女性系统性红斑狼疮患者外周血Tim-1表达水平及其与疾病活动的相关性

目的:检测T细胞免疫球蛋白粘蛋白分子-1在女性系统性红斑狼疮(SLE)患者外周血的表达情况,并探讨与SLE疾病活动的相关性.方法:采用酶联免疫吸附测定(ELISA)方法检测SLE治疗前后和健康对照组外周血中T细胞免疫球蛋白粘蛋白分子-1的表达量,SLE活动性以系统性红斑狼疮病变活动指数(SLEDAI)表示,并进行相关性分析以明确T细胞免疫球蛋白粘蛋白分子-1与系统性红斑狼疮患者疾病活动性的关系.结果:治疗前SLE患者外周血中T细胞免疫球蛋白粘蛋白分子-1的表达水平明显高于对照组和治疗后SLE患者外周血中的表达水平,有统计学差异(P0.05),且其表达水平与SLEDAI评分呈正相关(r=0.639,P0.01).结论:T细胞免疫球蛋白粘蛋白分子-1的表达水平与SLE的发病有关,且可反映SLE患者的疾病活动状态.     

白介素-38在活动期系统性红斑狼疮患者中的表达及与IKKβ激酶的关系

目的:研究活动期系统性红斑狼疮(SLE)患者外周血中白介素-38(IL-38)和IKKβ激酶的表达水平,以探讨IL-38与IKKβ的关系.方法:采用实时荧光定量聚合酶链反应检测IL-38、IKK基因表达水平;酶联免疫吸附试验(ELISA)检测血清IL-38、肿瘤坏死因子α(TNF-α)蛋白表达水平;采用t检验或Mann-Whitney秩和检验.结果:(1)活动期SLE患者IL-38 mRNA表达水平(0.36±0.09)显著低于正常对照组(1.00±0.17),差异有统计学意义(P0.01);活动期SLE患者IL-38蛋白表达水平(5.86±2.76)pg/mL显著低于正常对照组(18.48±1.35)pg/mL,差异有统计学意义(P0.05);(2)活动期SLE患者TNF-α表达水平(10.78±1.25)pg/mL显著高于正常对照组(4.34±0.69)pg/mL,差异有统计学意义(P0.05);(3)活动期SLE患者IKKβmRNA表达水平(6.01±1.51)显著高于正常对照组(1.16±0.14),差异有统计学意义(P0.05);(4)活动期SLE患者IL-38与TNF-α蛋白表达水平呈负相关,差异有统计学意义(P0.05);(5)活动期SLE患者IL-38与IKKβmRNA表达水平呈负相关,差异有统计学意义(P0.05).结论:活动期SLE患者IL-38基因及蛋白表达水平下降,并且与TNF-α和IKKβ水平呈负相关,推测在SLE患者体内IL-38水平下降,促进TNF-α和IKKβ的高表达,从而活化核因子-κB(NF-κB)信号通路,参与了SLE的发病.     

解毒活血滋阴方对系统性红斑狼疮患者血清Blys、IL-10的影响

本文旨在探讨解毒活血滋阴方对系统性红斑狼疮(SLE)患者血清B淋巴细胞刺激因子(Blys)、IL-10的影响。将65例SLE患者随机分为2组:西医组(32例)采用强的松、甲氨蝶呤、羟氯喹治疗;中西医结合组(33例)采用解毒活血滋阴方联合强的松、甲氨蝶呤、羟氯喹治疗,均治疗3个月。运用酶联免疫吸附试验(ELISA)法检测血清Blys、IL-10的含量。结果显示,与治疗前相比,西医组、中西医结合组血清Blys、IL-10含量均明显降低(P<0·01),且中西医结合组Blys、IL-10的改善更加明显(P<0.05,P<0.01)。同时,西医组、中西医结合组均能明显降低SLE患者系统性红斑狼疮狼疮活动指数(SLEDAI)积分(P<0.01),且中西医结合组SLEDAI积分的改善更加明显(P<0.05)。结论:解毒活血滋阴方能明显降低SLE患者血清Blys、IL-10的含量,这可能是其作用机制之一。     

系统性红斑狼疮患者肿瘤坏死因子可溶性受体水平的测定

目的　探讨血清中可溶性肿瘤坏死因子受体 (sTNFR)在系统性红斑狼疮 (SLE)中的变化规律 .方法ELISA法测定血清sTNFR的蛋白水平 .结果　SLE中sTNFR蛋白水平比正常对照显著增高 (P <0 .0 0 1) .活动期比非活动期增高亦显著 (P <0 .0 1) .结论　sTNFR是SLE疾病表现强度的主要因素 ,sTNFR可作为SLE活动期判定的指标 .     

干扰素调节因子的研究进展

指出了干扰素调节因子(IRFs)是一类在干扰素(IFN)信号通路中起重要调控作用的多功能转录因子,目前已经发现10个IRFs成员,它们在IFN的诱导、病毒防御、免疫调节、细胞分化、细胞生长与凋亡和许多疾病的调节中具有重要作用.对IRFs的结构、IRFs在免疫系统、细胞分化、细胞凋亡和相关疾病如肿瘤、系统性红斑狼疮中的功能作了综述.     

SLE患者淋巴细胞凋亡及血清新蝶呤、γ-干扰素水平研究

探讨SLE患者体内淋巴细胞凋亡率、巨噬细胞功能相关的细胞因子(新蝶呤、γ-干扰素)水平及两者间相互关系,并同时与抗ds-DNA抗体滴度、疾病活动性评分(SLAM评分)进行相关性分析.应用Annexin V凋亡检测试剂盒及流式细胞仪检测淋巴细胞凋亡率;采用ELISA法检测血清γ-干扰素、新蝶呤水平.结果发现:(1)SLE活动期患者外周血淋巴细胞凋亡率((13.07±7.39),n=30)明显高于非活动期患者((4.08±3.55),n=8,P<0.001)及正常人((5.13±3.37),n=11,P<0.001).(2)抗ds-DNA抗体阳性组患者淋巴细胞凋亡率((12.98±9.25),n=20)与阴性组((9.35±4.76),n=18)相比无差异(P=0.14).淋巴细胞凋亡率与抗ds-DNA抗体滴度无相关性(r=0.112,P=0.77).(3)SLE患者血清新蝶呤水平((1.39±1.10)μg/L,n=20)极显著高于正常人((0.36±0.19)μg/L,n=20,P<0.01).(4)SLE活动期患者血清γ-干扰素水平((58.97±34.52)ng/L,n=15)显著高于正常人((28.06±2.35)ng/L,n=16,P<0.05).(5)SLE患者淋巴细胞凋亡率与血清新蝶呤水平呈正相关(r=0.446,P<0.05,n=22),与SLAM评分呈正相关(r=0.533,P<0.001,n=38),血清新蝶呤水平与SLAM评分亦呈正相关(r=0.485,P<0.05,n=22).未发现SLE患者淋巴细胞凋亡与抗ds-DNA抗体产生相关,未发现明显升高的细胞凋亡率与血清新蝶呤     

精细化护理干预在系统性红斑狼患者护理中应用的价值

探究精细化护理干预在系统性红斑狼疮患者护理中应用的价值。选取兰州大学第二医院收治的系统性红斑狼疮患者174例,采用随机数表法分为观察组和对照组,对照组给予常规护理,观察组实施精细化护理,对比两组患者心理状况、治疗依从性。观察组患者护理后SCL-90各因子评分均显著低于对照组且组间差异存在统计学意义(P0.05);观察组患者治疗依从性89.66%明显高于对照组66.67%(P0.05)。系统性红斑狼患者应用精细化护理干预可改善负性情绪,提高治疗依从性。     

SLE患者外周血γδT细胞系的建立及表型和功能的研究

目的:建立体外扩增系统性红斑狼疮(SLE)患者外周血γδT细胞的方法,并初步分析其表型和功能,探索γδT细胞在SLE发病中的作用。方法:采用单克隆抗体固相法,对15例SLE患者和8例正常人外周血γδT细胞进行体外扩增建系,以流式细胞仪检测γδT细胞表型,并用MTT法观察γδT细胞对Daud i细胞的细胞毒作用。结果:建立了SLE患者外周血γδT细胞系,其平均纯度为(58.1±11.2)%,较正常对照组(80.3±9.2)%偏低(P<0.05);其细胞表型为:Vδ1(34.4±24.5)%、Vδ2(61.9±28.6)%、Vδ3(16.1±10.6)%、Vγ9(76.4±11.8)%,其中Vδ2表达较对照组降低,而Vδ1和Vδ3表达增加(P均<0.05);其细胞毒作用在二组间无明显差别。结论:SLE患者外周血γδT细胞的Vδ、Vγ基因的取用表达存在差异,提示其在SLE的发病中可能起一定作用。     

系统性红斑狼疮TH2型细胞因子受体基因的表达

目的 分析SLE患者外周血单一核细胞TH2型细胞因子mRNA的表达状况.方法 分离SLE外周血单一核细胞1×107/mL的浓度,进行总RNA提取,然后用1.5%的琼脂糖电泳,应用凝胶成像系统对电泳条带进行扫描.结果 SLE患者IL-4mRNA,IL-6mRNA和IL-10mRNA水平相比对照组显著升高.结论 TH2型细胞因子受体基因的表达在SLE发病机制中有明显的异常,其与相应蛋白表达的关系还有待进一步探讨.     

用多克隆抗体检测干扰素调节因子7的表达

干扰素调节因子7(IRF-7)是调节Ⅰ型干扰素依赖型先天免疫反应的关键因子,在真核细胞防御反应中发挥重要作用。本文在大肠杆菌中表达了重组IRF-7,利用亲和层析的方法进行了纯化,并制备了鼠抗IRF-7的多克隆抗体。所得到的抗血清能够在稀释27 000倍后成功用于免疫印迹检测。该抗血清不但能识别来源于大肠杆菌的抗原,还能检测真核细胞内转染后表达的IRF-7。使用该抗体证实,转染293T细胞后表达的IRF-7主要分布在细胞质内。所制备的抗血清可用于IRF-7的表达和功能研究。     

Critical role of TRAF3 in the Toll-like receptor-dependent and -independent antiviral response

Type I interferon (IFN) production is a critical component of the innate defence against viral infections. Viral products induce strong type I IFN responses through the activation of Toll-like receptors (TLRs) and intracellular cytoplasmic receptors such as protein kinase R (PKR). Here we demonstrate that cells lacking TRAF3, a member of the TNF receptor-associated factor family, are defective in type I IFN responses activated by several different TLRs. Furthermore, we show that TRAF3 associates with the TLR adaptors TRIF and IRAK1, as well as downstream IRF3/7 kinases TBK1 and IKK-epsilon, suggesting that TRAF3 serves as a critical link between TLR adaptors and downstream regulatory kinases important for IRF activation. In addition to TLR stimulation, we also show that TRAF3-deficient fibroblasts are defective in their type I IFN response to direct infection with vesicular stomatitis virus, indicating that TRAF3 is also an important component of TLR-independent viral recognition pathways. Our data demonstrate that TRAF3 is a major regulator of type I IFN production and the innate antiviral response.     

Interferon-dependent induction of mRNA activity for (2'-5')oligo-isoadenylate synthetase

At least three different enzymes involved in the regulation of protein synthesis are induced in a variety of cells by interferon (IFN). Sensitive assays for these enzymes have been developed and used to establish the specificity, dose dependence and time course of their induction by IFN. One of these enzymes, the oligo-isoadenylate synthetase E, whose product (2'-5')pppApApA activates the latent ribonuclease F, is increased over 50-fold after IFN treatment. We describe here the assay for an mRNA from IFN-treated mouse L cells, that produces oligo-isoadenylate synthetase activity when injected into Xenopus oocytes. This mRNA is found in the cells only after exposure to IFN. The mRNA increases in mouse L cells with the same time course as the enzyme activity itself. In particular, there is a 3-h lag period between IFN addition and the onset of enzyme and mRNA accumulation. Using anti-IFN antibodies, we show that during this lag period the continued interaction of IFN with the cells is necessary for the full induction of the oligo-isoadenylate synthetase.     

2'-O methylation of the viral mRNA cap evades host restriction by IFIT family members

Cellular messenger RNA (mRNA) of higher eukaryotes and many viral RNAs are methylated at the N-7 and 2'-O positions of the 5' guanosine cap by specific nuclear and cytoplasmic methyltransferases (MTases), respectively. Whereas N-7 methylation is essential for RNA translation and stability, the function of 2'-O methylation has remained uncertain since its discovery 35 years ago. Here we show that a West Nile virus (WNV) mutant (E218A) that lacks 2'-O MTase activity was attenuated in wild-type primary cells and mice but was pathogenic in the absence of type I interferon (IFN) signalling. 2'-O methylation of viral RNA did not affect IFN induction in WNV-infected fibroblasts but instead modulated the antiviral effects of IFN-induced proteins with tetratricopeptide repeats (IFIT), which are interferon-stimulated genes (ISGs) implicated in regulation of protein translation. Poxvirus and coronavirus mutants that lacked 2'-O MTase activity similarly showed enhanced sensitivity to the antiviral actions of IFN and, specifically, IFIT proteins. Our results demonstrate that the 2'-O methylation of the 5' cap of viral RNA functions to subvert innate host antiviral responses through escape of IFIT-mediated suppression, and suggest an evolutionary explanation for 2'-O methylation of cellular mRNA: to distinguish self from non-self RNA. Differential methylation of cytoplasmic RNA probably serves as an example for pattern recognition and restriction of propagation of foreign viral RNA in host cells.     

Heterogeneity of poly(I) x poly(C)-induced human fibroblast interferon mRNA species

Three classes of human interferons (IFNs) have been defined on the basis of their immunological properties: the 'Le' or 'alpha' IFN, mainly derived from leukocyte or lymphoblastoid cells; the 'F' or 'beta' IFN, mainly derived from fibroblast cultures; and the 'T', 'immune' or 'gamma' IFN, mainly derived from mitogen- or antigen-stimulated lymphoid cells. Whereas several individual species of Le IFN have been purified to homogeneity, it is generally considered that F IFN represents a single protein. Thus current efforts to clone human fibroblast IFN mRNA sequences are based on the observation that F IFN mRNA sediments in sucrose gradients as a single RNA species of size corresponding to 12-14 S (refs 7-10). We show here, using gel electrohporesis of mRNA, that two populations of translationally active human fibroblast IFN mRNA molecules exist--an abundant '14 S' species and a scarce '11 S' species. Microinjection of either species of mRNA into Xenopus oocytes leads to the synthesis of biologically active F-type human IFN. These data agree with and complement recent RNA hybridization studies of Weissenbach et al.     

Interferon-neutralizing antibodies in a patient treated with human fibroblast interferon

During recent years clinical trials have shown that human leukocyte interferon (HuIFN-alpha) may be useful in the treatment of cancer, but very little has been done concerning the possible use of human fibroblast interferon (HuIFN-beta). Treuner et al. recently reported the successful treatment of a nasopharyngeal carcinoma with HuIFN-beta: in the course of IFN-therapy a HuIFN-beta neutralizing activity appeared in the serum of this patient. We report here that such activity is due to IgG antibodies--this study is the first to present evidence for antigenicity of IFN in a homologous system.