应用椭偏光学显微成像对IL-6与其受体的相互应用的初步观察

白细胞介素6(IL-6)是一种具有复杂生物功能的细胞因子,可由多种淋巴类和非淋巴类细胞产生.它对机体多种组织及细胞均有不同程度的作用[1-3].近年来发现,临床上免疫异常性疾病,如发热、淋巴结肿大、血沉增快、急性期蛋白增高、高γ球蛋白血症、自身抗体阳性等症状都与IL-6的异常表达密切相关.IL-6的生物活性是通过细胞膜表面特异性受体介导的[4].研究IL6与其受体的相互作用对于揭示某些疾病的发病机制,监测疾病进程以及指导临床治疗等均具有重要意义.     

信号转导和转录激活因子STAT6的结构与功能

信息转导和转录激活因子 6 (ｓｉｇｎａｌｔｒａｎｓ ｄｕｃｅｒａｎｄａｃｔｉｖａｔｏｒｏｆｔｒａｎｓｃｒｉｐｔｉｏｎ 6 ,ＳＴＡＴ6 )是信号转导和转录激活因子 (ＳＴＡＴ)家族的一员 ,是一种 94ｋＤ的蛋白质。它通过ＪＡＫ ＳＴＡＴ信号转导途径介导细胞因子ＩＬ 3、ＩＬ 4及ＩＬ 1 3诱导的基因表达 ,特别是在促进ＩＬ 4诱导的ＭＨＣⅡ类抗原及免疫球蛋白受体等的表达[1] 及Ｔｈ2细胞和淋巴细胞的发育[2 ] 上发挥了重要的作用。ＳＴＡＴ6的作用机制大致包括了ＳＴＡＴ6的激活、ＳＴＡＴ6的二聚化与核转运及与ＤＮＡ的结合诱导基因的…     

白介素-11对肠粘膜损伤的防治作用

白介素11(ＩＬ11)是一种多功能的细胞因子,ＩＬ11及其受体的广泛组织分布特点,提示它可能具有多种重要生理功能。最初有关ＩＬ11生理作用的研究主要集中在其对造血功能、Ｂ细胞免疫功能及神经系统发育的调节作用等方面。近年的一些研究资料表明,ＩＬ11对肠粘膜损伤具有明显的防治作用,初步显示ＩＬ11在肠粘膜损伤防治的临床方面具有潜在的应用价值。肠粘膜由于其快速增殖、不断自我更新的特点,化疗药物、射线和烧伤等因素极易引起肠粘膜损伤,破坏肠粘膜的正常屏障功能,导致肠源性感染机会增加。Ｄｕ等[1]发现,ＩＬ11可显著提高接受放化疗联…     

TRPC6在肺部的生理与病理生理功能

经典瞬时受体电位(transient receptor potential canonical,TRPC)通道是一类非选择性钙离子通道,TRPC6是TRPC家族的成员之一,其基因编码的蛋白在人体脑、肾、肝和肺等多个器官均有表达。TRPC6不仅是肺中表达最丰富的TRPC通道,也是目前肺部疾病中研究最多的TRPC通道。TRPC6参与动物体内许多重要的生理和病理调节过程,TRPC6的基因突变或过表达可引起胞内钙离子信号通路异常,从而导致多种病理生理改变。肺部疾病的病理生理过程有多种细胞共同参与。TRPC6在不同的细胞中表达不同,表现的生理与病理生理功能则有差异。     

CD147在调节性T细胞中作用的研究进展

CD147是一种免疫球蛋白超家族,它与肿瘤侵袭和转染,类风湿性关节炎,病毒入侵,淋巴细胞的迁移和活化,老年痴呆症,疟原虫入侵等病理过程密切先关,已成为癌症等疾病的新型药物靶标分子。作为一个单次跨膜蛋白,CD147的多功能性依赖于它的胞外结构域以及细胞膜和细胞外多种分子的相互作用。有研究发现CD147可作为细胞膜受体并且在上皮细胞,肿瘤细胞和免疫T细胞中均有表达。其可作为T细胞亲环素受体并具有趋化T细胞的功能。近几年研究发现,CD147在调节性T细胞高表达并且具有标志性意义。该文就以作为活性调节性T细胞标志的CD147的潜在作用进展综述如下。     

生长因子及其受体在淋巴瘤中异常表达的研究进展

生长因子是一类与受体结合后可以促进细胞增殖和调节细胞多项功能的多肽分子。生长因子及其受体信号通路包括Ras/MAPK、PI3K/AKT和STAT等不仅调控正常细胞的生物学行为,对恶性肿瘤细胞增殖、分化、转化和迁移也具有重要意义。研究发现多种生长因子如VEGF、PDGF和IGF及其受体在多种实体肿瘤如肺癌、乳腺癌、结肠癌中发现有异常表达,在淋巴瘤如DLBCL、PTCL、ML和NL中也存在异常的共同表达,提示在淋巴瘤中可能构成生长因子及其受体的自分泌/旁分泌环路。生长因子及其受体的表达对淋巴瘤患者的预后有一定指导意义,临床研究发现表达生长因子或其受体阳性患者比表达阴性患者有较差的临床预后。这可能与生长因子及其受体对淋巴瘤细胞的增殖、转移和耐药调控有关。目前生长因子及其受体已成为潜在的药物靶点,多种生长因子及其受体抑制剂在开发和临床试验中。本文就近年来生长因子及其受体在淋巴瘤中异常表达研究进展作简要综述。     

蛇毒酶制剂在脑疾病中应用的概况

20世纪 70年代以来 ,我国的蛇毒制剂研究进展很快 ,于 80年代开始应用于临床 ,尤其是脑疾病的应用最多 ,均取得良好的治疗效果。蛇毒的成分很复杂 ,含有多种蛋白质、十几种酶、几种非酶活性蛋白质或多肽素及少量无机离子和其他有机物 ,因此 ,蛇毒具有多种多样的毒理和药理效应 [1 ,2 ]。经过分离纯化制成一种有效的抗凝溶栓制剂 ,已用于临床的有 :去纤酶、蝮蛇抗栓酶、抗栓酶 - 3、精制蝮蛇抗栓酶、蕲蛇酶和降纤酶 ,其主要成份是类同的 ,均具有以下药理作用 [3～ 7] :(1 )类凝血酶 :蛇毒含有类凝血酶样酶 ,能直接作用于纤维蛋白原 ,使其降解…     

人白细胞介素22基因的分离及其在大肠杆菌中的克隆与表达

白细胞介素 2 2 (ＩＬ 2 2 )是 2 0 0 0年发现的一种新的人类细胞因子 ,它在多种组织中均有表达 ,包括胰腺、脑 ,肝、小肠、直肠和肾等 .在功能方面 ,ＩＬ 2 2上调肝细胞急性期产物 ,参与炎症反应[1～ 3] ;诱导胰腺相关蛋白ＰＡＰ1 Ｒｅｇ2和ＯＰＮ的高表达 ,表明ＩＬ 2 2参与胰腺免疫功能反应[4 ] ;它对ＣｏｎＡ刺激下Ｔｈ1细胞ＩＦＮ γ的产生没有明显的抑制作用 ,却大大抑制了Ｔｈ2细胞ＩＬ 4的分泌 ,这对于哮喘有潜在的治疗作用[1] .我们利用反转录ＰＣＲ获得了编码ｈＩＬ 2 2成熟蛋白的ｃＤＮＡ ,并构建了高表达载体ｐＢＶｈＩＬ 2 2 ,…     

拟南芥类受体胞质激酶RBK2的生物信息学分析

[目的]对拟南芥类受体胞质激酶RBK2(ROP binding protein kinases 2)的结构、分子进化和基因表达等进行预测分析。[方法]利用生物信息学在线相关数据库和分析软件对目的蛋白的基本理化性质、二级及三级结构、同源性、基因表达、蛋白定位、相互作用蛋白进行分析预测。[结果]拟南芥类受体胞质激酶RBK2是一个亲水性不稳定蛋白,主要构成原件为α螺旋及β片状折叠,进化树构建及多序列比对显示其与琴叶拟南芥中名为激酶家族蛋白的蛋白进化距离最近。主要存在于细胞质和细胞核中,其编码基因在成熟花粉中强烈表达,且RBK2与PP2C家族蛋白有较强的相互作用。[结论]拟南芥RBK2很可能通过参与信号转导途径在植物种子萌发、花粉成熟及细胞分裂等方面发挥重要作用。     

RANTES生物功能的多样性

调节活化正常Ｔ细胞表达与分泌的趋化因子(ｒｅｇｕｌａｔｅｄｕｐｏｎａｃｔｉｖａｔｉｏｎｎｏｒｍａｌＴｃｅｌｌｅｘｐｒｅｓｓｅｄａｎｄｓｅｃｒｅｔｅｄ ,ＲＡＮＴＥＳ)为ＣＣ类趋化蛋白 ,具有多种不同的生物学功能。它不仅对多种细胞有趋化作用 ,还可以强烈的激活淋巴细胞 ,参与炎症反应的发生 ,调节细胞的生长和分化 ,参与ＨＩＶ的感染。1.ＲＡＮＴＥＳ及其受体ＲＡＮＴＥＳ仅含 6 8个氨基酸残基 ,其来源广泛 ,ＮＫ细胞[1] 以及γδ Ｔ细胞[2 ] 均可组成性地表达ＲＡＮＴＥＳ ;炎症反应中ＣＤ8 Ｔ细胞、上皮细胞、成纤维细胞和血小…     

IL-6 Trans-Signaling via the Soluble IL-6 Receptor: Importance for the Pro-Inflammatory Activities of IL-6

Interleukin-6 (IL-6) is a cytokine with many activities. It has functions in the regulation of the immune system and the nervous system. Furthermore, IL-6 is involved in liver regeneration and in the metabolic control of the body. On target cells, IL-6 binds to an 80 kDa IL-6 receptor (IL-6R). The complex of IL-6 and IL-6R associates with a second protein, gp130, which thereupon dimerizes and initiates intracellular signaling. Whereas gp130 is expressed on all cells, IL-6R is only present on few cells in the body including hepatocytes and some leukocytes. Cells, which do not express IL-6R cannot respond to the cytokine, since gp130 alone has no measurable affinity for IL-6. Interestingly, a soluble form of IL-6R (sIL-6R) comprising the extracellular portion of the receptor can bind IL-6 with a similar affinity as the membrane bound IL-6R. The complex of IL-6 and sIL-6R can bind to gp130 on cells, which do not express the IL-6R, and which are unresponsive to IL-6. This process has been called trans-signaling. Here I will review published evidence that IL-6 trans-signaling is pro-inflammatory whereas classic IL-6 signaling via the membrane bound IL-6R is needed for regenerative or anti-inflammatory activities of the cytokine. Furthermore, the detailed knowledge of IL-6 biology has important consequences for therapeutic strategies aimed at the blockade of the cytokine IL-6.     

人IL-6 /sIL-6R 结合分子模型的建立及在药物筛选中的应用

目的：建立人IL-6 /sIL-6R 结合的分子模型,用于筛选IL-6 /sIL-6R的抑制剂。方法：将人IL-6基因克隆至原核表达载体pET28a（+）中表达IL-6蛋白,western blot及人IL-6检测试剂盒分析鉴定表达蛋白。同法将人sIL-6R在pET15b载体中表达,纯化并用western blot检测目的蛋白。依据ELISA原理建立IL-6 /sIL-6R 结合的分子模型,并通过改变IL-6、sIL-6R及IL-6 antibody的浓度来优化该模型,用于IL-6 /sIL-6R拮抗药物的筛选。结果：人IL-6可在载体PET28a（+）中高效表达,且经western blot鉴定正确,人IL-6检测试剂盒检测显示具有较高的免疫活性。sIL-6R在PET15b中表达,western blot鉴定正确。通过对IL-6 /sIL-6R结合的分子模型的优化,得到其最佳条件为：IL-6R 1?g/well, IL-6 500ng/well, IL-6 antibody 1?g/well。应用该模型筛选发现有些化合物可显著抑制IL-6与其受体的结合。结论：成功构建IL-6 /sIL-6R结合的分子模型,为高通量筛选IL-6拮抗剂提供平台。     

The involvement of IL-6 and IL-8 in acute invasive gastroenteritis of children

The involvement of the proinflammatory cytokines, interleukin 8 (IL-8) and 6 (IL-6), was studied during the first 72 h of acute invasive gastroenteritis. Study population included 33 infants and young children aged six months to six years and seven age-matched controls. As a group, patients with acute invasive gastroenteritis had an increased serum level of IL-8 and IL-6 as compared with healthy controls (p < 0.002 and p < 0.001, respectively). Subjects were then divided into two groups based on stool cultures (proven and non-proven bacterial cultures). Patients with bacterial-proven acute invasive gastroenteritis tended to have increased IL-8 serum concentrations (p < 0.07) as compared with those with non-proven bacterial etiologies and IL-6 levels were only detected in subjects with positive bacterial cultures (p < 0.05). When dividing each sub-group into early and late blood drawing with respect to disease onset, no statistical differences were found in each group but subjects with bacterial-proven etiologies had significant higher IL-6 levels as compared with non-proven etiologies at the two time points (p < 0.019 and p < 0.015, respectively).In conclusion, the proinflammatory cytokines, IL-6 and IL-8, are involved in acute invasive gastroenteritis. The difference in IL-6, and to a lesser degree IL-8, between proven and non-proven bacterial etiologies, needs further investigation.     

肺炎嗜衣原体、IL-6与冠心病关系的血清学探讨

采用血清学的方法观察冠心病(CHD)与肺炎嗜衣原体感染及白细胞介素-6之间相关性并对其致病机理作一简要探讨。用酶联免疫吸附分析(ELISA)技术检测136例CHD患者血清中的肺炎嗜衣原体抗体CP-IgA、CP-IgG、CP-IgM阳性率,并对其中肺炎嗜衣原体阳性者进行IL-6的检测。有64%CHD患者血清特异性CP-IgA呈阳性,与健康对照组的8.8%阳性率差异显著(P<0.01);而CP-IgG和CP-IgM与对照组的阳性率无显著差异(P>0.05);CHD肺炎嗜衣原体CP-IgA阳性者的IL-6明显高于对照组,两者之间有显著的差异(P<0.01)。冠心病病人血清肺炎嗜衣原体抗IgA的高阳性率与冠心病之间存在着有意义的联系,是CP-IgA慢性感染CP的标记物,肺炎嗜衣原体感染参与了冠心病的发生与发展。而炎症因子IL-6水平的升高也提示炎症反应在冠心病的发生以及疾病的发展过程中起着重要的作用。     

Novel Insights into Interleukin 6 (IL-6) Cis- and Trans-signaling Pathways by Differentially Manipulating the Assembly of the IL-6 Signaling Complex

The IL-6 signaling complex is described as a hexamer, formed by the association of two IL-6·IL-6 receptor (IL-6R)·gp130 trimers, with gp130 being the signal transducer inducing cis- and trans-mediated signaling via a membrane-bound or soluble form of the IL-6R, respectively. 25F10 is an anti-mouse IL-6R mAb that binds to both membrane-bound IL-6R and soluble IL-6R with the unique property of specifically inhibiting trans-mediated signaling events. In this study, epitope mapping revealed that 25F10 interacts at site IIb of IL-6R but allows the binding of IL-6 to the IL-6R and the recruitment of gp130, forming a trimer complex. Binding of 25F10 to IL-6R prevented the formation of the hexameric complex obligate for trans-mediated signaling, suggesting that the cis- and trans-modes of IL-6 signaling adopt different mechanisms for receptor complex assembly. To study this phenomenon also in the human system, we developed NI-1201, a mAb that targets, in the human IL-6R sequence, the epitope recognized by 25F10 for mice. Interestingly, NI-1201, however, did not selectively inhibit human IL-6 trans-signaling, although both mAbs produced beneficial outcomes in conditions of exacerbated IL-6 as compared with a site I-directed mAb. These findings shed light on the complexity of IL-6 signaling. First, triggering cis- versus trans-mediated IL-6 signaling occurs via distinctive mechanisms for receptor complex assembly in mice. Second, the formation of the receptor complex leading to cis- and trans-signaling biology in mice and humans is different, and this should be taken into account when developing strategies to inhibit IL-6 clinically.     

Anti-interleukin 6 (IL-6) receptor antibody suppresses Castleman's disease like symptoms emerged in IL-6 transgenic mice

Transgenic mice carrying human IL-6 cDNA fused with a murine major histocompatibility class-I promoter (H-2L(d)) were serially administered with anti-interleukin-6 receptor (IL-6R) monoclonal antibody (mAb), MR16-1, from the age of 4 weeks to estimate its efficacy on a variety of disorders developed in these mice, most of which are similar to the disorders associated with Castleman's disease. In the control mice treated with isotype-matched mAb, a massive and multiple IgG1 plasmacytosis, mesangial proliferative glomerulonephritis, leukocytosis, thrombocytosis, anemia and abnormalities of blood chemical parameters have developed in accordance with the elevation of serum IL-6, and 50% of mice have died of renal failure by 18 weeks of age. In contrast, the treatment with MR16-1 prevented all these symptoms and prolonged the lifetime of the majority of the mice. Thus, the constitutive overexpression of IL-6 caused various disorders, and the treatment with anti-IL-6R mAb completely prevented from these symptoms. These results clearly confirm that IL-6 indeed plays an essential role in the pathogenesis of a variety of disorders. Furthermore, anti-IL-6R mAb could provide novel therapy for Castleman's disease and MR16-1 should be a useful tool to estimate therapeutic potential of IL-6 antagonists in a variety of murine models for human disease.     

Theoretical investigation of IL-6 multiprotein receptor assembly

Interleukin-6 (IL-6) is a multifunctional cytokine that regulates cell growth, differentiation, and cellular functions in many cell lineages. Recently, evidences for the formation of an active hexameric complex with an IL-6:IL-6Rα:gp130 stoichiometry of 2:2:2 have been obtained by different experimental approaches. Analysis of the electrostatic potential complementarity between IL-6 and its receptors has been used, in this study, to guide the assembly of homology-based 3D models of the components. The results strongly support a mechanism whereby the active cytokine (IL-6:IL-6Rα) associates with the signal transducing gp130 protein, and the trimeric complex formed further dimerizes to form the hexameric species. Furthermore, computational simulations of the multiprotein complexes provide a rationalization of data from mutation experiments and highlight some key protein–protein interactions which have not yet been the subject of mutagenesis studies. Proteins 29:528–544, 1997. © 1997 Wiley-Liss, Inc.     

慢病毒载体介导shRNA干扰IL-6Rα基因表达削弱IL-6对猪脂肪细胞分化的影响

为研究白细胞介素-6(IL-6)对猪脂肪细胞分化的影响及其分子机制,构建猪IL 6Rα基因RNA干扰（RNA interference, RNAi）慢病毒载体;用IL-6Rα-RNAi重组慢病毒预处理原代培养的猪前体脂肪细胞或不处理, 然后用100 ng/mL IL-6处理分化第6 d的脂肪细胞48 h．通过测定甘油释放量检测脂肪细胞的脂解率;油红O染色提取法测定脂肪细胞的脂质含量;采用RT-PCR 和Western印迹检测脂肪细胞分化相关基因的mRNA 和蛋白表达．结果显示,IL-6显著抑制猪脂肪细胞分化,并下调PPARγ2、Perilipin A和IRS-1的mRNA及蛋白表达,同时增强ERK1/2磷酸化;IL-6Rα-RNAi预处理前体脂肪细胞则显著逆转IL 6的上述作用．总之,IL-6通过多重机制抑制猪脂肪细胞分化;而且本研究构建的IL-6Rα-RNAi重组慢病毒载体可有效阻断IL-6信号,为进一步研究IL-6的功能奠定了基础．     

IL-6在代谢类疾病及癌症中的功能研究进展

由于白细胞介素IL-6在炎症中广泛存在,所以其通常被视为一种促炎细胞因子,与肿瘤坏死因子TNF及IL-1β在炎症中具有同等作用。IL-6能够通过刺激活化B细胞的增殖而分泌抗体,刺激T细胞的增殖及细胞毒性T淋巴细胞（CTL）的活化,刺激肝细胞合成急性期蛋白而参与炎症反应,并能促进血细胞发育。此外,有研究指出IL-6 在多种生理条件下包括分辨炎症功能中具有更大的作用。综述了IL-6信号调控炎症相关的癌症及代谢性疾病如肥胖症、肝脏代谢、骨骼肌代谢及运动中复杂生物学功能的机理,从多方面阐述了该多效性因子的意义,以期为IL-6在疾病治疗中的应用提供参考。     

TGF-β is necessary for induction of IL-23R and Th17 differentiation by IL-6 and IL-23

TGF-β and IL-6 induce Th17 differentiation, and IL-23 is required for expansion and maintenance of Th17 cells. Recently, it was shown that IL-6 up-regulates IL-23R mRNA in naive CD4⁺ T cells and therefore IL-6 and IL-23 synergistically promote Th17 differentiation. However, the molecular mechanism whereby IL-6 and IL-23 induce Th17 differentiation and the relevance to TGF-β remain unknown. Here, we found that IL-6 up-regulated IL-23R mRNA expression, and IL-6 and IL-23 synergistically augmented its protein expression. The combination induced Th17 differentiation, and TGF-β1 further enhanced it. IL-6 augmented endogenous TGF-β1 mRNA expression, whereas the amount of TGF-β produced was not enough to induce Th17 differentiation by IL-6 alone. However, unexpectedly, the up-regulation of IL-23R and induction of Th17 differentiation by IL-6 and IL-23 were almost completely inhibited by anti-TGF-β. These results suggest that the induction of IL-23R and Th17 differentiation by IL-6 and IL-23 is mediated through endogenously produced TGF-β.