rshCD_（40）L、IFN-γ对食管癌Eca109、Eca9706、TE13细胞增殖和凋亡的影响

目的观察可溶性重组人CD40L（rshCD40L）、IFN-γ对食管癌Eca109、Eca 9706、TE13细胞增殖和凋亡的影响。方法取正常培养的食管鳞癌细胞株Eca109、Eca 9706、TE13,分别用PBS、100 U/ml IFN-γ、100 ng/ml rsh-CD40L、100 U/ml IFN-γ＋100 ng/ml rshCD40L培养,分别为A、B、C、D组。用MTT法测算各组细胞增殖抑制率,用TUNEL法检测细胞凋亡率。结果 C组Eca109、Eca9706、TE13细胞增殖抑制率分别为40.6%±4.2%、31.5%±5.7%、44.6%±6.7%,明显高于A、B组（P均〈0.05）;D组分别为56.7%±4.9%、41.2%±6.2%、51.6%±5.2%,均高于C组（P均〈0.05）。C组Eca109、Eca9706、TE13细胞凋亡率分别为33.6%±3.7%、30.5%±2.8%和37.6%±4.9%,明显高于A、B组（P均〈0.05）;D组分别为43.7%±4.7%、34.2%±5.1%、41.5%±5.7%,均高于C组（P均〈0.05）。结论 rshCD40L能促进食管癌Eca109、Eca9706、TE13细胞凋亡,并抑制其增殖。IFN-γ可增强这一作用。     

Soluble CD40L, platelet surface CD40L and total platelet CD40L in congestive heart failure: relationship to platelet volume, mass and granularity

Background. Many complications associated with congestive heart failure (CHF) have a thrombosis-related aetiology, which may involve platelets. The immune modulator pair CD40-CD40L has been proposed to be an important link between inflammation and thrombosis and may be important in the pathophysiology of CHF. Objective. To study soluble CD40L (sCD40L), platelet surface CD40L (%GCD40L) and total platelet CD40L (pCD40L) levels in CHF patients, their relationships to other platelet indices (platelet volume, mass and component) and to assess their prognostic value. Methods. We measured sCD40L (by ELISA); pCD40L (by a platelet lysate assay); platelet surface CD40L (%GCD40L) expression by flow cytometry; mean platelet volume (MPV), mean platelet mass (MPM) and mean platelet component (MPC); in 108 patients with stable CHF. Levels were compared with 37 ‘healthy controls’ and 63 ‘disease controls’. After a median follow-up period of 490 days, clinical endpoints were determined. Results. pCD40L was significantly higher in CHF than disease controls, but not sCD40L or %GCD40L levels. CHF patients and disease controls had higher MPV (one-way anova , P < 0.0001), whilst MPC was significantly lower in CHF patients (P < 0.0001), compared to healthy controls. All indices related to CD40L (i.e. sCD40L, pCD40L and %GCD40L) were neither related to disease severity or left ventricular ejection function, nor to clinical endpoints at follow-ups. Conclusion. Patients with stable CHF patients did not exhibit enhanced levels of CD40L and the latter did not predict clinical events at follow-up. The lack of difference in CD40L levels between CHF and disease controls suggests that CD40L may not have a major role in CHF per se, but in the comorbidities associated with CHF.     

高血压并发症与患者血清可溶性CD40及CD40L水平的关系

目的探讨原发性高血压患者可溶性CD40/CD40L水平变化。方法根据WHO关于高血压器官损伤并发症的分类,将研究对象分为无并发症组、中度器官损伤及重度器官损伤组。应用酶联免疫反应法测定328名研究对象血清可溶性CD40（sCD40）及可溶性CD40L（sCD40L）水平。结果①与无并发症组（n=180）相比,sCD40水平在重度器官损伤组（n=50）显著升高（39.9±2.6vs30.0±1.0）ng/m l,P<0.01;与中度器官损伤组（29.4±1.4）ng/m l,（n=98）比较,重度器官损伤组sCD40水平也显著升高（P<0.01）;②与无并发症组相比,重度器官损伤组中血清sCD40水平仅在中风组上调;③sCD40L在各组均无显著变化。结论血清sCD40/sCD40L水平与高血压并发症的严重程度有关。     

CD40/CD40L在毛细支气管炎发病机制中的作用

CD40/CD40L是T、B细胞表达的重要膜表面蛋白质分子,它们之间的相互作用对T、B细胞具有重要的影响,在炎症反应、免疫紊乱等方面具有重要的作用.它们之间的交联反应促进气道炎症反应和气道高反应性.目前认为CD40/CD40L在毛细支气管炎发病的多个环节中发挥作用,本文就其与毛细支气管炎的关系进行综述.     

Inverse correlation between soluble CD40 ligand and soluble CD40 is absent in patients with unstable angina

The CD40/CD40 ligand (CD40L) system mediates inflammatory processes important in atherogenesis and plaque instability. The expression of CD40L on activated T cells was suppressed by soluble CD40 (sCD40) in vitro. However, the relationship between soluble CD40L (sCD40L) and sCD40 in unstable angina (UA) is still unknown. Thirty-seven consecutive patients with recent chest pain or discomfort were recruited. Patients with both Braunwald's class IB–IIIB and with coronary stenosis (or stenoses) of >75% were assigned to the UA group (n = 19, aged 67.2 ± 8.2 years), and the rest to the control group (n = 18, aged 63.4 ± 8.7 years). The serum levels of sCD40L and sCD40, and the plasma levels of matrix metalloproteinase (MMP)-9, were measured by enzyme-linked immunosorbent assays. A significantly inverse correlation between sCD40L and sCD40 was shown in the controls (r = −0.72, P = 0.0007), but was absent in the UA group (r = −0.16, P not significant), although there was no statistical significance between these groups in terms of serum levels of sCD40L or sCD40. The difference of the regression slopes of these regression lines was statistically significant (P < 0.01). Additionally, there was a significant correlation between sCD40 and plasma levels of MMP-9 in the patients with and without UA (r = 0.58, P = 0.0096), but no significant correlation between sCD40L and MMP-9 levels (r = 0.00, P not significant). The balance between CD40 and CD40L may be lost in patients with UA. Soluble CD40 expression may also be related to MMP-9 expression in atherosclerotic tissues.     

Coexpression of CD40 and CD40 ligand in B-cell lymphoma cells

CD40 ligand (CD40L) is involved in the T-cell-dependent regulation of B-cell growth and survival and can rescue normal germinal centre B cells and several types of malignant B cells from apoptosis in vitro . We have previously reported that serum of patients with chronic lymphocytic leukaemia contained elevated levels of biologically active soluble CD40L (sCD40L). Whether an augmented CD40L pathway exists in patients with other types of B-cell lymphoid malignancies and the source of native sCD40L in these patients is currently unknown. Using a sensitive ELISA assay, soluble CD40L (sCD40L) was detected in the sera of both healthy individuals and patients with haematological malignancies; however, its level was significantly elevated only in patients with B-cell lymphomas ( P   < 0.0001). Several types of malignant B cells coexpressed CD40 and CD40L proteins, and CD40L mRNA was detected in purified resting malignant B cells. The dual expression of CD40 and CD40L in B cells and the presence of native sCD40L in human serum suggest that a direct T–B-cell contact may not be required for CD40L delivery to B cells. This data raises the possibility that an autocrine cytokine loop involving CD40L may contribute to the growth regulation of benign and malignant B cells in vivo .     

Elevated levels of biologically active soluble CD40 ligand in the serum of patients with chronic lymphocytic leukaemia

Chronic lymphocytic leukaemia (CLL) is an indolent lymphoproliferative disorder manifested by low growth fraction and prolonged survival of the malignant cells. The mechanisms that enable CLL cells to live longer and to resist apoptosis remain unclear. Because the malignant CLL cells express CD40 and Fas receptors, which can transduce cell-survival and cell-death signals, we examined the role of CD40 in the growth regulation of CLL cells and its interaction with Fas-mediated and fludarabine-induced apoptosis in vitro . Primary CLL cells underwent spontaneous apoptosis in culture, which was enhanced by exogenous human Fas ligand (FasL) or fludarabine. Exogenous CD40L rescued CLL cells from spontaneous apoptosis in a dose-dependent manner, and caused CLL cells to resist apoptosis induced by FasL or fludarabine. Patients' autologous plasma rescued CLL cells from spontaneous apoptosis, an effect that could be reversed with anti-CD40 ligand (CD40L) antibodies. The levels of soluble CD40 ligand in the sera of 51 CLL patients and 55 healthy donors were determined by enzyme-linked immunosorbent assay. The mean soluble CD40L level in normal donors was 0.29 ng/ml compared to a mean value of 0.80 ng/ml in CLL patients ( P  < 0.001). CD40L up-regulated bcl-X_L mRNA but not bcl-2 in CLL cells within 3–6 h in culture. Our results demonstrated that serum of patients with CLL contained elevated levels of biologically active soluble CD40L, and that CD40L can prolong survival of CLL cells and mediate their resistance to FasL and fludarabine in vitro .     

HIV感染者/AIDS患者外周血CD38 HLA-DR分子在CD4+ CD8+T淋巴细胞上的表达

目的　观察国内艾滋病病毒 (HIV)感染者 /艾滋病 (AIDS)患者外周血CD38、HLA DR分子在CD+4 、CD+8T淋巴细胞上表达的变化 ,并探讨这些变化的临床意义。方法　用流式细胞仪检测 5 1例正常对照、14例HIV感染者和 3 6例AIDS患者的外周血CD+4 、CD+8T淋巴细胞表面的CD38、HLA DR分子的表达 ,用分枝DNA(bDNA)法检测 11例HIV感染者和 18例AIDS患者的血浆病毒载量。结果　CD+4 HLA DR+细胞百分比显示 ,AIDS组显著高于正常组及HIV组 ;CD+8HLA DR+T细胞百分比显示HIV组与AIDS组间无差异 ,而它们均显著高于正常组。CD+8、CD38+细胞百分比则是AIDS组 >HIV组 >正常组 ,CD+8CD38+、CD+8HLA DR+、CD+4 HLA DR+细胞百分比与病毒载量显著正相关。结论　在HIV感染过程中 ,HLA -DR+、CD38+在CD+4 、CD+8T淋巴细胞上的表达均显著增加 ,反映T淋巴细胞异常激活 ;尤其是CD+8CD38+细胞百分比随着疾病进展逐渐升高 ,预示疾病进展程度。在评价HIV感染者和AIDS患者的免疫状况时 ,不仅要考虑免疫细胞数量和功能的变化 ,还应考虑免疫细胞的激活水平     

共刺激分子CD40/CD40L及B7/CD28与动脉粥样硬化

共刺激分子CD40/CD40L及B7/CD28是T细胞活化的两条重要辅助刺激通路,不仅在调节T细胞免疫反应中起关键作用,而且也在B细胞的活化、增殖、分化、抗体的分泌起重要作用.近年来它们在动脉粥样硬化方面的研究成为国内外学者研究的热点.对共刺激分子和动脉粥样硬化关系的深入了解,将有助于阐明动脉粥样硬化发生的炎症和免疫机制,并为临床治疗开辟新途径.本文综述了共刺激分子的免疫学特征及功能以及在动脉粥样硬化中发挥的作用与免疫学治疗的前景.     

CD40-CD40L在动脉粥样硬化和血栓形成中的作用

近年来研究证实CD40分子与其配体CD40L在动脉粥样硬化的各个阶段均起重要作用。动脉粥样硬化的关键细胞成分——内皮细胞、巨噬细胞和平滑肌细胞上均有CD40和CD40L表达,两者结合能诱导人血管内皮细胞表达多种活性介质,参与动脉粥样硬化斑块的形成,而阻断CD40- CD40L通路可防止动脉粥样硬化或阻止已形成的斑块进展。CD40L可能参与血栓形成和血小板活化,急性脑梗死和急性冠状动脉综合征患者血液中的可溶性CD40L持续性增高。一些药物能够下调CS40L水平,为预防血管事件的发生提供了一个新的途径。     

CD40-CD40L系统与易损斑块的破裂

易损斑块的破裂是急性心血管事件发生的罪魁祸首，CD40-CD40L系统作为免疫和炎症反应的枢纽，其贯穿急性冠脉综合症（ACS）发生发展至斑块破裂及血栓形成的全过程，阻断其信号传导途径可起到一定的预防效果，提示在临床上可作为一种新的治疗途径。     

CD40-CD40L及基质蛋白酶在大鼠动脉粥样硬化中的表达

目的：为探讨粘附分子CD40、CD40配体（CD40 Ligand，CD40L）及基质蛋白酶在动脉粥样硬化中的表达；方法：利用高脂饮食制作大鼠动脉粥样硬化模型（AS组，n=6），与正常饮食大鼠模型（N组，n=6）作对照，流式细胞技术法检测外周血中CD40及CD40L，Zymography法检测外周血中基质蛋白酶-2（MMP-2）及基质蛋白酶-9（MMP-9）的表达情况，免疫组织化学法观察CD40、CD40L及MMP-2、MMP-9在主动脉弓表达情况；并对CD40、CD40L与MMP-2、MMP-9做相关性分析；结果：动脉粥样硬化模型大鼠外周血CD40、CD40L、MMP-2、MMP-9表达均高于正常饮食组大鼠，免疫组织化学法显示CD40、CD40L在主动脉弓内膜上表达，MMP-2、MMP-9在主动脉弓内皮细胞及内皮细胞间质中表达，而且AS组表达强于N组；CD40、CD40L与MMP-2、MMP-9有相关性；结论：提示动脉粥样硬化的形成可能与CD40、CD40L及基质蛋白酶的异常表达有关。     

阿司匹林、肝素对血小板活化及表达CD40L的影响

目的探讨血小板活化与表达CD40L的关系以及阿司匹林、肝素对此过程的影响.方法体外分离健康人血小板,经不同浓度二磷酸腺苷(ADP)、凝血酶诱导后,应用流式细胞术测定血小板活化指标P选择素和炎性标志CD40L表达水平,观察二者随诱导时间延长的变化过程,并分析阿司匹林、普通肝素、低分子肝素对血小板活化及表达CD40L的影响.结果 ADP、凝血酶均呈浓度依赖方式增加血小板P选择素、CD40L表达,二者表达水平随诱导时间延长而同步增减,呈显著正相关(P<0.05).阿司匹林(2.5 μg/ml)对ADP(4 μmol/L)和凝血酶(1U/ml)诱导的血小板活化及CD40L表达无任何影响(P>0.05);普通肝素(2.5U/ml)和低分子肝素(2.5U/ml)单独或与阿司匹林合用,均能极显著地抑制凝血酶诱导的血小板活化及CD40L表达(P＜0.001),但对ADP的诱导过程无影响(P>0.05).结论炎性介质CD40L可表达在活化血小板表面,在多种诱导剂存在的情况下,阿司匹林及肝素能部分抑制血小板的活化及CD40L表达.     

Enhanced plasma soluble CD40 ligand levels in essential hypertensive patients with blunted nocturnal blood pressure decrease

BACKGROUND: Hypertensives with a blunted nocturnal blood pressure (BP) decrease have increased risk of developing atherosclerotic disease. Soluble CD40 ligand (sCD40L) is involved in the pathogenesis of risk factor-related vascular damage. Therefore, we evaluated the relationship between circulating sCD40L levels, circadian BP profile, and early carotid atherosclerosis in essential hypertensives. METHODS: Plasma sCD40L concentrations were assessed in two groups of 25 never-treated hypertensives, without additional cardiovascular risk factors, differentiated on the basis of a nocturnal decrease of BP either of >10% (dippers) or <10% (nondippers) of daytime values, and in 25 matched normotensives. Carotid intima-media thickness (IMT) was also measured in all participants. RESULTS: Plasma sCD40L concentrations were higher in nondippers (4.9 +/- 1.2 ng/mL) than in dippers (3.7 +/- 0.7, P = .0005) and controls (1.6 +/- 0.6, P < .0001). These latter had lower sCD40L concentrations than dippers (P < .0001). The IMT was higher in both hypertensive groups than in normotensives (P < .0001). In the entire hypertensive population IMT directly correlated with circulating levels of sCD40L (r = 0.365, P = .01) and inversely correlated with nocturnal systolic BP decreases (r = -0.286, P = .043). In a multivariate regression analysis sCD40L was the main determinant of IMT (r(2) = 0.157, P = .004). CONCLUSIONS: Nondippers have enhanced plasma sCD40L levels, which may contribute to their increased susceptibility to develop vascular damage.     

Significance of chemokines and soluble CD40 ligand in patients with autoimmune thrombocytopenic purpura

We investigated the levels of various chemokines and soluble CD40L (sCD40L) in ITP patients, in order to determine the influence of CD40-CD40L interaction on the pathogenesis of ITP. We found increases in MCP-1 and RANTES levels in ITP patients compared with those in healthy individuals. Thirty-eight of the 65 ITP patients (58.5%) had elevated levels of sCD40L. We found significant decreases in platelet counts in sCD40L-positive ITP patients. Although the sCD40L level did not differ significantly between the control and nonimmune thrombocytopenia groups, but among ITP patients. sCD40L level was significantly higher in those with untreated ITP than in those with treated ITP. In addition, significant increases in RANTES, MCP-1, sCD14, and sP-selectin levels were observed in sCD40L-positive ITP patients, although sE-selectin levels were not increased in such patients. For other factors examined, however, there were no differences in level between sCD40L-positive and -negative ITP patients. These findings suggests that there are two groups of ITP patients, one with elevated and one with normal of sCD40L. ITP cases in which sCD40L was increased appeared to involve changes in platelet counts and monocyte activation. The pathogenesis of ITP may in some patients include alterations of the CD40/CD40L pathway.     

CD5+B细胞与扩张型心肌病相关性研究

目的:通过对扩张型心肌病(DCM)患者体内CD5 B细胞及其功能的检测,探讨它在DCM发生发展中的作用。方法:选择DCM患者42例,20例健康者为对照。采用双色流式细胞术检测患者外周血淋巴细胞中CD5 B细胞(CD19 CD5 )数,ELISA法检测血清抗心肌自身抗体(AHA)以及体外CD5 B细胞分泌IL-10功能;根据彩色多普勒检测心脏射血分数(EF)。结果:与对照组[(5.39±1.46)%]相比,DCM患者CD5 B细胞均有明显增多,其中AHA阳性者为(13.49±3.15)%,AHA阴性者(14.06±5.23)%,AHA阳性和阴性者CD5 B细胞差异无统计学意义。对照组AHA检测均为阴性。DCM患者CD5 B细胞的增多与心脏EF值无相关性(r=-0.25,P>0.05),但其分泌IL-10功能与EF值负相关(r=-0.64,P<0.01)。结论:CD5 B细胞不仅参与AHA阳性也参与AHA阴性DCM发病过程;有活性的CD5 B细胞介导DCM心力衰竭的进程。     

单核细胞和血小板CD40L高表达及血小板CD62P高含量与急性冠脉综合征的关系

目的探讨急性冠脉综合征(ACS)外周血单核细胞和血小板表达CD40L及血小板CD62P含量的变化及意义。方法应用间接免疫荧光流式细胞仪测定患者与对照组血单核细胞和血小板表达CD40L水平及血小板CD62P含量。结果发现急性冠脉综合征患者(ACS)单核细胞和血小板表达CD40水平及血小板CD62P含量均显著高于正常对照者(NS)(P<0.01)和稳定型冠心病患者(SA)(P<0.05或P<0.01)。结论急性冠脉综合征患者外周血单核细胞和血小板表达CD40L及血小板CD62P含量可能与ACS的发生有关,是动脉粥样硬化斑块不稳定的标志。     

炎性因子CD40配体在急性冠脉综合征患者中的作用

目的探讨急性冠脉综合征（ACS）患者血清可溶性CD40配体（sCD40L）和超敏C反应蛋白（hs-CRP）的水平及临床意义。方法按照诊断标准入选研究对象137例,分为两组：冠状动脉造影无异常者为对照组,21例,ACS组116例,男86例,30例,年龄35～77（56.9±12.6）岁,包括不稳定型心绞痛88例,急性心肌梗死28例。采集患者空腹肘静脉血,采用酶联免疫吸附（ELISA）方法测定血清sCD40L浓度和hs-CRP浓度。所有患者入院第2～4天行冠状动脉造影检查,用直径法测定冠状动脉狭窄的程度,用Gensini评分系统进行评分,累计积分。结果 ACS组sCD40L与hs-CRP水平均高于对照组（P＜0.05）。sCD40L水平与Gensini积分呈正相关关系（r＝0.328,P＝0.000）,hs-CRP水平与Gensini积分呈正相关关系（r＝0.748,P＝0.000）,sCD40L与hs-CRP之间呈正相关关系（r＝0.192,P＝0.039）。结论 ACS患者外周血清sCD40L和hs-CRP水平明显升高,提示CD40/CD40L系统与ACS的发生有关。