比较,SLE活动组和缓解IFI16、IFI27、IFI30的表达水平均增加(P值均＜0.01).活动组与缓解组间比较,活动组SLE患者高表达IFI16和IFI27(P=0.003,P=0.001),IFI30的表达差异无统计学意义(P=0.227). SLE患者IFI16、IFI27、IFI30间的表达水平互不相关.IFI16、IFI27表达与狼疮疾病活动指数(SLEDAI)呈正相关,相关系数分别为0.557、0.414,P直分别为0.000,0.006.IFI30表达与SLEDAI无相关(r=-0.102,P=0.514).结论 SLE患者IFI16、IFI27、IFI30表达增高,揭示其在SLE发病中的重要作用.     

干扰素调节因子5在系统性红斑狼疮的表达及临床相关性研究

目的 比较干扰素调节因子5(IRF5)mRNA在系统性红斑狼疮(SLE)患者和健康对照组的表达水平,分析IRF5表达与SLE疾病活动性及自身抗体和临床症状的相关性.方法 Ficoll密度梯度离心法分离SLE患者及健康对照外周血单个核细胞(PBMC),Trizol法分离提取总RNA,反转录mRNA为eDNA;实时定量聚合酶链反应(PCR)法测定SLE患者和正常对照组IRF5表达量;并分析SLE患者IRF5表达量与疾病活动性及临床症状的相关性.结果 SLE组IRF5表达量(2.1+2.2)高于正常对照组(1.5±1.2),但差异无统计学意义(P=0.161);SLE患者IRF5表达量与其疾病活动指数(SLEDAI)显著相关(r=0.616,P<0.01);SLE患者中抗dsDNA抗体阳性组IRF5表达量(3.2±2.8)明显高于抗体阴性组(1.3±1.6).差异有统计学意义(P=0.018);SLE患者有发热、神经精神症状者IRF5表达量明显高于无此类临床症状者.结论 IRF5在SLE患者中表达偏高,且与SLEDAI显著相关,其可能通过调节下游基因的转录表达诱一导免疫失调,并由此参与SLE的发病过程.     

狼疮肾炎患者Ⅰ型干扰素诱导基因表达的研究

目的 了解Ⅰ型干扰素(IFN)诱导基因IFIT4、IFI44、Ly6e、OAS1、OAS2和OAS3在狼疮肾炎(LN)患者的表达,并分析IFN积分与临床特征之间的相关性.方法 ①同步收集12例弥漫增生型LN患者.肾组织和外周血,抽提总RNA并反转录为cDNA,以实时荧光定量聚合酶链反应(PCR)方法检测Ⅰ型IFN诱导基因IFIT4、IFI44、Ly6e、OAS1、OAS2和OAS3的表达水平.②收集119例LN患者的临床资料,计算每例患者的IFN积分,并与系统性红斑狼疮疾病活动指数(SLEDAI)积分、抗dsDNA抗体、补体、蛋白尿水平等指标进行相关性检验.结果 ①弥漫增生型LN患者肾组织高表达OAS2[(1.46±1.15)与(0.60±0.59)]和OAS3[(2.16±1.63)与(0.93±0.95)],P<0.05;IFIT4[(1.68±1.76)与(0.96±0.71)]、IFI44[(3.23±2.67)与(1.65±1.95)]、Ly6e[(3.54±2.09)与(2.34±2.57)]、OAS1[(2.52±1.86)与(1.34±0.28)]的表达量高于对照组1个循环左右.狼疮患者外周血高表达IFIT4[(11.2±7.7)与(1.56±2.71)]、IFI44[(9.5±6.1)与(2.6±4.4)J、Ly6e[(17.5±13.7)与(7.1±10.3)]、OAS1[(8.68±6.36)与(1.15±1.89)]、OAS2[(10.41±7.71)与(0.66±0.82)]、OAS3[(8.75±4.34)与(0.89±1.34)],P<0.05.Ⅰ型IFN诱导基因IFIT4、IFI44、Ly6e、OAS1、OAS2和OAS3在肾脏组织和外周血的表达呈同步增高趋势.②根据119例患者IFIT4、IFI44、Ly6e、OAS1、OAS2和OAS3的表达计算IFN积分,发现IFN积分与SLEDAI、抗dsDNA抗体呈正相关(r分别是0.216和0.394,P<0.05),与血清补体水平呈负相关(r=0.315,P<0.01).结论 Ⅰ型IFN诱导基因IFIT4、IFI44、Ly6e、OAS1、OAS2和OAS3外周血表达的水平在一定程度上反映LN肾损害情况,IFN积分对监测LN患者的疾病活动有一定的特异性.     

系统性红斑狼疮患者磷脂酶C-γ1与磷脂酶C-γ2表达的研究

目的 探讨系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMCs)中磷脂酶C-γ1、磷脂酶C -γ2的表达水平及其与SLE疾病活动性的相关性.方法 运用半定量聚合酶链反应(RT-PCR)法,检测30例SLE患者与25名健康对照人的磷脂酶C-γ1与磷脂酶C-γ2 mRNA表达水平,并采用Pearson相关性检验分析它们与补体C3、C4、抗双链DNA (dsDNA)抗体及SLE疾病活动指数(SLEDAI)积分的相关性.结果 ①SLE组与健康对照组相比,磷脂酶C-γ1与磷脂酶C-γ2 mRNA表达显著增高(分别为P＜0.01);②磷脂酶C-γ1与磷脂酶C-γ2 mRNA表达水平之间呈正相关(r=0.726,P＜0.01);③SLE组磷脂酶C-γ1 mRNA表达水平与补体C3、C4、抗dsDNA抗体均无相关性,而与SLEDAI积分呈正相关(r=0.002,P＜0.05).磷脂酶C-γ2与补体C3、C4呈显著负相关(r=-0.914,P＜0.01;r=-0.829,P＜0.05),与抗dsDNA抗体正相关(r=0.171,P＜0.05),与SLEDAI积分相关性不明显.结论 本研究发现SLE患者磷脂酶C-γ1与磷脂酶C-γ2的表达水平在SLE患者中增高,磷脂酶C-γ1与SLEDAI积分呈正相关,磷脂酶C-γ2与补体C3、C4呈负相关,与抗dsDNA抗体正相关,可能对SLE患者的诊断和病情评估有较大价值.     

Association of increased interferon-inducible gene expression with disease activity and lupus nephritis in patients with systemic lupus erythematosus

OBJECTIVE: To study 5 type I interferon (IFN)-inducible genes (LY6E, OAS1, OASL, MX1, and ISG15) in patients with systemic lupus erythematosus (SLE) and to correlate expression levels with disease activity and/or clinical manifestations. METHODS: Peripheral blood cells were obtained from 48 SLE patients, 48 normal controls, and 22 rheumatic disease controls, and total RNA was extracted and reverse transcribed into complementary DNA. Gene expression levels were measured by real-time polymerase chain reaction, standardized to a housekeeping gene, and summed to an IFN score. Disease activity was determined by the Safety of Estrogens in Lupus Erythematosus: National Assessment-Systemic Lupus Erythematosus Disease Activity Index (SELENA-SLEDAI) composite. RESULTS: Each gene was highly expressed in SLE patients compared with normal controls (P < or = 0.0003) or disease controls (P < or = 0.0008 except for MX1). IFN scores were positively associated with the SELENA-SLEDAI instrument score (P = 0.001), the SELENA-SLEDAI flare score (P = 0.03), and the physician's global assessment score (P = 0.005). Compared with patients without nephritis, lupus nephritis patients had higher IFN scores (overall P < 0.0001), especially during active renal disease. IFN scores were weakly associated with neurologic manifestations. Elevated IFN scores were positively associated with the current presence of anti-double-stranded DNA (anti-dsDNA) antibodies (P = 0.007) or hypocomplementemia (P = 0.007). LY6E expression levels distinguished active from inactive lupus nephritis (P = 0.02) and were positively associated with proteinuria (P = 0.009). CONCLUSION: The 5 IFN-inducible genes were highly expressed in SLE patients, and increased levels were correlated with disease activity defined by several methods. IFN scores, or LY6E levels, were elevated in lupus nephritis patients, especially during active renal disease, and in patients with anti-dsDNA antibody positivity and hypocomplementemia. IFN scores, or LY6E levels, may be useful as a biomarker for lupus nephritis therapy.     

Association of increased interferon‐inducible gene expression with disease activity and lupus nephritis in patients with systemic lupus erythematosus

Objective

To study 5 type I interferon (IFN)–inducible genes (LY6E, OAS1, OASL, MX1, and ISG15) in patients with systemic lupus erythematosus (SLE) and to correlate expression levels with disease activity and/or clinical manifestations.

Methods

Peripheral blood cells were obtained from 48 SLE patients, 48 normal controls, and 22 rheumatic disease controls, and total RNA was extracted and reverse transcribed into complementary DNA. Gene expression levels were measured by real‐time polymerase chain reaction, standardized to a housekeeping gene, and summed to an IFN score. Disease activity was determined by the Safety of Estrogens in Lupus Erythematosus: National Assessment–Systemic Lupus Erythematosus Disease Activity Index (SELENA‐SLEDAI) composite.

Results

Each gene was highly expressed in SLE patients compared with normal controls (P ≤ 0.0003) or disease controls (P ≤ 0.0008 except for MX1). IFN scores were positively associated with the SELENA‐SLEDAI instrument score (P = 0.001), the SELENA‐SLEDAI flare score (P = 0.03), and the physician's global assessment score (P = 0.005). Compared with patients without nephritis, lupus nephritis patients had higher IFN scores (overall P < 0.0001), especially during active renal disease. IFN scores were weakly associated with neurologic manifestations. Elevated IFN scores were positively associated with the current presence of anti–double‐stranded DNA (anti‐dsDNA) antibodies (P = 0.007) or hypocomplementemia (P = 0.007). LY6E expression levels distinguished active from inactive lupus nephritis (P = 0.02) and were positively associated with proteinuria (P = 0.009).

Conclusion

The 5 IFN‐inducible genes were highly expressed in SLE patients, and increased levels were correlated with disease activity defined by several methods. IFN scores, or LY6E levels, were elevated in lupus nephritis patients, especially during active renal disease, and in patients with anti‐dsDNA antibody positivity and hypocomplementemia. IFN scores, or LY6E levels, may be useful as a biomarker for lupus nephritis therapy.

     

Could 2′5′-oligoadenylate synthetase isoforms be biomarkers to differentiate between disease flare and infection in lupus patients? A pilot study

2′5′-Oligoadenylate synthetase (OAS) was shown to be related to systemic lupus erythematosus (SLE) 20 years ago, and was rediscovered to be involved in type I interferon pathway in SLE by several microarray gene expression studies recently. The goal of this study was to investigate OAS isoform expressions in lupus patients, to evaluate whether they could become biomarkers to differentiate between disease flare and infection. Fifty-four SLE patients presented with fever or systemic inflammatory syndrome, or both, were enrolled. Gene expressions of OAS1, OAS2, and OASL were studied by using real time PCR in active SLE (SLEDAI ≥9, n=29) and in those complicated with infections (n=25). The latter group was composed of 19 patients with invasive bacterial infections, and six patients with viral infections. C reactive protein (CRP) and other clinical parameters were also measured. Twenty-nine healthy individuals made up a normal control group. The mRNA expressions of OAS1, OAS2, and OASL were higher in patients with lupus flares than those with infections (p<0.03), or normal controls (p<0.001). SLE complicated with infections have higher OAS1 expression level (P=0.002), lower OASL (P=0.004), and equivalent OAS2 (P=0.135), when compared with those of normal controls. OASL expression level was negatively correlated with infection in lupus by logistic regression analysis (p=0.008). Area under the receiver operating characteristic curve for the prediction of infection was 0.92 (p<0.0001) for OASL, and 0.77 (p=0.007) for CRP. Therefore, our preliminary data suggest that the pattern of OAS isoform expressions, OASL in particular, may provide useful information in differentiating disease flares from certain infections in SLE.     

Higher activation of the interferon-gamma signaling pathway in systemic lupus erythematosus patients with a high type I IFN score: relation to disease activity

Increased IFN-γ levels have been associated with systemic lupus erythematosus (SLE). However, the relationships among IFN-γ, type I interferons (IFNs) and clinical features have not been extensively studied. Peripheral blood samples from 44 SLE patients and 36 healthy donors (HDs) were collected. Quantitative real-time PCR was used to assess the mRNA expression of IFNG, type II IFN-inducible genes (IRF1, GBP1, CXCL9, CXCL10, and SERPING1, which are used for the type II IFN score), type I IFN-inducible genes (IRF7, MX1, ISG15, and ISG20, which are used for the type I IFN score), TBX21, and EOMES in peripheral blood mononuclear cells. Flow cytometry was used to measure the IFN-γ levels in lymphocytes. The mRNA levels of type II IFN-inducible genes, IFNG, TBX21, and EOMES were significantly higher in SLE patients than those in HDs. Similarly, the percentages of IFN-γ-producing cells in lymphocytes and their subsets in SLE patients were significantly increased. Linear regression indicated that IFNG expression levels and type II IFN scores were positively correlated with anti-double-stranded DNA autoantibody levels and Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) scores. Compared with patients with low type I IFN scores, patients with high type I IFN scores showed increased type II IFN scores and SLEDAI scores. Type II IFN scores were positively associated with type I IFN scores. The IFN-γ signaling pathway is activated in SLE patients and may be considered an index of disease activity. IFN-γ, together with type I IFNs, promotes the pathogenesis of SLE.     

Ⅰ型干扰素系统在自身免疫性炎性肌病大鼠肌肉和肺组织的表达

目的 通过观察自身免疫性炎性肌病(EAM)大鼠肌肉和肺组织Ⅰ型IFN系统的表达,初步探讨EAM合并肺间质病变的病理机制。方法 建立EAM大鼠模型,分为健康对照组、EAM模型对照组、EAM模型组,测3组大鼠血清中肌酸激酶(CK)水平,HE染色分析模型大鼠肌肉和肺脏病变程度,实时定量PCR测Ⅰ型IFN系统在肌肉和肺组织中的表达。结果 与健康对照组、EAM模型对照组比,EAM模型组血清CK[(209.17±91.95) IU/L]、肌肉病理评分[(2.17±0.76)分]和肺脏组织病理评分[(1.75 ±0.41)分]、肌肉组织中Ⅰ型IFN系统mRNA表达水平明显升高,差异有统计学意义(P值均小于0.05)。与EAM模型对照组比,EAM模型组肺组织IFNα、IFNβ、干扰素α受体1(IFNαR1)、信号传导蛋白和转录激活物1(STAT1)、黏病毒抑制蛋白1(MX1) mRNA表达水平升高,差异有统计学意义(P值均小于0.05),而干扰素诱导蛋白1(IFIT1)、干扰素诱导基因15(ISG15)表达水平差异无统计学意义。EAM模型组大鼠肌肉组织中IFNα、IFNβ、IFNαR1、STAT1、MX1、IFIT1、ISG15 mRNA表达与血清CK水平和肌肉病理评分呈正相关。EAM模型组大鼠肺组织中IFNα、IFNβ、IFNαR1、STAT1、MX1 mRNA表达与肺脏病理评分呈正相关。结论 EAM大鼠肌肉和肺组织均有Ⅰ型IFN系统表达,Ⅰ型IFN可能参与了EAM大鼠靶器官的病理损害。