促炎症细胞因子刺激人类关节骨膜Ｂ型细胞表达粘附分子

目的：透析相关性淀粉样变（ＤＲＡ）病人关节滑膜组织粘附分子表达增强,并与局部炎与细胞浸润密切相关。旨在探讨ＤＲＡ时秀导滑膜细胞分子表达上调的机制,方法：分离正常人关节滑膜Ｂ型细胞,与晚期糖基化终产物修饰的β微球蛋白（ＡＧＥ－β２m）、天然β２m、肿瘤坏死因子－α（ＴＮＦ－α）,白细胞介素－１β（ＩＬ－１β）在体外共同培养,用荧光单克隆抗体染色,流式细胞仪检测定量分析滑膜Ｂ型细胞表面细胞间粘附分子－１（ＩＣＡＭ－１）,血管细胞间粘附分子－１（ＶＣＡＭ－１）、Ｅ－选择素（Ｅ－selectin)的表达,结果：正常关节滑膜Ｂ型细胞表达ＩＣＡＭ－１、ＶＣＡＭ－１,但不表达Ｅ－selectin.IL-1β、ＴＮＦ－α能以时间和剂量依赖的方式上调滑膜Ｂ型细胞ＩＣＡＭ－１、ＶＣＡＭ－１的表达,但无诱导Ｅ－selectin表达的作用。ＡＧＥ－β２m 和β２m对Ｂ型细胞粘附分子的表达无直接影响,结论：ＤＲＡ时关节组织存在的促炎症细胞因子可能上调滑膜细胞粘附分子的表达,从而促使局部的单核细胞浸润。     

类风湿关节炎滑膜组织及关节液中DcR3表达的研究

目的:观察研究人诱骗受体3(DcR3)在类风湿关节炎(RA)关节液及滑膜组织中的表达及其与滑膜炎性病变程度的相关性.方法:用免疫组织化学的方法对12例RA患者、6例骨关节炎(OA)患者及4例无关节病变的骨折患者滑膜组织中DcR3的表达进行描述分析,并对DcR3表达情况与RA患者滑膜炎性病变程度相关性进行分析.同时用ELISA的方法检测了26例RA及19例OA患者关节液中DcR3的表达.结果:DcR3在RA滑膜组织主要分布于血管翳周围炎性细胞及部分滑膜细胞中,DcR3阳性细胞数约为72％.OA滑膜组织细胞中DcR3也呈现少量的阳性表达,但与RA比较阳性程度较弱,阳性细胞数量较少:对DcR3表达与RA滑膜炎性程度的相关性分析发现,DcR3与RA滑膜炎性程度呈正相关(r=0.832,P＜0.01),同时发现RA患者关节液中表达也明显高于OA患者.结论:DcR3在RA滑膜组织炎性细胞及滑膜细胞上的异常表达可能是其参与RA滑膜炎及滑膜组织侵袭等病理过程,从而导致滑膜损伤的一个重要机制.     

塞来昔布对膝关节炎大鼠肿瘤坏死因子α信号通路及关节滑膜细胞的影响*

目的：探究塞来昔布对膝关节炎大鼠肿瘤坏死因子α（ TNF-α）信号通路及关节滑膜细胞的影响。方法： SD大鼠随机分为模型组、假手术组和治疗组,治疗组大鼠制作膝关节炎模型后给予塞来昔布进行治疗。通过 TUNEL 法、qRT-PCR 法、CCK-8 法、免疫印迹等方法检测3 组大鼠滑膜细胞凋亡情况、TNF-α mRNA表达含量、 TNF-α 蛋白含量、关节炎指数及细胞增殖情况。结果：模型组、治疗组、假手术组关节滑膜细胞凋亡率分别为 89.38%、46.84%、13.68%。模型组与假手术组、治疗组相比较,细胞凋亡率显著升高,治疗组细胞凋亡数量多 于假手术组。假手术组、治疗组细胞增殖OD值比模型组高,差异具有统计学意义,假手术组关节滑膜细胞的增 殖数量最多,模型组关节滑膜细胞的增殖数量最少,治疗组关节滑膜细胞的增殖情况比假手术组相比明显减少。 与假手术组、治疗组比较,模型组关节滑膜细胞中TNF-α 蛋白表达水平显著上升,治疗组TNF-α 蛋白表达水平较 假手术组明显升高。与假手术组、治疗组比较,模型组TNF-α mRNA含量显著升高,治疗组TNF-α mRNA表达 量较假手术组明显升高。治疗组和模型组大鼠关节炎指数相近,随着给药时间的增加,治疗组大鼠膝关节炎情况 逐渐好转,而未服用药物的模型组大鼠病情逐渐恶化。结论：塞来昔布胶囊治疗膝关节炎大鼠效果显著,能够显 著降低TNF-α 信号通路的表达水平,改善关节滑膜细胞的凋亡情况。     

髓样细胞核分化抗原在强直性脊柱炎病人的表达

目的 通过比较强直性脊柱炎(AS)患者和健康志愿者的基因表达谱，寻找与AS相关的新基因并探讨其在病因和发病机制中的意义。方法 用含588个基因的cDNA微阵列，检测AS和健康志愿者的外周血单个核细胞的基因表达语，初步筛选出在AS中差异表达的基因，为了进一步验证cDNA微阵列差异表达基因结果，扩大各组病例数，再用RT-PCR技术同时检测、对比健康志愿者和AS患者外周血单个核细胞(PBMC)、关节液单个核细胞(SFMC)和关节滑膜细胞这些差异表达基因的变化。结果 和健康志愿者组的PBMC比较，AS病人骨髓样细胞核分化抗原(MNDA)、迁移抑制因子相关蛋白8(MRP8)和MRP14、粘附分子ICAM-1和integrin β1表达明显增高，其中MNDA和MRP8的变化相关系数为0．793。在AS的SFMC和关节滑膜细胞中，MNDA表达也显著增高(与健康志愿者组PBMC比较，P值分别为0．038和0．027)；MNDA区别AS和健康志愿者的统计学鉴别准确率达1。AS病人在接受抗TNF-α单克隆抗体治疗3个月后，MNDA在关节滑膜细胞的表达水平显著下降(P＝0．018)。结论 MNDA是AS发病过程中一个重要的并与单核／巨噬细胞致炎密切相关的免疫因子，其可能成为一种用于AS的诊断、治疗监控指标。     

类风湿性关节炎滑膜组织中CD147表达的检测

目的：探讨类风湿性关节炎(RA)滑膜组织CD147的表达。方法：采用免疫组织化学SP染色，双重免疫荧光染色及激光扫描共聚焦显微镜分析检测RA患者受损关节软骨-血管翳汇合部(CPJ)滑膜组织中CD147的表达，并与骨关节炎(OA)滑膜组织CD147的检测相对照。结果：SP染色显示3例OA滑膜组织CD147的表达均为阴性，而11例RA滑膜组织中均检测到CD147的表达，表达CD147的细胞为单核-巨噬细胞、淋巴细胞和滑膜成纤维样细胞。双重免疫荧光染色及激光扫描共聚焦显微镜分析证实表达CD147的细胞为单核-巨噬细胞，中性粒细胞及T细胞。结论：RA滑膜细胞CD147的表达增高。CD147可能是导致RA受损关节软骨、骨基质降解的重要因素之一。     

中药风湿宁对佐剂性关节炎大鼠关节滑膜组织ICAM-1表达的影响

目的：探讨中药复方风湿宁（FSN）N-实验性类风湿性关节炎（RA）的疗效及作用机理。方法：以佐剂性关节炎（adjunvant arthritis,AA）大鼠为实验对象,分别予以FSN与雷公藤多甙（TWP）进行治疗,对照观察大鼠关节肿胀程度,以及关节内病理组织学变化情况。采用免疫组织化学检测各组AA大鼠病变关节滑膜组织中ICAM-1的表达。结果：FSN可显著抑制AA大鼠足跖肿胀,明显减轻病变关节滑膜炎症与增生,防止关节软骨及骨质的破坏;FSN能明显降低ICAM-1在AA大鼠滑膜组织的表达（P〈0.01）结论：FSN具有改善实验性RA症状及病情的作用,其抗炎作用与调控滑膜细胞-细胞间及细胞-细胞外基质间的粘附作用有关。     

PI3K-AKT信号通路及其在类风湿性关节炎滑膜细胞增殖和凋亡中的作用

PI3K-AKT信号通路是重要的细胞内信号转导通路,与类风湿性关节炎(RA)的发生发展密切相关。该信号通路的活化状态受到严密调控:包括负调控因子PTEN、SHIP和正调控因子TNF-α、TGF-β、TRAIL等。RA患者滑膜细胞中此信号通路处于异常激活状态,导致下游抗凋亡基因高表达并且影响下游多种效应分子,在RA患者关节滑膜细胞增殖和凋亡失衡中发挥关键作用。过度增生的滑膜细胞向关节软骨和骨组织浸润生长,导致患者关节畸形和功能障碍。因此,使用PI3K-AKT信号通路抑制剂或上调该信号通路中的内源性负性调节蛋白的表达可逆转RA滑膜细胞的过度增殖,为治疗此疾病提供新靶点。     

骨关节炎患者膝关节滑膜CD1c~+髓源树突细胞的鉴定

CD1c~+髓样树突细胞(myeloid dendritic cells, mDC)是一类重要的DC亚群,参与T细胞的免疫应答。为探讨膝骨关节炎患者关节滑膜是否存在CD1c~+mDC的分布,收集20例骨关节炎(osteoarthritis, OA)患者的外周血和膝关节滑膜,采用FACS检测外周血和关节滑膜内的CD1c~+mDC、单核/巨噬细胞和T细胞,免疫组化方法检测关节滑膜中CD1c、CD3、CD33分子的表达情况。结果显示,OA患者膝关节滑膜中的CD1c~+mDC和巨噬细胞的比例显著高于外周血(P<0.000 1),而T细胞比例显著低于外周血(P<0.000 1);关节滑膜中存在CD1c和CD3的弥漫性浸润且两者定位邻近。该研究提示,OA关节滑膜中存在一群比例显著高于外周血的CD1c~+mDC,CD1c~+mDC在骨关节炎发生、发展过程中具有重要意义。     

敲减骨桥蛋白的膝关节滑膜细胞转录组和蛋白质组定量分析

背景:骨关节炎患者关节软骨和关节液中骨桥蛋白mRNA水平及蛋白表达均较正常人明显增高,但是骨桥蛋白在膝骨关节炎发展进程中的作用及其具体机制在既往文献中尚未阐明。目的:探讨膝骨关节炎患者膝关节滑膜细胞中骨桥蛋白水平及其与骨关节炎严重程度的关系,并通过转录组和蛋白质组测序,进一步阐述其影响骨关节炎进展的机制。方法:选取2019年诊治的42例原发性膝骨关节炎患者,根据胫股关节受累情况,分为单间室骨关节炎组和双间室骨关节炎组。获取滑膜标本及关节液标本,PCR法检测滑膜标本中骨桥蛋白基因在细胞中的表达丰度,ELISA法检测关节液标本中骨桥蛋白水平。应用酶消化法获取培养人膝骨关节炎滑膜成纤维样细胞,用骨桥蛋白siRNA干预24 h后,对样品进行转录组和蛋白质组检测。结果与结论:(1)与单间室骨关节炎组相比,双间室骨关节炎组关节滑膜及关节液中骨桥蛋白均升高,差异有显著性意义(P <0.05);(2)体外培养人膝骨关节炎滑膜成纤维样细胞,通过si RNA敲减骨桥蛋白后,转录组与蛋白质组定量研究分别鉴定了14 662个转录本和3 608个蛋白,其中5种蛋白的转录组与蛋白质组分析结果显示同步下降,2...     

颞下颌关节滑膜A型细胞起源、体外培养、纯化分离及生物学功能的研究进展

颞下颌关节(TMJ)由关节囊包绕,关节囊光滑的内层即为滑膜,各种关节内疾病均会累及滑膜,滑膜是关节疾病的关键性区域。滑膜A型细胞位于滑膜衬里层,多位于滑膜靠近关节腔一侧,具有较强的吞噬作用,主要作用是清除关节腔内和细胞外基质的降解物。目前通过体外细胞培养的方法可以有效地模拟体内细胞的生长环境,并能确切地了解单一和多元因素对滑膜细胞的影响,已成为一种基础研究手段。从细胞起源、体外培养、纯化分离及生物学功能等方面对人TMJ滑膜A型细胞的研究进展进行综述。     

Expression of adhesion molecules in synovia of patients with treatment-resistant lyme arthritis

The expression of adhesion molecules in synovium in patients with Lyme arthritis is surely critical in the control of Borrelia burgdorferi infection but may also have pathologic consequences. For example, molecular mimicry between a dominant T-cell epitope of B. burgdorferi outer surface protein A and an adhesion molecule, human lymphocyte function-associated antigen 1 (LFA-1), has been implicated in the pathogenesis of treatment-resistant Lyme arthritis. Using immunohistochemical methods, we examined synovial samples for expression of adhesion molecules in 29 patients with treatment-resistant Lyme arthritis and in 15 patients with rheumatoid arthritis or chronic inflammatory monoarthritis. In Lyme arthritis synovia, endothelial cells showed intense expression of P-selectin and vascular adhesion protein-1 (VAP-1). Expression of LFA-1 was also intense on infiltrating cells, particularly in lymphoid aggregates, and intercellular adhesion molecule-1 (ICAM-1) was markedly expressed on synovial lining and endothelial and infiltrating cells. Moderate expression of vascular cell adhesion molecule-1 (VCAM-1) was seen on synovial lining and endothelial cells, and mild expression of its ligand, very late antigen-4, was apparent in perivascular lymphoid infiltrates. Except for lesser expression of VCAM-1 in Lyme synovia, the levels of expression of these adhesion molecules were similar in the three patient groups. We conclude that certain adhesion molecules, including ICAM-1 and LFA-1, are expressed intensely in the synovia of patients with Lyme arthritis. Upregulation of LFA-1 on lymphocytes in this lesion may be critical in the pathogenesis of treatment-resistant Lyme arthritis.     

The anti-TNF-α antibody infliximab indirectly regulates PECAM-1 gene expression in two models of in vitro blood cell activation

Chronic inflammatory bowel diseases can be successfully treated with antibodies against the acute phase mediator TNF-α. The process of activation and of extravasation of inflammatory cells from the blood into the 'stressed' tissue site is controlled by cytokines and chemokines, which attract leukocytes and by adhesion molecules, which mediate their attachment and transmigration toward the affected cell(s). The changes in the gene expression of adhesion molecules taking place in those cells before attachment have been less investigated. Changes of PECAM-1, ICAM-1 and vascular cell adhesion molecule-1 (VCAM-1) gene expression were studied in phytohaemagglutinin (PHA)- and lipolysaccharide (LPS)-treated human peripheral blood leukocytes (PBLs), granulocytes and the human monocyte cell line U-937. Cells were treated either with PHA or with LPS in the presence or absence of infliximab and incubated with TNF-α, IFN-γ and/or transforming growth factor beta (TGF-β) and treated as above. Activation of PBLs by PHA or LPS treatment triggered a sharp upregulation of ICAM-1, VCAM-1 gene expression and a time-dependent downregulation of PECAM-1 gene expression reaching a minimum 4?h from start of the experiment. The anti-TNF-α antibody infliximab, by neutralizing TNF-α and IFN-γ production, completely reversed PECAM-1 mRNA downregulation and ICAM-1 and VCAM-1 upregulation. Immunostaining of PBLs cytospins with antibodies against PECAM-1 and ICAM-1 confirmed RT-PCR and western blot results. PBLs IFN-γ or TNF-α treatment downregulated PECAM-1 in parallel with the upregulation of ICAM-1 and VCAM-1 gene expression, whereas TGF-β upregulated PECAM-1- and downregulated ICAM-1 and VCAM-1 gene expression counteracting the effect of TNF-α or IFN-γ. Similar results were obtained in human U937 cells and in granulocyte cultures by TNF-α or IFN-γ treatment. Taken together, these results suggest that infliximab, blocking TNF-α and IFN-γ production, exerts its anti-inflammatory effect through inhibiting downregulation of PECAM-1 gene expression and upregulation of ICAM-1 and VCAM-1 expression in leukocytes of the peripheral blood. These results also suggest that TGF-β may thus be of therapeutic importance as an anti-inflammatory agent.     

Proteasome inhibitors block VCAM-1 and ICAM-1 gene expression in endothelial cells without affecting nuclear translocation of nuclear factor-ϰB

Endothelial cells play a major role in recruiting leukocytes to sites of inflammation. This is accomplished, at least in part, by up-regulation of cell surface adhesion molecules, including VCAM-1 and ICAM-1, in response to cytokines. In this report, we investigated the role of the proteasome complex in mediating the interleukin (IL)-1β induction of VCAM-1 and ICAM-1 gene expression in human endothelial cells. We present evidence that a proteasome inhibitor, n-acetyl-leucinyl-leucinyl-norleucinal (norLEU), as well as specific protease inhibitors, n-tosyl-Lys-chloromethylketone and n-tosyl-Phe-chloromethylketone, blocked IL-1β induction of VCAM-1 and ICAM-1 promoter-driven reporter gene expression in stably transfected endothelial cells. These inhibitors also blocked cytokine induced cell surface expression of VCAM-1 and ICAM-1 by human umbilical vein endothelial cells. As expected, the protease inhibitors blocked the activation of nuclear factor (NF)-?B in response to IL-1β stimulation. In contrast, norLEU did not prevent IL-1β-induced nuclear translocation of NF-?B. The effects of norLEU were specific because it did not inhibit the IL-1β induction of plasminogen activator inhibitor type 1 gene expression. This study demonstrates that inhibition of the proteolytic activity of the proteasome blocks IL-1β induction of VCAM-1 and ICAM-1 gene expression in human endothelial cells.     

Keratinocyte growth factor (KGF) decreases ICAM-1 and VCAM-1 cell expression on bronchial epithelial cells

Activation of leucocytes during airway inflammatory reaction involves adhesion to bronchial epithelial cells (BEC), a process implicating specific interactions between glycoproteins with epithelial cell surface proteins, mainly intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). In this study, the effect of keratinocyte growth factor (KGF), a growth factor involved in pulmonary epithelium repair, was evaluated on adhesion molecule expression with BEAS-2B cells and BEC and granulocyte adherence to BEAS-2B. The modulation by KGF of membrane and mRNA expression of ICAM-1 and VCAM-1 was studied on confluent cells stimulated or not with tumour necrosis factor-alpha (TNF) (200 UI/ml) or TNF and interleukin (IL)-4 (50 UI/ml and 10 ng/ml). Levels of soluble-(s)ICAM-1 and sVCAM-1 were measured by ELISA. Although moderately, KGF significantly decreased membrane ICAM-1 expression in unstimulated BEAS-2B cells (24% inhibition at 100 ng/ml) or in TNF- or TNF + IL-4-stimulated cells (22.5 and 18.7% inhibition, respectively). Treatment with KGF tended to decrease VCAM-1 expression in TNF- and TNF + IL-4-stimulated BEAS-2B (P = n.s. and P < 0.05, 14 and 15% inhibition, respectively). In primary culture of BEC, adhesion molecule expression was also reduced. ICAM-1 and VCAM-1 mRNA expression were also inhibited by KGF. Levels of sICAM-1 and sVCAM-1 were not significantly increased in supernatants from KGF-treated cells (30% and 24% increase at 100 ng/ml, respectively) compared to controls. Moreover, KGF decreased by 31% the adherence of neutrophils to TNF-activated BEAS-2B. In conclusion, KGF decreases ICAM-1 and VCAM-1 expression and neutrophil adherence in BEC. These suggest its involvement in the resolution of the inflammatory reaction.     

Induction of Inflammatory Cytokines in Effusion Cavity by OK-432 Injection Therapy for Patients with Malignant Effusion: Role of Interferon-γ in Enhancement of Surface Expression of ICAM-1 on Tumor Cellsin Vivo

The intracavitary injection of OK-432, a streptococcal preparation, has been shown to be an effective immunotherapy for patients with malignant effusion. We found that this therapy increases surface expression of intercellular adhesion molecule-1 (ICAM-1) on tumor cells, and that the degree of increased ICAM-1 was correlated with therapeutic effects. In the present study, we investigated the ability of OK-432-induced inflammatory cytokines, such as interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β), to enhance ICAM-1 expression. We treated 17 patients who had a malignant effusion with OK-432. At 24 hr after OK-432 injection, ICAM-1 levels on tumor cells were increased significantly in responders except in one case (n= 9), whereas no change was evident in nonresponders except in two cases (n= 8). Induced maximum levels of IFN-γ in responders were significantly higher than those in nonresponders. In contrast, there was no significant difference in the induced TNF-α or IL-1β levels between the two groups. Two types of cultured tumor cells derived from responder patients were successfully established from the 17 different tumor cells in effusion. We performed anin vitrostudy using these cultured tumor cells. Treatment of the two cultured tumor cells with recombinant IFN-γ, but not recombinant TNF-α or IL-1β, significantly increased their ICAM-1 expression to a clinically detectable level. Direct treatment of the tumor cells with cell-free effusion samples obtained from the same patients 24 hr after the therapy successfully increased ICAM-1 expression and this action was blocked completely only by a pretreatment with anti-IFN-γ mAb. Our results suggests that in this therapy IFN-γ plays a crucial role in enhancing ICAM-1 expression by tumor cells and that induced IFN-γ levels may be a useful marker for evaluation of the therapeutic effect.     

CELL ADHESION MOLECULE EXPRESSION WITHIN HUMAN GLOMERULAR AND KIDNEY ORGAN CULTURE

A glomerular and kidney organ culture method was developed to study cytokine inducibility of adhesion molecules and MHC antigen expression. Glomerular cells showed constitutive expression of intercellular cell adhesion molecule-1 (ICAM-1) and MHC class I and II antigens, but not vascular cell adhesion molecule-1 (VCAM-1), E-selectin, or P-selectin. Expression of E-selectin on glomerular endothelial cells (ECs) was induced by interleukin-1 (IL-1), tumour necrosis factor (TNF), interferon-γ (IFN-γ) and granulocyte/macrophage-colony stimulating factor (GM-CSF); this induction was inhibited by transforming growth factor beta (TGFβ) and by IL-4. P-selectin expression was never seen within glomeruli. VCAM-1 was constitutively expressed by Bowman's capsule and proximal tubules and was weakly induced on glomerular ECs by TNF and IL-4 in combination. Glomerular endothelial ICAM-1 expression was increased by IL-1, TNF, IFN-γ, and GM-CSF, while TGFβ inhibited induction by TNF and IL-1. Expression of MHC class I and II antigens by glomerular ECs was constitutive; further upregulation of MHC class II by IFN-γ was observed. These studies suggest that leukocyte–endothelial cell interactions that occur within the kidney follow broadly similar principles as are proposed to occur elsewhere in the body but, in addition, there are subtle differences that reflect local conditions. © 1997 by John Wiley & Sons, Ltd.     

Effect of ultraviolet light on the expression of adhesion molecules and T lymphocyte adhesion to human dermal microvascular endothelial cells

In order to determine the effect of ultraviolet radiation (UVR) on the cell adhesion molecules expressed in human dermal microvascular endothelial cells (HDMEC), the cells were exposed to varying UVR doses and the cell surface was examined for expression of intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM- 1), and E-selectin. The effect of UVB irradiation on the binding of T lymphocytes to HDMEC was also examined. UVA irradiation did not affect the surface expression of ICAM-1, VCAM-1, or E-selectin on the HDMEC. However, following UVB exposure, ELISA demonstrated a significant increase in the baseline ICAM-1 cell surface expression on the HDMEC. However, no induction of either E-selectin or VCAM-1 was noted. UVB also significantly augmented ICAM-1 induction by IL-1alpha and TNF-alpha. VCAM-1 was induced by stimulating HDMEC with IL-1alpha following a UVB irradiation dose of 100 mJ/cm2. Flow cytometric analysis of the HDMEC stimulated with IL-1alpha for 24h demonstrated that 12% of the cells expressed VCAM-1 but either IL-1alpha or UVB irradiation alone failed to induce VCAM-1 expression. Enhancement of T cell-HDMEC binding by IL-1alpha or TNF-alpha treatment was not significantly affected after UVB irradiation. This study demonstrated that UVB irradiation can alter ICAM-1 and VCAM-1 expression on the HDMEC surface and that augmentation of ICAM-1 expression and the IL-1alpha-dependent induction of VCAM-1 following UVB exposure might be important steps in the pathogenesis of sunburn.     

Biological function of CD40 on human endothelial cells: costimulation with CD40 ligand and interleukin-4 selectively induces expression of vascular cell adhesion molecule-1 and P-selectin resulting in preferential adhesion of lymphocytes

The expression of adhesion molecules on vascular endothelial cells determines the pattern of migration and extravasation of leucocytes in inflammation and immunity. Here we show that costimulation with CD40 ligand (CD40L) and interleukin (IL)-4 (or IL-13) gives rise to a unique pattern of adhesion molecule expression by human umbilical vein endothelial cells (HUVEC). CD40 ligation alone enhanced expression of vascular cell adhesion molecule-1 (VCAM-1), intracellular adhesion molecule-1 (ICAM-1) and E-selectin whereas IL-4 and IL-13 increased expression of VCAM-1 and P-selectin but not ICAM-1 or E-selectin. When IL-4 and CD40L were combined there was an additional increase of both VCAM-1 and P-selectin, but ICAM-1 and E-selectin were both inhibited. The combined effects of IL-4 and CD40L signalling were not the result of altered response kinetics, enhanced sensitivity of the endothelium, or increased expression of CD40 or the IL-4 receptor. The rise in VCAM-1 expression induced by combined IL-4 and CD40L stimulation was slower and more sustained than with tumour necrosis factor-alpha (TNF-alpha) and occurred only on a subset (75-80%) of the endothelial cell population compared to 100% with TNF-alpha. Costimulation with IL-4 and CD40L increased adhesion of T cells and B cells above levels obtained with either signal alone, but decreased adhesion of neutrophils. Furthermore, CD40 and IL-4 synergistically increased IL-6 but decreased IL-8 production by HUVEC. These results show that interactions between IL-4 and CD40 on endothelial cells give rise to specific patterns of adhesion molecule expression and cytokine production that may have important implications for lymphocyte and neutrophil migration and function at sites of inflammation.