基质细胞衍生因子-1和血小板第4因子对体外扩增后脐血CD34+细胞黏附特性和趋化功能的影响

为了研究基质细胞衍生因子-1（SDF—1）和血小板第4因子（PF4）对扩增后脐血CD34^＋细胞归巢相关功能的影响，将纯化的脐血CD34^＋细胞接种入无血清培养液中，加入不同组合的细胞因子FST（FL＋SCF＋TPO）、FST＋SDF—1、FST＋PF4或FST＋SDF—1＋PF4，分别于培养第7、10、14天检测CD34^＋细胞扩增倍数、集落形成能力、细胞的黏附分子表达、总黏附性、趋化功能。结果表明：①加入SDF—1的实验组CD34^＋细胞及造血祖细胞集落扩增倍数高于对照组；②加入SDF—1明显上调扩增的CD34^＋细胞CD49e的表达，加入PF4明显上调扩增的CD34^＋细胞CD49e、CD54的表达，在扩增体系中加入SDF—1或PF4均能够明显提高扩增的CD34^＋细胞的总黏附性；③在扩增体系中加入SDF—1能够明显提高扩增的CD34^＋细胞的自发迁移率，但导致CXCR-4的表达和SDF—1诱导迁移率降低；而PF4能够明显提高扩增的CD34^＋细胞的CXCR-4的表达和SDF—1诱导迁移率；在扩增体系中同时加入SDF—1和PF4能够明显提高扩增的CD34^＋细胞自发迁移率和SDF—1诱导迁移率。结论：体外扩增体系中加入SDF—1和PF4能够上调部分归巢相关黏附分子的表达，保持扩增的CD34^＋细胞的黏附和迁移能力，有利于降低体外扩增对造血干／祖细胞（HSPC）归巢相关功能的不利影响，维持扩增的HSPC的归巢潜能。     

VCAM-1及ICAM-1在胃癌组织中的表达及临床意义

目的 观察胃癌组织中血管细胞黏附分子（VCAM-1）及细胞间黏附分子（ICAM-1）的表达，探讨其临床意义。方法 用免疫组化法（S—P法）测定67例胃癌患者胃癌组织和非癌组织VCAM-1及ICAM-1的表达，对胃癌标本进行系统病理学检查。结果 胃癌患者胃癌组织VCAM-1及ICAM-1表达显著高于非癌组织。胃癌组织VCAM-1及ICAM—1表达水平明显与临床病理、淋巴结转移及TNM分期有关。结论 胃癌组织VCAM-1及ICAM-1的表达可能反映了肿瘤的发生和转移，具有一定的临床意义。     

全反式维甲酸诱导急性早幼粒细胞白血病细胞分化为树突状细胞的实验研究

全反式维甲酸（ATRA）诱导APL细胞分化为成熟树突状细胞（dendritic cell，DC），目前国内外鲜见报道。本研究探讨全反式维甲酸与经典细胞因子共同作用诱导急性早幼粒细胞白血病（APL）细胞分化为DC的可能性，为研制APL-DC疫苗提供新的方法。对初治APL患者骨髓单个核细胞及HL-60细胞株分别在完全培养液中培养，用经典细胞因子组合（GM—CSF、IL-4、TNF-α）共处理，ATRA只在实验组加用。观测细胞形态，检测DC免疫表型，测定上清液IL-12水平，观察MLR反应及CTL效应。结果表明：实验组所得细胞有较明显树突状突起，有APL遗传学特征，其CD1a、CD83、CD80、CD86、HLA—DR和CD1d表达及IL-12分泌水平明显增高，并具有明显的MLR反应及CTL效应，但在HL60-DC中，CD1a增加无统计学差异。结论：利用ATRA可以成功诱导APL细胞为成熟功能性DC，其介导的MLR及CTL效应明显。经ATRA组合诱导的树突状细胞CD1d表达明显增高，推测可能参与NKT细胞激活，具体机制需进一步研究。     

骨髓基质细胞表面CD40分子配基化促进IL-6和Flt3配体的释放及其作用

目的：分析人骨髓基质细胞（BMSC）CD40分子及其配体（CD40L）的表达，观察BMSC经CD40激发型单抗（5C11）激发后，其培养上清中IL－3、IL－6、Flt3配体（FL）和干细胞因子（SCF）含量变化及对纯化脐血CD34^ 细胞的作用。方法：新鲜分离人骨髓单个核细胞，经体外培养获得BMSC。间接免疫荧光标记，流式细胞仪分析细胞表面CD40和CD40L的表达，并与5C11（20μg/ml）共同孵育，分别于24，48和72h取上清液，ELISA法测定IL－3、IL－6、FL和SCF含量；在与5C11共同培养24h后，收取培养上清，与纯化的脐血CD34^ 细胞共同孵育，1周后进行细胞计数、细胞集落培养，并用钙磷脂结合蛋白（Annexin V)标记，流式细胞仪分析细胞凋亡。结果：发现BMSC表达CD40分子，经5C11激发后，BMSC IL－6和FL分泌增加，而对IL－3及SCF含量无影响。BMSC经5C11激发培养后支持脐血CD34^ 细胞的增殖，培养7d后其细胞数和集落数显著多于对照组，且细胞凋亡率低，二者差异有显著性（P＜0．01）。结论：BMSC CD40分子配基化可促进IL－6和FL的分泌，并支持CD34^ 细胞的增殖，CD40／CD40L可能通过调节细胞因子的释放参与造血调控。     

脐血CD34+细胞体外扩增时HOXB4基因表达的变化

同源盒基因家族成员HOXIM反映原始造血干／祖细胞（PHSC／PHPC）的自我更新和增殖能力。本研究采用实时定量RT—PCR的方法在mRNA水平上检测HOXIM基因的表达，以观察体外扩增脐血CD34^＋细胞自我更新的水平。结果显示：随着体外培养时间的延长，虽然CD34^＋细胞数增加，但HOXB4表达下降；周后，HOXB4几乎检测不到，与成熟外周血淋巴细胞表达HOXIM的水平相同；CD34^＋细胞与骨髓间充质干细胞（BM—MSC）共培养可以减缓CD34^＋细胞HOXIM表达的下降。结论：脐血CD34^＋细胞体外扩增过程中自我更新能力逐渐下降。CD34^＋与BM—MSC共培养有助于减缓体外扩增的CD34^＋细胞自我更新能力的丧失。     

VCAM-1、ICAM-1与造血干细胞的动员、归巢及作用机制

血管细胞黏附分子-1（VCAM-1）和细胞问黏附分子-1（ICAM-1）是目前研究较多的两种黏附分子,其在正常骨髓造血与非造血细胞的表达是构成细胞与细胞、细胞与基质相互识别与黏附的主要基础,可参与造血干／祖细胞的动员、归巢、增殖和分化等。对VCAM-1和ICAM-1在造血干细胞动员和归巢中的作用进行深入探讨,将为由此而进行的基础和临床研究奠定基础。     

LFA-1和VLA-4参与人高增殖潜能内皮祖细胞与血管内皮的黏附和跨内皮迁移

为了了解淋巴细胞功能相关抗原1（lymphocyte function—associated antigen1，LFA-1）和极迟反应抗原4（very lateantigen 4，VLA-4）在高增殖潜能内皮祖细胞（high proliferative potential endothelial progenitor cells，HPP—EPCs）归巢过程中与血管内皮的黏附和跨内皮迁移中的作用，利用流式细胞术检测HPP—EPC中整合蛋白B1和B2的表达以及小鼠骨髓内皮细胞相应的受体的表达。利用体外黏附和迁移实验研究经过功能级别的中和抗体阻断VLA-4和LFA-1后HPP—EPC黏附和迁移细胞数目的变化。结果表明，HPP—EPC表达整合蛋白B1和B2，活化后小鼠骨髓内皮细胞表达细胞间黏附分子1（intercellular adhesion molecule1，ICAM-1）和血管细胞黏附分子1（vascular cell adhesion molecule1，VCAM-1）；加CDlla抗体组黏附细胞或CD49d抗体组黏附和迁移细胞均较同型对照抗体组少，而且加CDlla和CD49d两种抗体联用组黏附和迁移细胞明显减少，其细胞数较任何单一抗体组少。结论：LFA-1和VLA-4在HPP—EPC与血管内皮的黏附和跨内皮迁移中发挥了重要的作用。     

急性CO中毒大鼠脑内VCAM-1表达CD4+T淋巴细胞浸润规律的研究及高压氧干预后的变化

目的 观察急性CO中毒后大鼠脑内血管内皮细胞黏附分子-1（VCAM-1）表达、CD4^＋T淋巴细胞浸润情况，从系统细胞免疫反应角度探讨迟发性脑病（DNS）与免疫反应的关系。方法 30只SD雄性大鼠随机分为对照组，染毒后1、3、7d组和高压氧（HBO）治疗7d组，每组6只。采用HE和免疫组织化学染色方法，观察染毒后各时间点大鼠脑内病理形态学变化，及VCAM-1表达、CD4^＋T淋巴细胞浸润情况。结果 对照组无VCAM-1表达、无CD4^＋T淋巴细胞浸润；各染毒组大鼠脑内微血管内皮细胞和变性坏死的神经元均有VCAM-1不同程度的表达。染毒后1d组为表达高峰（P〈0．01），3d组VCAM-1的表达数量有所减少。此时，局部大脑皮层开始有CD4^＋T淋巴细胞浸润；至染毒后7d，在部分变性坏死的神经元上仍有VCAM-1的持续表达，而CD4^＋T淋巴细胞浸润达高峰（P〈0．01）。染毒后立即进行HBO治疗7d，其VCAM-1、CD4^＋T淋巴细胞均显著减少，与染毒后7d组相比差异有统计学意义（P〈0．05）。结论 系统细胞免疫反应可能参与了DNS的发生。     

microRNA在人脐血CD34^＋CD38^-、CD34^＋CD38^＋细胞中的差异性表达

为进一步探讨microRNA（miRNA）在造血调控中的作用奠定基础,比较了miRNA在人脐血CD34^＋CD38^-、CD34^＋CD38^＋细胞中的差异性表达。从人脐血中分离单个核细胞（MNC）,应用FACSVantage分选流式细胞仪分选CD34^＋CD38^-、CD34^＋CD38^＋细胞;抽提miRNA后与miRNA芯片杂交,应用生物信息学技术分析miRNA芯片的表达结果。结果发现,miRNA在人脐血CD34^＋CD38^＋细胞的表达水平比在CD34^＋CD38^-细胞的表达水平降低至1／2以下者共11个,表达水平升高至2倍以上者共73个,以上84个miRNA被称为“干细胞性”miRNA。经比较Georgantas等和芯片表达结果,发现有12个（14、29％,12／84）相同的miRNA。部分miRNA经历了类似于CD34在造血细胞表面的发展历程。应用生物信息学技术可寻找到新的与造血调控相关的miRNA簇及miRNA的下游靶基因。结论：“干细胞性”miRNA在正常造血中发挥着重要作用,即miRNA的系统表达模式一造血干／祖细胞基因表达谱一造血干／祖细胞的自我更新和系列定向分化。     

重组人粒细胞集落刺激因子动员对淋巴细胞功能相关抗原1／细胞间黏附分子1信号途径诱导CD4^＋T淋巴细胞活化及增殖的影响

目的：观察在异基因外周血造血干，祖细胞移植中重组人粒细胞集落刺激因子动员对淋巴功能相关抗原1/细胞间黏附分子1信号途径诱导的外周血CD4＋T淋巴细胞的影响，验证重组人粒细胞集落刺激因子动员对淋巴细胞功能相关抗原1/细胞间黏附分子1信号通路的抑制作用。 方法：选择2006—06/2007—06在解放军总医院血液科异基因外周血造血干细胞移植前进行重组人粒细胞集落刺激因子动员的10例健康供者，对治疗方案均知情同意，且得到医院伦理道德委员会批准。给予重组人粒细胞集落刺激因子10μg，（kg·d）进行动员，在动员前1天和动员后第5天取供者外周静脉血，用miniMACS磁珠分选系统分离纯化CD4＋T淋巴细胞，分别用CD3单克隆抗体OKT3＋细胞间黏附分子1、佛波酯＋离子霉素刺激活化CD4^＋T淋巴细胞，用双色荧光标记检测动员前后CD4^＋T淋巴细胞激活后活化标记CD69，CD25的表达；用四甲基偶氮唑盐法检测动员前后OKT3＋细胞间粘黏附分子1、佛波酯＋离子霉素刺激CD4^＋T淋巴细胞增殖能力变化。 结果：①纯化后CD4^＋T淋巴细胞纯度均在90％以上。②动员后OKT3＋细胞间黏附分子1组、佛波酯＋离子霉素组CD4^＋T淋巴细胞活化标记CD69和CD25的表达均明显低于动员前（P〈0．01）。②动员后OKT3＋细胞间黏附分子1组、佛波酯＋离子霉素组CD4^＋T淋巴细胞增殖率均明显低于动员前（P〈0．05）。 结论：重组人粒细胞集落刺激因子动员可能抑制了淋巴功能相关抗原1/细胞间黏附分子1协同刺激信号，从而使CD4^＋T淋巴细胞活化、增殖能力下降。     

维甲酸对小鼠骨髓基质细胞粘附分子表达和粘附功能的影响

探讨体外维甲酸 (RA)对小鼠骨髓基质细胞 (BMSC)粘附分子ICAM 1和VCAM 1的表达及人脐血单个核细胞 (UCBMNC)对BMSC粘附率的影响。用流式细胞术方法检测 0 .1 ,1 .0和 1 0 .0 μmol/LRA处理后BMSC粘附分子ICAM 1和VCAM 1的表达 ,以MTT方法检测 3种浓度RA处理后UCBMNC对BMSC粘附率的改变。结果显示 ,1 .0和 1 0 .0 μmol/LRA能增加BMSC上ICAM 1的表达及UCBMNC对BMSC的粘附率。RA对VCAM 1的表达无明显影响 ;RA上调ICAM 1的表达与增加UCBMNC对BMSC的粘附率呈正相关 (r =0 .7883 ,P <0 .0 5)。结论 :体外RA能上调小鼠BMSC上ICAM 1的表达和增加UCBMNC对BMSC的粘附率 ,两者呈正相关 ,为阐明BMSC粘附分子与造血干细胞归巢的相关关系及RA促进脐血造血干细胞归巢提供了体外实验依据     

骨髓移植小鼠骨髓基质细胞内皮抑制素(endostatin)及血管细胞黏附分子-1(VCAM-1)表达相关性研究

本研究探讨内皮抑制素（endostatin）及血管细胞黏附分子-1（VCAM-1）在骨髓移植（BMT）小鼠骨髓基质细胞中表达的相关性及川芎嗪对其表达的影响。取健康BALB／c小鼠，随机分为3组：正常组（不做处理），骨髓移植对照组（简称生理盐水组）和骨髓移植＋川芎嗪治疗组（简称川芎嗪组）。生理盐水组和川芎嗪组分别胃饲生理盐水0．2毫升／只和川芎嗪注射液每次2毫影只，2次／天。在BMT后第7、14、21、28天处死小鼠，用间接免疫荧光法和RT—PCR法检测骨髓基质细胞endostatin mRNA及其蛋白、VCAM-1 mRNA及其蛋白的表达情况，并探讨二者表达的相关性。结果表明：endostatin蛋白主要表达于骨髓基质细胞核内，VCAM-1蛋白主要表达于细胞浆内。BMT后第7、14、21天川芎嗪组和生理盐水组endostatin mRNA和蛋白的表达水平显著低于正常组（P〈0．01或P〈0．05），且川芎嗪组低于生理盐水组（P〈0．01或P〈0．05）；到第28天生理盐水组已基本恢复正常，而川芎嗪组仍未恢复正常水平。BMT后第7、14、21天川芎嗪组和生理盐水组VCAM-1 mRNA和蛋白的表达均明显低于正常组（P〈0．01或P〈0．05），且川芎嗪组明显高于生理盐水组（P〈0．01或P〈0．05），到第28天，川芎嗪组VCAM-1的表达已基本恢复正常，而生理盐水组仍未恢复正常，两组之间有显著性差异（P〈0．01）。相关分析显示，生理盐水组骨髓基质细胞endostatin和VCAM-1 mRNA及其蛋白表达呈负相关（P〈0.01或P〈0．05），而川芎嗪组endostatin和VCAM-1 mRNA及其蛋白表达呈正相关（P〈0．01或P〈O．05）。结论：内皮抑制素可通过影响骨髓基质细胞表面VCAM-1的表达水平，阻碍基质细胞与造血细胞之间和细胞外基质与造血细胞之间的联系而影响骨髓造血；川芎嗪能提高BMT小鼠骨髓基质细胞表面黏附分子的表达，加快BMT后HSPC的归巢及在骨髓中的增殖，并通过反馈调节BMT小鼠骨髓基质细胞新血管生成抑制因子内皮抑制素的表达，促进骨髓微环境的修复，加速移植后骨髓的造血重建。     

Role of vascular cell adhesion molecule 1/very late activation antigen 4 and intercellular adhesion molecule 1/lymphocyte function-associated antigen 1 interactions in antigen-induced eosinophil and T cell recruitment into the tissue

To determine the role of vascular cell adhesion molecule 1 (VCAM- 1)/very late activation antigen 4 (VLA-4) and intercellular adhesion molecule 1 (ICAM-1)/lymphocyte function-associated antigen 1 (LFA-1) interactions in causing antigen-induced eosinophil and T cell recruitment into the tissue, we studied the effect of the in vivo blocking of VCAM-1, ICAM-1, VLA-4, and LFA-1 by pretreatment with monoclonal antibodies (mAb) to these four adhesion molecules on the eosinophil and T cell infiltration of the trachea induced by antigen inhalation in mice. The in vivo blocking of VCAM-1 and VLA-4, but not of ICAM-1 and LFA-1, prevented antigen-induced eosinophil infiltration into the mouse trachea. On the contrary, the in vivo blocking of VCAM-1 and VLA-4, but not of ICAM-1 and LFA-1, increased blood eosinophil counts after antigen challenge, but did not affect blood eosinophil counts without antigen challenge in sensitized mice. Furthermore, the expression of VCAM-1 but not ICAM-1 was strongly induced on the endothelium of the trachea after antigen challenge. In addition, pretreatment with anti-IL-4 mAb decreased the antigen-induced VCAM-1 expression only by 27% and had no significant effect on antigen-induced eosinophil infiltration into the trachea. The in vivo blocking of VCAM- 1 and VLA-4 inhibited antigen-induced CD4+ and CD8+ T cell infiltration into the trachea more potently than that of ICAM-1 and LFA-1. In contrast, regardless of antigen challenge, the in vivo blocking of LFA- 1, but not of ICAM-1, increased blood lymphocyte counts more than that of VCAM-1 and VLA-4. These results indicate that VCAM-1/VLA-4 interaction plays a predominant role in controlling antigen-induced eosinophil and T cell recruitment into the tissue and that the induction of VCAM-1 expression on the endothelium at the site of allergic inflammation regulates this eosinophil and T cell recruitment.     

内皮缩血管肽反义寡核苷酸促进骨髓移植小鼠造血重建

目的观察内皮缩血管肽(Endostatin)反义寡核苷酸转染骨髓基质细胞后在骨髓移植(BMT)小鼠骨髓造血恢复过程中的作用。方法以脂质体作为转染载体,转染不同剂量的 FITC 标记的 Endostatin 反义寡核苷酸,荧光倒置显微镜观察转染效率,用流式细胞术检测最佳转染条件下转染率,用 RT-PCR 和 Western blot 方法检测最佳转染条件下 Endostatin 反义寡核苷酸对 BMT 后不同时间点小鼠骨髓基质细胞 Endostatin mRNA 及其蛋白质和血管细胞间黏附分子1(VCAM-1)mRNA 及其蛋白表达水平的影响。实验分为4组:①正常组:未经任何处理组;②BMT 组:空白对照组;③BMT+转染Endostatin 反义寡核苷酸组;④BMT+转染 Endostatin 错义寡核苷酸组。结果①在体外成功地将 En-dostatin 反义寡核苷酸导入骨髓基质细胞,转染率达86%;②以 Endostatin 反义寡核苷酸转染骨髓基质细胞后,BMT 后不同时间点骨髓基质细胞 Endostatin mRNA 及其蛋白表达被显著抑制[Endostatin 的灰度值分别为(0.09±0.03)～(1.44±1.19)和(0.02±0.02)～(0.14±0.05)](P<0.01或P<0.05),表明转染成功;③Endostatin 反义寡核苷酸转染有效促进了 BMT 后不同时间骨髓基质细胞 VCAM-1mRNA 及其蛋白表达[VCAM-1的灰度值分别为(1.60±0.92)～(8.05±0.87)和(0.07±0.02)～(0.67±0.09)](P<0.01或P<0.05);④Endostatin 错义寡核苷酸对 BMT 后不同时间骨髓基质细胞 Endostatin 和 VCAM-1的表达基本无影响(P>0.05)。结论 Endostatin 反义寡核苷酸可降低Endostatin 表达,增强 VCAM-1的表达,从而促进骨髓微血管生成,改善基质细胞与造血细胞之间和细胞外基质与造血细胞之间的联系而影响骨髓造血。     

Adhesion molecule behavior during rejection and infection episodes after heart transplantation.

In cardiac transplant recipients the release of soluble cellular adhesion molecules intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-Selectin into serum is pronounced during immune activation. It is uncertain whether there is a specific pattern of release during infection or cardiac allograft rejection. In a prospective study, 30 consecutive cardiac allograft recipients were followed for a median period of 11.4 months (range 1-34). Soluble ICAM-1 (sICAM-1), soluble VCAM-1 (sVCAM-1) and soluble E-Selectin (sE-Selectin) were measured in addition to acute phase proteins (C-reactive protein, alpha1-antitrypsin), complement factors (C3, C4) and beta2-microglobulin. The measured serum levels were correlated with the clinical status of the transplant recipient: 1) uneventful clinical status; 2) asymptomatic infection; 3) symptomatic infection and 4) rejection. Forty age-matched healthy subjects served as controls. Six days before biopsy-proven cardiac allograft rejection sICAM-1-release started to increase (p < 0.05) as compared to uneventful clinical status. The peak concentration of sICAM-1 was measured three days before rejection. On the day of rejection, serum concentrations of sICAM-1 (p < 0.001) and sVCAM-1 (p < 0.05) were increased, whereas sE-Selectin was not markedly elevated. In symptomatic infections, the serum concentrations of sICAM-1 (p < 0.001) and sVCAM-1 (p < 0.05) were elevated at the day of diagnosis and both parameters reached peak levels three days after onset of chemotherapy. In multivariate analysis soluble adhesion molecules only weakly discriminated between rejection and infection (sensitivity: 13%, specificity: 95%). Although, in combination with routine blood parameters the discriminatory power could be improved (sensitivity: 85%, specificity: 85%) the clinical utility of these markers in non-invasive monitoring is limited.     

Role of beta 1 and beta 2 integrins in the adhesion of human CD34hi stem cells to bone marrow stroma.

Hematopoietic stem cell interaction with elements of the underlying stroma is essential for sustained normal hematopoiesis. Here we have determined that adhesion receptors in the integrin family play a role in promoting adhesion of human hematopoietic stem cells to cultured human marrow stromal cells. Enriched CD34hi progenitor cells expressed VLA-4, VLA-5, and at least one or more beta 2 integrins. Homogeneous marrow stromal cell monolayers capable of supporting proliferation of cocultivated CD34hi cells expressed VCAM-1 and fibronectin (ligands for VLA-4 and VLA-5) as well as ICAM-1 (ligand for LFA-1 and Mac-1). Adhesion-blocking experiments indicated that VLA-4/VCAM-1, VLA-5/fibronectin, and beta 2-integrin/ICAM-1 pathways all are important for CD34hi cell attachment to stromal cells. Consistent with this suggestion, IL-1 stimulation of stromal cells caused both increased VCAM-1 and ICAM-1 expression and increased attachment by CD34hi bone marrow cells. In addition, CD34hi cells utilized VLA-4 to adhere to purified VCAM-1 and employed VLA-5 (and to a lesser extent VLA-4) to adhere to purified fibronectin. Together these results suggest that CD34hi stem cells may utilize multiple integrin-mediated adhesion pathways to localize within specialized microenvironmental niches created by marrow stromal cells.     

Adhesion molecules in untreated inflammatory arthritis: synovial expression and levels in synovial fluid and serum [corrected] [published erratum appears in QJM 1996 Jul;89(7):559]

Adhesion molecules play a critical role in regulating leucocyte migration at sites of inflammation. The relationship of soluble forms in serum or synovial fluid (SF) to synovial membrane expression in inflammatory arthritis is controversial. We examined soluble intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin levels in matched serum and SF, and their relationship to expression in synovium obtained at the same time in 13 patients with previously untreated inflammatory arthritis. Serum- soluble (s)ICAM-1 correlated with sedimentation rate (T = 0.45), Ritchie articular index (T = 0.47) and SF sICAM-1 (T = 0.48), and SF sICAM-1 correlated with membrane ICAM-1 expression (p < 0.02). sE- selectin and sVCAM-1 levels were unrelated to disease activity or membrane expression. Membrane E-selectin expression correlated inversely with ICAM-1 expression (T = -0.57) and serum sICAM-1 (T = - 0.54). Serum sE-selectin correlated inversely with membrane ICAM-1 expression (T = -0.55). The correlations observed between ICAM-1 in serum, SF, synovium and disease activity suggest that ICAM-1 could be a useful target for immunotherapy. The inverse relationship of ICAM-1 and E-selectin suggest important differences in regulation and pathogenetic roles.      

Analysis of T cell stimulation by superantigen plus major histocompatibility complex class II molecules or by CD3 monoclonal antibody: costimulation by purified adhesion ligands VCAM-1, ICAM-1, but not ELAM-1.

Many ligands of adhesion molecules mediate costimulation of T cell activation. The generality of this emerging concept is best determined by using model systems which exploit physiologically relevant ligands. We developed such an "antigen-specific" model system for stimulation of resting CD4+ human T cells using the following purified ligands: (a) major histocompatibility complex class II plus the superantigen Staphylococcus enterotoxin A, to engage the T cell receptor (TCR); (b) adhesion proteins vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), and endothelial leukocyte adhesion molecule 1 (ELAM-1), to provide potential cell surface costimulatory signals; and (c) recombinant interleukin 1 beta (rIL-1 beta)/rIL-6 as costimulatory cytokines. In this biochemically defined system, we find that resting CD4+ T cells require costimulation in order to respond to TCR engagement. This costimulation can be provided by VCAM-1 or ICAM-1; however adhesion alone is not sufficient since ELAM-1 mediates adhesion but not costimulation. The cytokines IL-1 beta and IL-6 by themselves cannot mediate costimulation, but augment the adhesion ligand-mediated costimulation. Direct comparison with the model of TCR/CD3 engagement by CD3 monoclonal antibody demonstrated comparable costimulatory requirements in both systems, thereby authenticating the commonly used CD3 model. The costimulation mediated by the activation-dependent interaction of the VLA-4 and LFA-1 integrins with their respective ligands VCAM-1 and ICAM-1 leads to increased IL-2R alpha (CD25) expression and proliferation in both CD45RA+ CD4+ and CD45RO+ CD4+ T cells. The integrins also regulate the secretion of IL-2, IL-4, and granulocyte/macrophage colony-stimulating factor. In contrast the activation-independent adhesion of CD4+ T cell to ELAM-1 molecules does not lead to T cell stimulation as measured by proliferation, IL-2R alpha expression, or cytokine release. These findings imply that adhesion per se is not sufficient for costimulation, but rather that the costimulation conferred by the VLA-4/VCAM-1 and LFA-1/ICAM-1 interactions reflects specialized accessory functions of these integrin pathways. The new finding that VLA-4/VCAM-1 mediates costimulation adds significance to observations that VCAM-1 is expressed on a unique set of potential antigen-presenting cells in vivo.     

汉防己甲素诱导HL-60细胞分化

目的:探讨汉防己甲素(Tetrandrine)靶向黏蛋白1(MUC1)诱导急性早幼粒细胞白血病(APL)细胞分化。方法:以HL-60细胞株作为研究对象,选取对数生长期细胞,分为空白对照、ATRA、汉防己甲素和ATRA+汉防己甲素,共4组。显微镜下观察各组细胞形态的变化;用NBT还原实验分析细胞分化能力的变化;采用流式细胞术检测细胞表面分化抗原CD11b表达的变化;采用Western blot检测MUC1表达的变化。结果:汉防己甲素可以诱导HL-60细胞分化;ATRA+汉防己甲素组NBT还原阳性率明显高于ATRA和汉防己甲素组(P<0.05);ATRA+汉防己甲素组CD11b+细胞百分率(43.62%±1.38%)明显高于汉防己甲素(15.25%±2.11%)、ATRA(21.84%±7.53%)和空白对照组(8.16%±1.01%)(P<0.05);汉防己甲素组MUC1蛋白量明显低于空白对照组和ATRA组(P<0.05)。结论:汉防己甲素体外能协同ATRA使HL-60细胞分化成熟,其作用机制可能与调控MUC1的表达有关。