TRAIL及其受体在正常心脏组织的免疫组织化学定位

目的 :观察正常人心脏组织中 TRAIL 及其受体 DR4、DR5、Dc R1和 Dc R2的表达分布情况 ,为进一步明确这些分子在心肌损伤中的作用机制提供形态学依据。方法 :用抗 TRAIL及其受体的单克隆抗体 （m Ab） ,应用免疫组织化学染色技术 ,检测正常人心脏组织中 TRAIL及其受体 DR4、DR5、Dc R1和 Dc R2的表达。结果 :TRAIL及其受体 DR4、DR5、Dc R1和 Dc R2在正常人心脏组织中均有表达 ,表达产物主要分布于胞浆内 ,胞膜上也有表达。结论 :TRAIL及其受体可能参与某些病理状态下心肌细胞的凋亡过程     

主要组织相容性复合物在特发性炎性肌病中的表达及临床应用价值

目的 研究多发性肌炎(PM)和皮肌炎(DM)患者骨骼肌组织中的主要组织相容性复合物(MHC)的表达,探讨它们在特发性炎性肌病(IIM)发病机制中的作用及临床应用价值.方法 选取我院1986-2007年风湿免疫科收住的IIM患者骨骼肌组织标本45例(PM 19例,DM 26例),以性别、年龄相匹配的外伤患者骨骼肌标本30例作为对照.用免疫组织化学方法检测人类白细胞抗原(HIA)-A/B/C(即MHC-Ⅰ分子)和HLA-DR(即MHC-Ⅱ分子)在PM/DM患者及对照组肌组织中的表达.结果 HLA-A/B/C在18例PM、24例DM和3例对照组肌组织中有表达,阳性率分别为95%、92%和10%;HLA-DR在PM组、DM组和对照组肌组织中的阳性表达率分别为84%、81%和13%.HLA-A/B/C和HLA-DR在PM/DM患者肌组织中的表达量均较正常对照组明显升高(P均<0.05),但在PM组和DM组间差异无统计学意义(P均>0.05).根据组织病理学特点把PM/DM患者分为病变程度不同的3组,HLA-A/B/C和HLA-DR在PM/DM患者肌组织中的表达量与肌纤维变性、坏死、炎症浸润的程度和各项临床及实验室指标无明显关联(P均>0.05).结论 MHC-Ⅰ和Ⅱ类分子在PM/DM患者肌肉组织中表达增高,在无明显炎症浸润和肌纤维坏死的患者中也可见到这种表达的上调,检测肌组织中MHC-Ⅰ和Ⅱ类分子的表达可用于PM/DM的诊断.     

死亡受体5和死亡诱骗受体2在腰椎间盘组织中的表达及意义

目的 探讨死亡受体5(DR5)、死亡诱骗受体2(DCR2)在腰椎间盘中的表达及意义.方法 选取腰椎间盘突出症行椎间盘组织摘除术的患者73例作为观察组;另外寻找非正常死亡成人15例取腰椎间盘32个.采用PV6000两步免疫组织化学染色法对两组腰椎间盘进行检测.结果 观察组髓核及纤维环内的DR5细胞阳性率明显高于对照组,两组比较差异明显(P＜0.05)；DCR2细胞阳性率两组比较差异不明显,无统计学意义(P＞0.05).结论 DR5可能参与了腰椎间盘组织的细胞凋亡过程.     

肿瘤坏死因子相关的凋亡诱导配体受体在白血病细胞中的表达及其意义

目的研究肿瘤坏死因子相关的凋亡诱导配体(TRAIL)受体在白血病细胞中的表达及其临床意义。方法应用RT-PCR方法对76例白血病患者的骨髓,包括15例急性淋巴细胞性白血病(急淋)、10例急性淋巴细胞性白血病缓解期(急淋缓解)、16例急性非淋巴细胞性白血病(急非淋)、11例急性非淋巴细胞性白血病缓解期(急非淋缓解)、12例慢性粒细胞性白血病急性变期(慢粒急变期)和12例慢性粒细胞性白血病慢性期(慢粒慢性期)以及25例正常人骨髓或外周血白细胞表面TRAIL受体的表达进行检测。结果急淋、急淋缓解、急非淋、急非淋缓解、慢粒急变期和慢粒慢性期患者骨髓的白细胞表面死亡受体(deadreceptor,DR)DR4和DR5表达高,而诱骗受体(decoyreceptor,DcR)DcR1和DcR2表达低,且DR4的表达高于DR5的表达;缓解期DR4和DR5的表达高于患者组;正常人骨髓或外周血白细胞中DR4和DR5表达低,而DcR1和DcR2表达高。结论TRAIL受体在不同类型的白血病细胞中的表达具有明显的差异性。     

肺癌组织中肿瘤坏死因子相关凋亡诱导配体受体的表达及其临床意义

目的　探讨不同类型肿瘤坏死因子相关凋亡诱导配体 (TRAILR )在非小细胞肺癌(NSCLC)中的表达及其临床意义。方法　采用RT PCR检测 40例肺癌组织、对应癌旁非癌肺组织以及非小细胞肺癌细胞株A5 49不同类型TRAILR的表达 ,并结合临床资料进行分析。结果　 40例肺癌组织表达死亡受体DR4,3 5例肺癌组织表达死亡受体DR5 ;40例癌旁肺组织均表达死亡受体DR5和DR4,3 3例肺癌组织不表达诱捕受体DcR1,2 5例肺癌组织不表达DcR2 ,而 40例癌旁肺组织均表达DcR。肺癌细胞株A5 49中有DR5、DR4、DcR2的表达 ,但DcR1表达缺失。肺癌组织中DR的表达水平与肿瘤的分期有关 ,Ⅲ期肿瘤DR的表达显著低于Ⅰ /Ⅱ期DR表达 (P <0 .0 5 )。DR的表达与病人的性别、年龄、病理学分型无关。结论　肺癌存在TRAILR类型的表达差异 ,诱捕受体 (特别是DcR1)的表达缺失为TRAIL治疗肺癌提供了靶点     

MHC-Ⅱ联合MHC-Ⅰ免疫组化染色在特发性炎性肌病诊断中的应用价值探讨

目的 探讨主要组织相容性复合体(MHC)-Ⅱ联合MHC-Ⅰ免疫组化染色在特发性炎性肌病(IIM)诊断中的应用价值。方法 收集2010年3月至2018年4月在首都医科大学宣武医院神经内科行肌肉活检患者的标本29份,并通过医院电子病历系统收集患者的临床资料,包括4种IIM[皮肌炎(DM)5例,多发性肌炎(PM)5例,散发性包涵体肌炎(IBM)4例及坏死性自身免疫性肌病(NAM)5例]和2种非炎性肌病(NIM)[肌营养不良(MD)5例,dysferlinopathy肌病5例]。将标本进行苏木精-伊红(HE)染色及MHC-Ⅰ、MHC-Ⅱ免疫组化染色。结果 免疫组化染色结果显示,PM、DM及IBM患者肌肉标本MHC-Ⅰ阳性率达100.0%,NAM和MD患者肌肉标本MHC-Ⅰ阳性率为80.0%,dysferlinopathy肌病患者肌肉标本MHC-Ⅰ阳性率为20.0%。PM和IBM患者肌肉标本MHC-Ⅱ阳性率分别为40.0%、50.0%,其余类型疾病患者肌肉标本的MHC-Ⅱ免疫组化染色均呈阴性。结论 相较于MHC-Ⅰ免疫组化染色,MHC-Ⅱ免疫组化染色在鉴别IIM与NIM中有较高的特异性。MHC-...     

特发性炎性肌病骨骼肌炎症浸润的组织病理学分析

目的：研究多发性肌炎（PM）和皮肌炎（DM）患者骨骼肌组织中浸润的炎性细胞的类型、分布和定位，探讨它们在特发性炎性肌病（IIM）发病机制中的作用。方法；选取IIM患者肌肉组织标本45例，其中PM19例，DM26例。用免疫组织化学方法检测PM／DM患者骨骼肌组织中CD3、CD4、CD8、CD20、CD45RA和CD45RO分子的表达。结果：PM组肌组织的损伤主要表现为肌纤维坏死和肌内膜炎，而DM组主要表现为束周萎缩和血管周围炎。CD3、CD8和CD45RO分子在PM和DM患者肌组织中均有阳性表达，两组间表达量差异无统计学意义（P均〉0．05），主要分布在PM／DM患者肌组织的肌内膜和血管周围处。而CD20、CD4、CD45RA分子在DM组的阳性表达量较PM组明显增多（P均〈O．05），且主要分布在血管周围。结论：细胞免疫和体液免疫均参与了PM和DM的发病，而体液免疫在DM的发病中可能起更主要的作用。     

5-FU增强TRAIL诱导的胃癌BGC823细胞凋亡的实验研究

目的 观察5-FU对TRAIL诱导的胃癌BGC823细胞凋亡的影响,明确死亡受体5(DR5)在5-FU和TRAIL诱导凋亡中的作用.方法 采用MTT法测定细胞活力、流式细胞仪检测细胞凋亡、免疫印迹检测蛋白表达.结果 TRAIL可导致BGC823细胞轻度的增殖抑制和少量的细胞凋亡.与单药TRAIL和5-FU相比,TRAIL联合5-FU对细胞的增殖抑制和诱导凋亡作用明显增强(P＜0.05).免疫印迹结果显示,TRAIL没有改变DR5的蛋白表达,而5-FU作用BGC823细胞48 h后,DR5蛋白表达上调(P＜0.05).TRAIL和5-FU联合作用后,DR5蛋白表达同样明显上调(P均＜0.05).结论 5-FU通过上调DR5蛋白表达提高了BGC823细胞对TRAIL的敏感性.     

西黄丸含药血清增加TRAIL对RBE细胞的抑瘤作用及其机制

[目的]探讨西黄丸含药血清与肿瘤坏死因子相关细胞凋亡诱导配体(TNF related apoptosis inducing ligand,TRAIL)单独或联合对人胆管癌细胞RBE增殖和凋亡的影响及其机制。[方法]制备高、中、低剂量西黄丸含药血清;人胆管癌细胞RBE分别用西黄丸含药血清、人重组可溶性TRAIL蛋白(rsTRAIL)及两者联合处理;MTT法检测细胞增殖;流式细胞术检测细胞凋亡;分别用免疫荧光染色结合流式细胞术和RT-PCR检测TRAIL R1/DR4和TRAIL R2/DR5在细胞表面及mRNA水平的表达。[结果]高剂量西黄丸含药血清可以抑制RBE细胞增殖,诱导其凋亡,较对照组差异有统计学意义(P0.01);中、低剂量西黄丸含药血清未见上述抑瘤作用;rsTRAIL以剂量依赖性方式抑制RBE细胞增殖,诱导其凋亡;低剂量西黄丸含药血清联合rsTRAIL处理后,RBE细胞增殖抑制率和凋亡率明显高于单用rsTRAIL组,比较差异有统计学意义(P0.05)。低剂量西黄丸含药血清单独或联合rsTRAIL作用于RBE细胞36h后,死亡受体TRAIL R1/DR4和TRAIL R2/DR5在细胞表面和mRNA水平的表达均较对照组细胞明显增强,差异均有统计学意义(P0.01)。[结论]西黄丸含药血清增强TRAIL对人胆管癌细胞RBE的增殖抑制和凋亡诱导效应,其机制可能与西黄丸含药血清上调RBE细胞TRAIL死亡受体表达相关。     

Differential expression of IFN-alpha and TRAIL/DR5 in lymphoid tissue of progressor versus nonprogressor HIV-1-infected patients

Loss of CD4+ T cells, the hallmark of HIV pathogenesis, was suggested to be partly due to apoptosis. We recently reported that IFN-alpha produced by HIV-1-activated plasmacytoid dendritic cells (pDCs) contributes to CD4+ T cell apoptosis by the TNF-related apoptosis-inducing ligand (TRAIL)/death receptor (DR)5 pathway. Here, we show that HIV-1-induced intracellular expression of IFN-alpha in pDCs is coupled to increased expression of IFN regulatory factor 7 and MyD88 by pDCs in vivo and in vitro. Expression of IFN-alpha was increased in lymphoid tonsillar tissue (LT) of patients with progressive (HIV(prog)) compared with nonprogressive (HIV(NP)) HIV-1 disease and to uninfected controls. LT from HIV(prog) exhibited higher TRAIL and DR5 mRNA levels than LT from HIV(NP) or controls. TRAIL mRNA levels in LT correlated with plasma viral load. We show that HIV-1 induces IFN-alpha and the TRAIL/DR5 apoptotic pathway in LT, suggesting a role for these cytokines in HIV-1 immunopathogenesis.     

Expression of TNF-related apoptosis-inducing Ligand receptors and antitumor tumor effects of TNF-related apoptosis-inducing Ligand in human hepatocellular carcinoma

AIM: To investigate the expression of TNF-related apoptosis -inducing Ligand (TRAIL) receptors and antitumor effects of TRAIL in hepatocellular carcinoma (HCC). METHODS: Expression of TRAIL receptors was determined in 60 HCC tissues, 20 normal liver samples and two HCC cell lines (HepG2 and SMMC-7721). The effects of TRAIL on promoting apoptosis in HCC cell lines were analyzed after the cells were exposed to the recombinant TRAIL protein, as well as transfected with TRAIL-expression construct. In vivo effects of TRAIL on tumor growth were investigated by using nude mice HCC model of hepG2. RESULTS: Both death receptors were expressed in all HCC tissues and normal hepatic samples. In contrast, 54 HCC tissues did not express DcR1 and 25 did not express DcR2. But both DcR were detectable in all of the normal liver tissues. The expression patterns of DR and DcR in HCC samples (higher DR expression level and lower DcR expression level) were quite different from those in normal tissue. DR5, DR4, and DcR2 expressed in both cell lines, while no DcR1 expression was detected. Recombinant TRAIL alone was found to have a slight activity as it killed a maximum of 15 % of HCC cells within 24 h. Transfection of the TRAIL cDNA failed to induce extensive apoptosis in HCC lines. In vivo administration of TRAIL gene could not inhibit tumor growth in nude mice HCC model. However, chemotherapeutic agents or anticancer cytokines dramatically augmented TRAIL-induced apoptosis in HCC cell lines. CONCLUSION: Loss of DcR (especially DcR1) in HCC may contribute to antitumor effects of TRAIL to HCC.HCC is insensitive towards TRAIL-mediated apoptosis, suggesting that the presence of mediators can inhibit the TRAIL cell-death-inducing pathway in HCC. TRAIL and chemotherapeutic agents or anticancer cytokines combination may be a novel strategy for the treatment of HCC.     

Aortic valvular interstitial cells apoptosis and calcification are mediated by TNF-related apoptosis-inducing ligand

Background/Objectives

Calcific aortic valvular disease (CAVD) is an actively regulated process characterized by the activation of specific osteogenic signaling pathways and apoptosis. We evaluated the involvement in CAVD of the TNF-related apoptosis-inducing ligand (TRAIL), an apoptotic molecule which induces apoptosis by interacting with the death receptor (DR)-4 and DR5, and whose activity is modulated by the decoy receptor (DcR)-1 and DcR2.

Methods

Sections of calcific and normal aortic valves, obtained at surgery time, were subjected to immunohistochemistry and confocal microscopy for TRAIL immunostaining. Valvular interstitial cells (VICs) isolated from calcific (C-VICs) and normal (N-VICs) aortic valves were investigated for the gene and protein expression of TRAIL receptors. Cell viability was assayed by MTT. Von Kossa staining was performed to verify C-VIC ability to produce mineralized nodules. TRAIL serum levels were detected by ELISA.

Results

Higher levels of TRAIL were detected in calcific aortic valves and in sera from the same patients respect to controls. C-VICs express significantly higher mRNA and protein levels of DR4, DR5, DcR1, DcR2 and Runx2 compared to N-VICs. C-VICs and N-VICs, cultured in osteogenic medium, express significantly higher mRNA levels of DR4, Runx2 and Osteocalcin compared to baseline. C-VICs and N-VICs were sensitive to TRAIL-apoptotic effect at baseline and after osteogenic differentiation, as demonstrated by MTT assay and caspase-3 activation. TRAIL enhanced mineralized matrix nodule synthesis by C-VICs cultured in osteogenic medium.

Conclusions

TRAIL is characteristically present within calcific aortic valves, and mediates the calcification of aortic valve interstitial cells in culture through mechanism involving apoptosis.     

IL-1RA对急性排斥期大鼠角膜移植物TRAIL及其受体DR4表达的影响

目的研究白细胞介素-1受体拮抗剂(IL-1RA)对急性排斥期角膜移植物肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其死亡受体-4(DR4)表达的影响。方法建立大鼠同种异体穿透角膜移植模型,设生理盐水处理组和IL-1RA治疗组。免疫组织化学法检测急性排斥期角膜植片TRAIL及DR1表达,并以正常大鼠角膜作对照。结果TRAIL及DR4在正常角膜均有表达,主要分布于上皮层。急性排斥期角膜植片各层TRAIL及DR4表达均增高;与IL-1RA治疗组相比．生理盐水处理组TRAIL及DR4表达增高更明显。结论TRAIL及DR4的表达参与角膜移植免疫排斥反应的发生,IL-1RA可通过调节其表达抑制角膜移植免疫排斥反应。     

Identification of tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) and its receptors in adult rat ventral prostate

Tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor-alpha (TNF-alpha) family of cytokines that is known to induce apoptosis upon binding to its death domain-containing receptors, DR4/TRAIL-R1 and DR5/TRAIL-R2. Two additional TRAIL receptors, DcR1/TRAIL-R3 and DcR2/TRAIL-R4, lack functional death domains and act as decoy receptors for TRAIL. In this study, the presence of TRAIL and its receptors was investigated in adult rat hormonosensitive ventral prostate. TRAIL and its receptors were identified in the rat ventral prostate in terms of protein and mRNA. TRAIL and its receptors were immunolocalized in prostatic epithelial cells.     

CD4+ T-cell death induced by infectious and noninfectious HIV-1: role of type 1 interferon-dependent, TRAIL/DR5-mediated apoptosis

It has been proposed that direct and indirect mechanisms contribute to the unresolved issue of CD4(+) T-cell depletion that results from HIV-1 infection. We recently reported that plasma levels of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) are elevated in HIV-1-infected patients and that they correlate with viral load. The present study investigates the expression of TRAIL death receptor 5 (DR5) in the peripheral-blood mononuclear cells (PBMCs) of HIV-1-infected patients and its role in CD4(+) T-cell death. DR5 expression was elevated and associated with the apoptotic marker annexin V. Apoptosis was reduced in CD4(+) T cells when cultured with anti-DR5 antibody. CD4(+), but not CD8(+), T cells from uninfected donors expressed TRAIL, DR5, and activated caspase-3 when cultured with infectious or noninfectious HIV-1, resulting in preferential apoptosis of CD4(+) T cells. TRAIL, caspase-3 expression, and apoptosis were type 1 interferon (IFN) dependent. Induction of apoptosis and DR5 expression required glycoprotein 120 (gp120)-CD4 interaction. Finally, we analyzed DR5 expression by CD4(+) T cells in highly active antiretroviral therapy (HAART)-treated patients. The decreased viral loads and increased CD4 counts of HAART-responsive patients were associated with a decrease in DR5 mRNA expression by CD4(+) T lymphocytes. We propose a novel model in which a type 1 IFN-regulated TRAIL /DR5 mechanism induces apoptosis of HIV-1-exposed CD4(+) T cells.