白细胞介素10免疫调节功能的研究进展

白细胞介素10（IL-10）作为一种多效价的细胞因子,最初认为其主要由Th2细胞产生,随后的研究发现Th1、Th17、调节性T细胞（Treg）、Th9以及CD8＋T细胞和B细胞均可以产生IL-10 另外固有免疫细胞以及角质形成细胞、上皮细胞和肿瘤细胞等也产生IL-10.IL-10通过与细胞表面IL-IOR1和IL-IOR2相互作用并主要激活JAK.STAT通路发挥生物学作用.IL-10可以抑制抗原提呈细胞的活化,共刺激分子的表达以及提成抗原的能力,降低促炎性细胞因子的合成,参与免疫球蛋白（Ig）的类别转换以及介导肿瘤细胞免疫逃逸等的发生.IL-10在炎症,多种自身免疫病以及肿瘤,移植排斥反应等多种疾病的发生和发展过程中都有重要的免疫调节作用.     

白细胞介素10的细胞生物学效应及其信号转导

IL 10是一种由多种细胞生成的具有多种生物学活性的细胞因子 ,调节T细胞、B细胞、肥大细胞及造血细胞的分化发育 ,在炎症免疫反应、肿瘤、病毒感染、造血系统等多方面发挥重要作用。在不同的细胞中IL 10作用不同 ,其信号转导通路也存在差异。本文就IL 10在不同细胞中的生物学效应及其信号传导通路综述如下     

106 白细胞介素10的细胞生物学效应及其信号转导

IL-10是一种由多种细胞生成的具有多种生物学活性的细胞因子,调节T细胞、B细胞、肥大细胞及造血细胞的分化发育,在炎症免疫反应、肿瘤、病毒感染、造血系统等多方面发挥重要作用.在不同的细胞中IL-10作用不同,其信号转导通路也存在差异.本文就IL-10在不同细胞中的生物学效应及其信号传导通路综述如下.     

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。     

地塞米松双向调节小鼠腹腔巨噬细胞白细胞介素-10的表达

IL 10是从TH2细胞分离到的 ,因能抑制TH1细胞合成和释放细胞因子 ,故又名细胞因子合成抑制因子。IL 10主要针对T细胞、NK细胞、单核巨噬细胞起抑制作用 ,是影响体内TH1和TH2平衡的关键性细胞因子之一。因此 ,对免疫炎症性疾病的发生和发展具有重要的调节作用。我们研究了地塞米松(Dex)对小鼠腹腔巨噬细胞 (MΦ)IL 10表达的影响。材料和方法试剂 :Dex购自上海兴宜制药有限公司 ,脂多糖(LPS)及糖皮质激素受体拮抗剂RU4 86购自Sigma ,RPMI 16 40培养基及总RNA抽提用TRIZOL试剂购自GIBCO BRL ,逆转录试剂盒购自MBIFermenta…     

内毒素对C57BL/6纯种小鼠腹腔巨噬细胞糖皮质激素受体的调节作用

本文用5—3周龄雌性C57BL/6纯种小鼠(上海医药工业研究院提供),颈椎脱臼处死,在无菌条件下迅速分离和暴露腹膜,将10ml Hank's液注入腹腔,按摩腹壁150次,抽出冲洗液,离心(1200rpm,10min),弃上     

肺炎衣原体对C57BL/6J小鼠腹膜巨噬细胞泡沫化的影响

目的:探讨肺炎衣原体在C57BL/6J小鼠腹膜巨噬细胞泡沫化过程中的作用。方法:将肺炎衣原体与C57BL/6J小鼠腹膜巨噬细胞培养72h后用油红O方法染色观察细胞的形态变化并测定细胞内胆固醇的含量。结果:肺炎衣原体能使C57BL/6J小鼠腹膜巨噬细胞胞浆内脂质颗粒增多,胆固醇酯占总胆固醇的比例增加。结论:肺炎衣原体能促进C57BL/6J小鼠腹膜巨噬细胞泡沫化。     

SLE患者外周血淋巴细胞凋亡异常及血清中IL-10的影响

目的：研究SLE患者外周血单个核细胞(PBMCs)中淋巴细胞亚群的凋亡特点。方法：采用三色荧光流式细胞术检测CD3、CD4、CD8、CD19细胞亚群的早期凋亡；ELISA法检测PBMCs培养上清中的sFas、sFasL；实时荧光半定量RT-PCR方法检测细胞内FasmRNA的表达水平。结果：SLE患者PBMCs中CD3^ 细胞凋亡明显增多；CD4^ 和CD8^ 细胞亚群凋亡均有明显增加，但以CD4^ 细胞凋亡增多更为明显；相应地，其PBMCs中CD4^ 、CD8^ 细胞百分率下降，CD4／CD8比值降低。SLE患者血清可诱导CD3^ 、CD4^ 、CD8^ T细胞亚群凋亡增多，sFas、sFasL水平增高及Fas mRNA表达增加；抗-IL-10抗体则可中和SLE血清的上述作用。结论：SLE患者体内T细胞凋亡增多，以CD4^ T细胞凋亡增加更为显著，导致CD4／CD8比值下降；SLE血清中高水平的IL-10可能通过诱导Fas、FasL表达增高而促进T细胞凋亡的发生。     

激活FXR抑制MRL/lpr狼疮小鼠肝损害的研究

 目的:观察法尼酯衍生物X受体（FXR）激活是否减轻MRL/lpr狼疮小鼠肝损害。方法:检测FXR在MRL/lpr狼疮小鼠及正常BALB/c小鼠肝脏的表达;观察BALB/c小鼠和鹅去氧胆酸（CDCA）激活FXR的MRL/lpr小鼠再以伴刀豆球蛋白A(ConA)诱导肝脏炎症反应后的肝脏酶学、炎症因子及病理学的变化。结果:FXR在MRL/lpr狼疮小鼠肝脏中低表达;ConA能在MRL/lpr小鼠诱导出较对照BALB/c小鼠更严重的肝损害;激活FXR减轻MRL/lpr狼疮小鼠ConA诱导的肝脏炎症损伤,并减少一系列炎症因子的释放。结论: CDCA激活FXR 后,能减轻狼疮小鼠肝脏损害。FXR可能是系统性红斑狼疮肝损害的一个保护性因素,激活FXR可能成为狼疮肝损害的一个治疗途径。     

系统性红斑狼疮患者血清IL-10和IL-18的检测及意义

目的探讨系统性红斑狼疮（SLE）患者血清IL-10和IL-18的表达及其与疾病活动的关系。方法应用ELISA法检测104例SLE患者和100例健康体检者血清IL-10和IL-18的水平,其中SLE患者根据疾病活动性指数（SLEDAI）评分标准分为活动组（56例）和缓解组（48例）,比较各组结果的差异,并分析SLEDAI与IL-10和IL-18的相关性。结果 SLE组IL-10和IL-18水平分别为（18.25±3.66）、（582.61±65.28）pg/ml,明显高于对照组的（7.12±2.36）、（186.24±60.39）pg/ml,差异有统计学意义（P〈0.01）;SLE活动组IL-10和IL-18水平分别为（25.98±4.75）、（683.72±62.48）pg/ml,高于缓解组的（14.67±3.21）、（493.51±69.17）pg/ml,差异亦有统计学意义（P〈0.01）;SLE患者血清IL-10和IL-18水平与SLEDAI呈正相关（P〈0.05）。结论 IL-10和IL-18在SLE发病机制中发挥重要作用,而且与疾病活动性相关。     

IL-10RA基因多态性及其与系统性红斑狼疮关系的研究

目的:确定SLE模型小鼠IL-10RA基因变异及其与SLE表现型是否存在关联。方法:用微卫星遗传标记及数量性状位点(QTL)分析方法确定SLE模型小鼠B/W F1的SLE易感基因精确染色体定位并选取候选易感基因,对候选易感基因进行测序分析,选取有基因序列异常的候选易感基因进行PCR-SSCP分析,确定候选易感基因碱基序列变异位点与抗染色质抗体、抗DNA抗体,抗组蛋白抗体及蛋白尿等SLE表现型的相关关系。结果:QTL分析结果表明B/W F1×NZB小鼠抗染色质抗体易感基因与NZW型IL-10RA基因紧密连锁;测序分析发现IL-10RA基因编码区有18处碱基变异,其中7处碱基变异将导致编码氨基酸的变异;抗染色质抗体、抗DNA抗体,抗组蛋白抗体及蛋白尿等SLE表现型与NZW型IL-10RA基因密切相关。另一种SLE模型小鼠MRL的IL-10RA基因存在相同变异。结论:NZW小鼠IL-10RA基因编码区碱基序列存在变异,B/W F1×NZB小鼠SLE表现型与NZW小鼠第9染色体IL-10RA编码区碱基变异相关,提示IL-10RA可能是SLE模型小鼠的一个SLE易感基因。     

RANTES对肥大细胞系P815细胞分泌IL-10的诱导作用及其机制

目的检测RANTES对肥大细胞IL-10和IL-12分泌的影响和可能的信号转导通路。方法 P815肥大细胞培养、激发后收集不同时间点的细胞和上清液,用酶联免疫吸附试验(ELISA)检测上清中IL-10和IL-12的水平,用细胞激发信号ELISA(CASE)方法检测丝/苏氨酸蛋白激酶(Akt),细胞外信号调节激酶(ERK),信号转导子和转录激活子(STAT)3和p38丝裂原活化蛋白激酶信号转导通路蛋白磷酸化情况。结果 RANTES能够促进肥大细胞P815分泌IL-10而对IL-12的分泌无明显影响。LY294002能够阻断RANTES引起的肥大细胞IL-10分泌并抑制RANTES引起的Akt磷酸化。结论 RANTES刺激肥大细胞P815分泌IL-10可能是通过激活Akt信号转导通路实现的。     

Linkage of a major quantitative trait locus to Yaa gene-induced lupus-like nephritis in (NZW × C57BL/6)F1 mice

In the present study, we mapped the major quantitative trait loci (QTL) differing between the NZW and C57BL / 6 inbred strains of mice by making use of (NZW × C57BL / 6.Yaa)F1 mice, a model in which the lupus-like autoimmune syndrome observed in male mice is associated with the presence of an as yet unidentified Y chromosome-linked autoimmune acceleration gene, Yaa. Linkage analysis of 126 C57BL / 6 × (NZW × C57BL / 6.Yaa)F1 backcross males provided evidence for a major QTL on chromosome 7 controlling both the severity of glomerulonephritis and the production of IgG anti-DNA autoantibody and retroviral gp70-anti-gp70 immune complexes. Two additional QTL of C57BL / 6 origin on chromosome 17 had no apparent individual effects, but showed strong epistatic interaction with chromosome 7 QTL for disease severity and anti-DNA autoantibody production. Our data also identified on chromosome 13 a QTL of NZW origin with a major effect on the level of gp70, and showing an additive effect with the chromosome 7 QTL on the level of gp70 immune complexes. Our study thus provides a model to dissect the complex genetic interactions that result in manifestations of murine lupus-like disease.     

两条IL-6信号转导途径在人骨髓瘤细胞系U266中的相互拮抗作用

目的 研究人骨髓瘤细胞系U266中白细胞介素6（IL－6）信号转导途径及彼此之间的相互调控方式。方法 首先采用凝胶阻滞电泳（electrophoretic mobility shift assay,EMSA)方法观察分别参与两条IL－6信号转导途径的转录因子STAT3和NF－IL－6在U266细胞中的诱导激活状态并确定该细胞中的IL－6信号转导通路;继而采用转基因或化学试剂处理方法,特异性上调或下调其中一条IL－6信号途径的化,并同时观察另外一信号途径的激活状态。结果 1、以上两种转录因子分别参与的IL－6信号转导途径－－JAK／STAT和Ras/NF-IL-6,它们都能够在U266细胞中诱导激活;2、在高剂量IL－6（1－100ng/ml）刺激范围之内,一条IL－6信号途径活化水平的升高可同时导致另一条信号途径活化水平的下降。结论 在一定IL－6刺激剂量范围内,U266细胞中两条IL－6信号途径的诱导激活存在着相互拮抗作用。     

Evaluation of the effect of interleukin-10 on the multiplication of Mycobacterium avium complex in human macrophages and in C57BL/6 mice

Objectives: To examine the effect of recombinant human IL-10 (rhIL-10) on MAC infection of human macrophages and C57BL/6 mice.
Methods: We compared rhIL-10 with the effects of the immunosuppressive drugs prednisolone and cyciosporin A, both in vitro and in vivo.
Results: There was no effect of rhIL-10 on the multiplication of MAC in human macrophages after 1 week of infection. In C57BL/6 mice, rhIL-10 at 2.5 or 25 μg/mouse had no additional multiplicatory effect after 3 weeks of infection, while the spleens of mice treated with prednisolone had 600% higher bacteria than controls or rhIL-10-treated mice ( p <0.01).
Conclusions: These data suggest that rhIL-10 does not further decrease the resistance of human macrophages and C57BL/6 mice to MAC infection, whereas prednisolone leads to increased multiplication of MAC in the spleens of infected C57BL/6 mice. These results may be of interest in the context of the therapeutic use of rhIL-10 in some autoimmune disorders.     

The effect of interleukin 6 deficiency on myocardial signal transduction pathways activation induced by bacterial lipopolysaccharide in young and old mice

PurposeExaggerated release of proinflammatory mediators during sepsis contributes to inadequate vasodilatation and depressed myocardial contractility, which lead to development of shock and circulatory collapse. The aim of the study was to evaluate the effect of IL-6 and aging on activation of intracellular signaling pathways in the myocardium induced by bacterial lipopolysaccharide (LPS) administration.Material/methodsLPS was injected intraperitoneally to male 3- and 24-month old mice with systemic IL-6 gene knock-out (IL-6KO) and the reference strain (WT). LPS was given intraperitoneally in single low (0.1 mg/kg) or high (10 mg/kg) dose, or in two doses (0.1 + 10 mg/kg) with 24-h delay. The expression and phosphorylation of STAT3, ERK1/2, Akt1/2/3 proteins in the left ventricular myocardium was evaluated after 24 h using Western blotting.ResultsLow LPS dose induced higher STAT3 phosphorylation only in old IL-6KO mice, not affecting ERK1/2 and Akt1/2/3 phosphorylation in any group. High LPS dose upregulated STAT3 phosphorylation similarly in all groups, reduced ERK1/2 expression in young WT mice and upregulated Akt1/2/3 expression and phosphorylation in young IL-6KO mice. Pretreatment with low LPS dose attenuated phosphorylation of STAT3 in both old groups and phosphorylation of Akt1/2/3 in young IL-6KO group. Two-dose approach also significantly potentiated ERK1/2 phosphorylation in both old groups.ConclusionsObtained results show that IL-6 deficiency alters the activity of intracellular signaling pathways: JAK/STAT in old and Akt in young LPS-treated mice. This may indicate that lack of IL-6 attenuates Akt-related cytoprotective effect of pretreatment with low LPS dose in young but not in aged animals.     

Selective silencing of DNA-specific B lymphocytes delays lupus activity in MRL/lpr mice

The pathological DNA-specific B lymphocytes in lupus are logical targets for a selected therapeutic intervention. We have hypothesized that it should be possible to suppress selectively the activity of these B cells in lupus mice by administering to them an artificial molecule that cross-links their surface immunoglobulins with the inhibitory FcgammaIIb surface receptors. A hybrid molecule was constructed by coupling the DNA-mimicking DWEYSVWLSN peptide to a monoclonal anti-mouse FcgammaRIIb antibody. This chimeric antibody was added to cultured spleen cells from sick MRL/lpr mice, immunized with diphtheria toxoid, resulting in reduction of the numbers of anti-DNA but not of anti-diphtheria IgG antibody-producing cells. Intravenous infusions with the DNA-peptide antibody chimera to 7-wk-old animals prevented the appearance of IgG anti-DNA antibodies and of albuminuria in the next 2 months. The administration of the DNA-peptide chimeric antibody to 18 wk-old mice with full-blown disease resulted in the maintenance of a flat level of IgG anti-DNA antibodies and in delay of the aggravation of the lupus glomerulonephritis. The use of chimeric antibodies targeting inhibitory B lymphocyte receptors represents a novel approach for the selective suppression of autoreactive disease-associated B cells in autoimmune diseases.     

Phenotype conversion from rheumatoid arthritis to systemic lupus erythematosus by introduction of Yaa mutation into FcγRIIB‐deficient C57BL/6 mice

We previously established an IgG Fc receptor IIB (FcγRIIB)‐deficient C57BL/6 (B6)‐congenic mouse strain (KO1), which spontaneously develops rheumatoid arthritis (RA), but not systemic lupus erythematosus (SLE). Here, we show that when Y chromosome‐linked autoimmune acceleration (Yaa) mutation was introduced in KO1 strain (KO1.Yaa), the majority of KO1.Yaa mice did not develop RA, but instead did develop SLE. This phenotype conversion did not depend on autoantibody specificity, since KO1.Yaa mice, compared with KO1, showed a marked increase in serum levels of both lupus‐related and RA‐related autoantibodies. The increase in frequencies of CD69⁺ activated B cells and T cells, and the spontaneous splenic GC formation with T follicular helper cell generation were manifest early in life of KO1.Yaa, but not KO1 and B6.Yaa, mice. Activated CD4⁺ T cells from KO1.Yaa mice showed upregulated production of IL‐21 and IL‐10, compared with the finding in KO1 mice, indicating the possibility that this aberrant cytokine milieu relates to the disease phenotype conversion. Thus, our model is useful to clarify the shared and the disease‐specific mechanisms underlying the clinically distinct systemic autoimmune diseases RA and SLE.     

雷公藤碱戊对系统性红斑狼疮患者B细胞免疫功能的体外实验研究

目的：探讨雷公藤碱戊的免疫药理作用。方法：采用系统性红斑狼疮（ＳＬＥ）病人外周血单个核细胞（ＰＢＭＣ）体外培养观察雷公藤碱戊对ＳＬＥ病人ＰＢＭＣ的Ｂ细胞功能的影响及时效关系。结果：雷公藤碱戊能抑制ＳＬＥ的ＰＢＭＣ自发性增殖反应以及正常人和ＳＬＥ病人ＰＢＭＣ对ＳＡＣ刺激后的增殖反应。同时在培养的７２ｈ前加入雷公藤碱戊均能对正常人ＰＢＭＣ在ＰＷＭ诱导下的ＩｇＧ产生呈现抑制效应。结论：雷公藤碱戊对Ｂ细胞功能的多个环节具有抑制作用，其抑制Ｂ细胞产生ＩｇＧ的作用环节可能在于抑制了Ｂ细胞的活化增殖阶段