FasL与TNF引起凋亡的研究进展

死亡分子与死亡分子受体相互作用引发的细胞凋亡在保持自身平稳发挥着无法代替的重要作用。目前ＦａｓＬ,ＴＮＦ和ＴＲＡ１三个死亡分子及ＦａｓＴＮＦＲ,ＤＲ３／ＷＳ１－１和ＣＡＲ１四个死亡分子受体已被确定,本文就ＦａｓＬ和ＴＮＦ两个死亡分子在与其受体Ｆａｓ下调免疫反应,效应分子和免疫特赦的研究现状予以综述。     

白血病中Fas／FasL系统诱导的细胞凋亡作用

Fas／FasL系统是研究最深入的介导细胞凋亡的途径，当Fas与FasL结合时，可直接引起Fas^ 细胞的凋亡。但是在白血病的发病过程中，尽管高表达Fas，似乎仍存在着血细胞的凋亡不足。许多研究表明，白血病细胞对Fas／FasL途径介导的凋亡不敏感或存在抵抗也是导致白血病耐药的原因之一，白血病的Fas抗性与多种因素有关，对它们的研究可为白血病的治疗提供新的方法。     

FasL与TNF引起凋亡的研究进展

死亡分子与死亡分子受体相互作用引发的细胞凋亡在保持自身平稳发挥着无法代替的重要作用。目前ＦａｓＬ、ＴＮＦ和ＴＲＡ１ 三个死亡分子及Ｆａｓ、ＴＮＦＲ、ＤＲ３／ＷＳ１１ 和ＣＡＲ１ 四个死亡分子受体已被确定。本文就ＦａｓＬ和ＴＮＦ两个死亡分子在与其受体、Ｆａｓ 下调免疫反应、效应分子和免疫特赦的研究现状予以综述。     

环磷酰胺诱导的Fas／FasL依赖的鼠胸腺细胞凋亡

环磷酰胺(Cyclophosphamide,CPA)是具有抗肿瘤及免疫抑制双重药理作用的药物。研究已发现其对肿瘤细胞的杀伤作用是通过诱导细胞调亡而实现的[1],胸腺作为中枢性免疫器官,在免疫疫系统中起重要作用,本研究探讨CPA能否在体内诱导大鼠胸腺细胞调亡及其作用途径。1　材料和方法1.1　材料　雄性SD大鼠,重375～400g,白求恩医科大学动物中心喂养。细胞调亡原位末端标记试剂盒为美国Oncor公司产品,P53、Fas多克隆抗体及ABC试剂盒为Vector公司产品。环磷酰胺为奥地利ASTA药厂产品。1.2　动物处理　实验分两大组:第一组大鼠经灌胃给予0、2、7…     

白血病中Fas/FasL系统诱导的细胞凋亡作用

Fas FasL系统是研究最深入的介导细胞凋亡的途径 ,当Fas与FasL结合时 ,可直接引起Fas+ 细胞的凋亡。但是在白血病的发病过程中 ,尽管高表达Fas,似乎仍存在着血细胞的凋亡不足。许多研究表明 ,白血病细胞对Fas FasL途径介导的凋亡不敏感或存在抵抗也是导致白血病耐药的原因之一 ,白血病的Fas抗性与多种因素有关 ,对它们的研究可为白血病的治疗提供新的方法     

FasL诱导EAE小鼠淋巴细胞凋亡的实验研究

为了解FasL在诱导EAE淋巴细胞凋亡中发挥的作用 ,我们用髓鞘碱性蛋白 (MBP )致敏小鼠 ,建立动物模型 实验性自身免疫性脑脊髓膜炎 (EAE )。分析了EAE动物淋巴细胞膜上Fas及FasL分子的表达水平 ,检测了分泌的细胞因子 ;用FasL分子诱导了EAE淋巴细胞凋亡。结果表明 ,应用MBP致敏KM小鼠 ,成功地建立了EAE模型 ,KM小鼠的发病率为6 8 3% ,发病程度为 1～ 2分 ;小鼠特异性淋转比对照组高 ,并与发病评分成正比 ;细胞因子分泌水平提示以Th2为主 ;EAE淋巴细胞Fas和FasL的表达及凋亡显著高于对照组 ;FasL诱导淋巴细胞凋亡剂量依赖曲线表明 ,在一个较小的浓度范围内呈现正相关 ,若加入单抗可部分阻断FasL诱导的凋亡。结果提示Fas FasL在EAE动物淋巴细胞凋亡中起了重要作用。     

FasL基因转染对大鼠肥大细胞凋亡的影响

目的：观察FasL基因转染对大鼠肥大细胞凋亡的影响。方法：RT-PCR法扩增大鼠FasL穿膜区和胞外区cDNA，成功构建pcDNA3．1／FasL真核表达质粒，瞬时转染RBL-2H3，RT-PCR、免疫印迹法鉴定FasL在RBL-2H3上的表达，Annexinv流式细胞检测pcDNA3．1／FasL转染RBL-2H3后细胞的凋亡情况。结果：获得FasL穿膜区和胞外区eDNA，构建pcDNA3．1／FasL真核表达质粒，瞬时转染RBL-2H3后在细胞膜和上清均有FasL的存在，瞬时转染RBL-2H3后48小时，细胞发生凋亡。结论：吧大细胞可以通过Fas-FasL途径凋亡，为FasL基因应用于过敏性疾病的治疗提供依据。     

多发性硬化中Fas—FasL介导细胞凋亡研究进展

Fas和FasL结合后,通过信号转导可以诱导细胞凋亡。在多发性硬化中,Fas-FasL介导了自身反应性T细胞的凋亡,也介导了其它非T细胞的凋亡,如少突胶质细胞、星形细胞等。不少学者从多发性硬化的动物模型－实验性变态反应性脑脊髓膜炎（EAE）到病人进行了大量的研究。本文综述近年来这方面的最新研究进展。     

Fas／FasL与凋亡研究进展

Fas/FasL是凋亡(Apoptosis)诱导家族的重要成员之一,是一组调节组织发育和体内平衡的重要分子,其自身的分泌受到许多病理和生理因素的影响。已知有多种Fas变型基因的存在,表达可溶性Fas(sFas),与自身免疫性疾病和肿瘤等多种疾病有密切关系。     

心肌缺血损伤大鼠心肌细胞凋亡与Fas、FasL基因表达的变化

目的探讨异丙肾上腺素(isoprenaline,ISO)诱导的心肌缺血损伤大鼠心肌细胞凋亡与Fas和FasL基因表达变化。方法健康成年SD大鼠随机分为对照组及心肌缺血损伤组(损伤组),腹腔大剂量注射异丙肾上腺素,造成心肌缺血损伤模型。采用原位末端缺口标记(TUNEL)法观察心肌细胞凋亡的改变;应用Western blotting和免疫组化方法检测Fas和FasL蛋白表达的变化,并使用HPIAS10009图像分析仪进行定量分析。结果与对照组比较,缺血损伤组大鼠心肌细胞凋亡率明显增加(P<0.01)、心肌组织Fas、FasL蛋白表达水平显著升高(P<0.01)。结论心肌细胞凋亡及Fas、FasL基因过表达可能参与了心肌缺血损伤的发生和发展。     

Purification of poliovirus by affinity chromatography.

Poliovirus type 1 (Mahoney strain) has been purified by retention on a Sepharose 4B-antibody column and elution with 3M K thiocyanate. The virus was recovered in excellent yield and its purity was as high as that achieved by detergent treatment followed by sucrose gradient centrifugation. The column could be re-used and its capacity was sufficiently high to make it a useful method for the purification of milligram quantities of the virus.     

Molecular and Cellular Mechanisms RegulatingTand B Cell Apoptosis through Fas/FasL Interaction

Fas (CD95) and Fas ligand (FasL) are a receptor/ligand pair critically involved in lymphocyte homeostasis and peripheral tolerance such that genetic defect in either Fas or FasL results in an autoimmune lymphoproliferative syndrome. Fas is a type I transmembrane protein and a member of the tumor necrosis factor receptor (TNFR) family whereas FasL is a type II transmembrane protein and a member of TNF family. Binding of Fas by FasL induces apoptosis of the Fas-expressing cells. In the past few years. Fas/FasL interaction has been connected to a series of important phenomena previously viewed as independent immune processes. The activation-induced T cell death (AICD) and the FasL-mediated cytotoxicity by activated T cells are two critical mechanisms that can account for most of these phenomena. It is in the context of the two mechanisms that we discuss in this review the molecular and cellular events that occur during T/T and T/B interactions that account for the down-regulation of the immune response. We have also discussed recent advances in the areas of FasL gene regulation, lymphokine regulation of AICD, and regulation of B cell susceptibility to FasL. Investigation in these areas should help elucidate the role of Fas/FasL in the complex network of regulatory mechanisms that control immune response and autoimmunity.     

介绍一种大量纯化钙调素的方法

本文介绍了一种适于大量制备钙调素的简易实用方法。，我们用该方法从２．５ｋｇ猪脑组织中一次纯化了１５０ｍｇ，经鉴定，所提ＣａＭ的纯度在ＳＤＳ ＰＡＧＥ中呈一区带，分子量为１９ｋＤ，等电点为４．３８。并对ＰＤＥ有明显的激活作用，该作用可被特异性抑制剂ＴＦＰ抑制。     

Correlation between expression of DcR3 on tumor cells and sensitivity to FasL

To investigate the correlation between sensitivity to Fas ligand （FasL） and expression level of decoy receptor 3 （DcR3） on tumor cell surface, Fas/DcR3 mRNA expression was detected by RT-PCR. Anti-DcR3 mAb was used to detect expression level of DcR3 on surface of tumor cells by flow cytometry. Caspase-8, caspase-9, caspase-3, Bcl-2 expressions were analyzed by Western blot, respectively. Sensitivity to apoptosis induced by FasL was determined by Annexin V apoptosis kit. The expressions of DcR3 on the surface of tumor cells from high to low were approximately 35.3% in BGC823 cells, and 21.6% in MCF-7 cells, respectively. The apoptotic rates induced by FasL from low to high were 15.6% in BGC823 cells, and 58.2% in MCF-7 cells, respectively. There was a significant correlation between the expression levels of DcR3 with FasL-inducing apoptosis. Cellular ＆ Molecular Immunology.     

APO—1／Fas及其配体介导超抗原SEB诱导的淋巴细胞凋亡

为了探讨超抗原诱导的淋巴细胞凋亡机制．本实验采用超抗原金黄色葡萄球菌肠毒素B（SEB）处理人外周血单个核细胞,采用流式细胞术检测细胞DNA断裂的百分率;同时动态观察了细胞表面APO-1、FasL的表达及细胞内Bcl－2蛋白的表达。结果表明,SEB不但可引起人外周血淋巴细胞凋亡,而且可诱导淋巴细胞表达APO-1和FasL。细胞凋亡的百分率与细胞表面APO-1和FasL的表达呈正相关（r＝0．71,r＝0．98）,与细胞内Bcl－2表达呈负相关（r＝－0．72）。由此可见,APO－1／Fas及其配体参与了超抗原SEB诱导的淋巴细胞凋亡。     

FasL/Fas pathway is involved in dengue virus induced apoptosis of the vascular endothelial cells

The hallmark of the dengue hemorrhagic fever/dengue shock syndrome is hematologic abnormality. The pathogenesis of dengue hemorrhagic fever/dengue shock syndrome remains unknown. Our work showed that the dengue virus serotype‐2 induced apoptosis in human umbilical vein endothelial cells. Fas (CD95), Tumor Necrosis Factor receptors, and Tumor Necrosis Factor‐related apoptosis‐inducing ligand receptors are the most common death receptors, which can induce apoptosis. Compared with the untreated human umbilical vein endothelial cells, Fas expression was increased both in the mRNA level and on the surface of infected human umbilical vein endothelial cells. FasL was expressed at similar levels on human umbilical vein endothelial cells over a course of dengue virus serotype‐2 infection, but the expression in mRNA level was increased in infected human umbilical vein endothelial cells. It is possible that there is soluble FasL secreted from human umbilical vein endothelial cells in the supernatant. Tumor Necrosis Factor‐related apoptosis‐inducing ligand receptor 1 and Tumor Necrosis Factor receptors 1–2 were constantly very low, whereas Tumor Necrosis Factor‐related apoptosis‐inducing ligand receptors 2–4 decreased after dengue virus serotype‐2 infection. This result suggested that dengue virus serotype‐2 may inhibit Tumor Necrosis Factor‐related apoptosis‐inducing ligand receptors‐induced apoptosis. The apoptotic rates in human umbilical vein endothelial cells were decreased upon the addition of caspase family inhibitors. In addition, activated caspase 8 and caspase 3 were also observed by Western blot following dengue virus serotype‐2 infection. Thus, it is shown that the Fas/FasL pathway may participate in dengue virus‐induced apoptosis of vascular endothelial cells in vitro. J. Med. Virol. 82:1392–1399, 2010. © 2010 Wiley‐Liss, Inc.     

用膜亲和层析法纯化小鼠腹水中的单克隆抗体

目的 用膜亲和层析法（MAC）纯化小鼠腹水中单克隆抗体（mAb)。方法 采用尼龙滤膜ZBM作为介质吸附抗人血清白蛋白（HSA）,将含mAb的腹水滤过此ZBM,再将吸附于ZBM上的mAb解离下来,获得纯化的mAb。结果 直径50mm吸附了HSA的ZＢＭ。经１０mL腹水循环滤过两次,即可达到最大mAb结合量。从ＺＢＭ上解离mAb,仅需解离液滤过１次就可完成,纯化的mAb经ＰＡＧＥ检测,只显示１条带。用纯化的mAb做金标记斑点免疫渗滤法（ＤＩＧＦＡ）检测ＨＳＡ标准品时,其灵敏度较用未纯化的腹水时提高了２０倍,结论 用尼龙滤膜ＺＢＭ改进的膜亲和层析法,可用于纯化中的单克隆抗体 且简便、省时、有效。     

反义RNA阻断Hep G2细胞FasL的表达及功能研究

应用分子克隆技术将人FasLcDNA片段反向插入逆转录病毒载体pLXSN ,构建重组质粒pL (hFasL AS )SN ,转染包装细胞PA317后获得FasL反义RNA重组逆转录病毒表达载体假病毒上清 ,经NIH3T3细胞检测其感染滴度后转染肝癌细胞株HepG2细胞并筛选建系 ,命名为HepG2 hFasL AS。半定量RT PCR检测显示HepG2 FasL AS细胞FasL的mRNA明显少于正常HepG2细胞 ;FACS检测显示HepG2 FasL AS细胞FasL的表达与HepG2细胞相比显著下降 ;同时 ,HepG2 FasL AS细胞导致HL 6 0细胞凋亡能力有所下降。表明人FasL反义RNA重组逆转录病毒表达载体能抑制转染细胞FasL的表达并下调其致凋亡功能