银杏内酯B抑制血小板CD40Ligand表达的分子机制研究

目的探讨银杏内酯B(ginkgolide B)对活化血小板CD40 Ligand(CD40L)的影响以及相关的分子机制。方法取正常人血分离血小板,用不同浓度银杏内酯B孵育血小板5 min,然后用胶原(collagen)刺激血小板活化。用Westernblot分析PI3K表达,Akt磷酸化,CD40L的变化。结果①用collagen刺激血小板聚集,银杏内酯B(0.2、0.4、0.6 g.L-1)预处理血小板5 min,血小板聚集率明显降低,聚集率分别为77%,60%和48%。②Western blot结果显示胶原刺激血小板活化后CD40L表达明显增加,银杏内酯B以剂量依赖方式抑制了CD40L表达。③银杏内酯B对胶原刺激的血小板PI3K表达无明显影响。④collagen刺激血小板活化后Akt的磷酸化增加,银杏内酯B抑制了Akt磷酸化。结论银杏内酯B能够有效抑制collagen诱导的血小板聚集以及CD40L的表达,并明显抑制了Akt磷酸化,表明银杏内酯B能够通过PI3K/Akt信号传导通路抑制血小板活化。

Construction of recombinant eukaryotic expression plasmid containing murine CD40 ligand gene and its expression in H22 cells

AIM: To construct a recombinant murine CD40 ligand (mCD40L) eukaryotic expression vector for gene therapy and target therapy of hepatocellular carcinoma (HCC).METHODS: mCD40L cDNA was synthesized by RT-PCR with the specific primers and directly cloned into T vector to generate middle recombinant. After digestion with restriction endonuclease, the target fragment was subcloned into the multi-clone sites of the eukaryotic vector. The constructed vector was verified by enzyme digestion and sequencing,and the product expressed was detected by RT-PCR and immunofluorescence methods.RESULTS: The full-length mCD40L-cDNA was successfully cloned into the eukaryotic vector through electrophoresis,and mCD40L gene was integrated into the genome of infected H22 cells by RT-PCR. Murine CD40L antigen molecule was observed in the plasma of mCD40L-H22 by indirect immuno-fluorescence staining.CONCLUSION: The recombined mCD40L eukaryotic expression vector can be expressed in H22 cell line. It providesexperimental data for gene therapy and target therapy ofhepatocellular carcinoma.

Effect of Clopidogrel on Platelet Membrane CD40 Ligand in Coronary Artery Disease Patients Undertaking Percutaneous Coronary Intervention

Objectives To investigate the change and clinical significance of clopidogrel on platelet membrane CD40L in coronary artery disease patients before and after percutaneous coronary intervention(PCI). Methods 30 cases who were diagnosis coronary artery diseases(CAD) by coronary angiography, mean age 56±9 years old. All the patients who had no antiplatelet aggregation contraindication, were treated with standard anti angina pectoris drugs. Before PCI, all the patients took clopidogrel 75 mg per day. Activated platelet membrane CD40L express rate was measured by flow cytometry before and after PCI 6 hours. Results Activated platelet membrane CD40L express rate were 3.73±2.15 and 2.46±0.90, respectively in 30 patients before and after PCI 6 hours. Activated platelet membrane CD40L express rate was significantly decrease after PCI 6 hours than that before PCI(P<0.01). Conclusions Clopidogrel has significance effect on platelet membrane CD40L in coronary artery disease patients undergoing PCI. Clopidogrel can suppression platelet activation and prevent thromboembolism event occurrence.

High plasma CD40 ligand level is associated with more advanced stages and worse prognosis in colorectal cancer

BACKGROUND Colorectal cancer(CRC)is often associated with elevated platelet count(>400×10^(9)/L),known as thrombocytosis.The role of CD40 ligand(CD40L),a member of the tumor necrosis factor family,is controversial in CRC.Circulating CD40L is higher in CRC,but its relationship with disease staging and local and distant metastasis is not clear.Although most of the circulating CD40L is produced by platelets,no previous study investigated its relationship with CRC-related thrombocytosis.AIM To investigate the role of CD40L to predict the outcome of CRC and its relation to thrombocytosis.METHODS A total of 106 CRC patients and 50 age and sex-matched control subjects were enrolled for the study.Anamnestic data including comorbidities and histopathological data were collected.Laboratory measurements were performed at the time of CRC diagnosis and 1.5 mo and at least 6 mo after the surgical removal of the tumor.Plasma CD40L and thrombopoietin were measured via enzyme-linked immunosorbent assay,while plasma interleukin-6 was measured via electrochemiluminescence immunoassay.Patient follow-ups were terminated on January 31,2021.RESULTS Plasma CD40L of CRC patients was tendentiously higher,while platelet count(P=0.0479),interleukin-6(P=0.0002),and thrombopoietin(P=0.0024)levels were significantly higher as opposed to the control subjects.Twelve of the 106 CRC patients(11.3%)had thrombocytosis.Significantly higher CD40L was found in the presence of distant metastases(P=0.0055)and/or thrombocytosis(P=0.0294).A connection was found between CD40L and platelet count(P=0.0045),interleukin-6(P=0.0130),and thrombocytosis(P=0.0155).CD40L was constant with the course of CRC,and all baseline differences persisted throughout the whole study.Both pre-and postoperative elevated platelet count,CD40L,and interleukin-6 level were associated with poor overall and disease-specific survival of patients.The negative effect of CD40L and interleukin-6 on patient survival remained even after the stratification by thrombocytosis.CONCLUSION CD40L levels of CRC patients do not change with the course of the disease.The CD40L level is strongly correlated with platelet count,interleukin-6,thrombocytosis,and the presence of distant metastases.

Inhibitory Effect of Clopidogrel on Release of Soluble CD40 Ligand by ADP-activated Platelet in Patients With Non-ST-segment-elevation Acute Coronary Syndromes

Objectives To investigate the inhibitory effect of clopidogrel on release of soluble CD40 ligand （sCD40L） by ADP-activated platelet in patients with non-ST-segment elevation acute coronary syndromes（NSTEACS）. Methods Forty-two patients with NSTEACS were treated with clopidogrel for 6 - 8 days. In order to obtain platelet rich plasma （PRP） samples, the venous blood was drawn before and after treatment, respectively. The platelets were activated by adenosine diphosphate （ADP） , thus releasing sCD40L, sCD40L levels were determined by en- zyme-linked immunosorbent assay （ELISA） at different time of the reaction. Results Plasma sCD40L concentration before treatment was （0. 199 ± 0. 155 ） ng/mL, and （0. 190 ± 0. 176） ng/mL after treatment （ P 〉 0.05 ）. Before treatment the PRP sCD40L level at 20-minute of platelet activation was （4.34 ± 2.51 ） ng/mL, and decreased to （2.79 ±1.93 ） ng/mL after treatment （ P 〈 0. 001 ）. The corresponding level at 40 - minute of platelet activation was （5.29 ± 3. 13 ） ng/mL before treatment and （ 2.87 ± 1.59 ） ng/mL after treatment（ P 〈 0. 001 ）. Conclusions Short-term clopidogrel administration might inhibit the release of sCD40L by ADP-activated platelet in patients with NSTEACS, suggesting that, in addition to its antiplatelet potency, clopidogrel may still have an anti-inflammatory effect.

系统性红斑狼疮患者外周血单个核细胞CD_(40)及CD_(40)配体分子表达的研究

目的 :探讨CD4 0 及CD4 0 配体 (CD4 0L)分子在活跃期SLE患者外周血单个核细胞 (PBMC)上的表达及在SLE发病中的可能作用机制。方法 :采用流式细胞仪检测 13例初发活跃期及 13例复发活跃期SLE患者PBMC表面CD4 0 及CD4 0L分子的表达。结果 :初发及复发活跃期SLE患者PBMC表达CD4 0 分子 (2 5 .94± 10 .10 ) %及 (2 7.34±12 .94) % ,均较正常对照组 (13.99± 5 .2 6 ) %明显升高 (P <0 .0 0 1) ;初发活跃期SLE患者PBMC表达CD4 0L分子 (1.74±1.6 6 ) %较正常对照组 (0 .91± 0 .39) %亦明显升高 (P <0 .0 5 )。初发与复发活跃期SLE患者之间比较PBMC表达CD4 0及CD4 0L分子无显著性差异。结论 :活跃期SLE患者外周血PBMC存在CD4 0 及CD4 0L分子的异常表达 ,提示CD4 0 及CD4 0L分子在SLE患者T、B细胞之间的信号传导及SLE发病机制中可能起着重要的作用。

Expression of CD40 and CD40L on Peripheral Blood Mononuclear Cells in Patients with Condyloma Acuminatum

To observe the expression of CD40/CD40L on peripheral blood mononuclear cells （PBMC） in patients with eondyloma aeuminatum （CA）, flow eytometry was employed to examine the expression of CD40 and CD40L on PMBC in 36 patients with CA and 20 healthy controls. Our results showed that mean level of CD40 expression in CA patients was significantly lower than that in the controls （6.58 %±2.74 % vs 14.81 %±6.12 %, t=5. 703, P〈0.05）; the average level of CD40L in CA patients was also significantly lower than that in the controls （0.73 % ±0.54 % vs 2.67 %±2.43 %, t=3. 532, P〈0.05）. Our resutls suggest that the reduced costimulatory interaction of CD40 and CD40L in CA patients may be one of the important factors responsible for the low cellular immunity.

Role for CD40L in the Therapy of Human Cancer

CD40L-CD40 interaction is central to the control of thymusdependent humoral immunity and cell mediated immune responses. CD40, a member of the tumor necrosis factor receptor （TNF- R） family, has been found on the surface of B lymphocytes, monocytes, hematopoietic progenitors, dendritic cells （DCs）, endothelial cells, epithelial cells and so on. Its natural ligand （CD40 ligand, CD40L）, CD154, a member of the tumor necrosis factor （TNF） family, is mainly expressed on activated CD4^＋ T lymphocytes. A direct growth-inhibitory effect can be found when ligated CD40 is on human breast, ovarian, cervical, bladder, non-small cell lung, and squamous epithelial carcinoma cells. This effect is related to the induction of cell cycle blockage and/or apoptosis. CD40L induces phenotypic and functional maturation of DCs, and can increase tumor immunogenicity through up-regulation of costimulatory molecular expression and cytokine production by epithelial cancer cells. As a result, CD40L can enhance tumor rejection immune responses. Furthermore, by means of a “bystander effect”, even CD40-negative tumor subsets can be eliminated by activated tumor-reactive cytotoxic T lymphocytes（CTL）. This review summarizes recent findings on CD40L recombinant protein and gene therapy-based tumor treatment approaches.

抗人CD154单克隆抗体抑制免疫应答的实验研究

目的 :探讨抗人CD1 54单克隆抗体对免疫应答的抑制作用。方法 :①在体外进行混合淋巴细胞反应 ,研究抗人CD1 54单克隆抗体对淋巴细胞增殖反应的影响 ;②在严重免疫联合缺陷 (SCID)鼠体内重建人的免疫功能后 ,研究抗人CD1 54单克隆抗体在SCID小鼠体内对T细胞增殖的影响和对B细胞功能的影响。结果 :①双向混合淋巴细胞反应结果显示 ,抗体的 5个剂量组的 [3H] -TdR的掺入率均显著低于对照组 (均P <0 .0 1 ) ;②在输注人外周血单个核细胞第 6d、第 1 2d、第 2 1d ,实验组SCID鼠体内人T细胞占SCID鼠淋巴细胞的百分率均明显低于对照组(均P <0 .0 5) ;在输注人外周血单个核细胞后的第 1 2d、第 2 1d、第 31d ,实验组SCID鼠体内人IgG的含量明显低于对照组 (均P <0 .0 5)。结论 :抗人CD1 54单克隆抗体能够在体外抑制淋巴细胞的增殖 。

Soluble CD40 ligand is associated with angiographic severity of coronary artery disease in patients with acute coronary syndrome

Background Recently,studies have disclosed soluble CD40 ligand （sCD40L） during atherosclerosis development and plaque destabilization.The objective of the present study was to test the hypothesis that sCD40L levels are higher in acute coronary syndrome （ACS） patients with a greater extent of angiographic coronary involvement.Methods This cross-sectional study examined ACS patients who underwent coronary angiography by measuring their sCD40L levels.In order to estimate the serum levels of sCD40L,10 ml of peripheral venous blood was drawn within 24 hours of admission.sCD40L levels were measured using an enzyme-linked immunosorbent assay （ELISA,RapidBio,West Hills,CA,USA）.Demographic data,presence of concomitant diseases,ACS characteristics,and angiographic findings were evaluated.A review of medical records and patient interviews were conducted to assess coronary risk factors.And the severity of coronary artery disease was evaluated using the Gensini score index.Results Two hundred and eighty-nine patients were included in the study,of whom 186 were male,with an average age of 64.1±10.0 years.Median sCD40L levels were 1.7 ng/ml （0.3-7.3 ng/ml） and Gensini scores were 50 （0-228）.After adjusting for demographic variables and cardiovascular risk factors,the Gensini score was associated with the natural logarithm of the sCD40L level （Coefficient b=0.002,95％ CI 0.000-0.003,P=0.029）.Conclusion sCD40L levels were independently associated with angiographic severity of coronary artery disease in patients with ACS.

Cationic liposome-mediated transfection of CD40 ligand gene inhibits hepatic tumor growth of hepatocellular carcinoma in mice

Objective: To evaluate the efficacy of cationic liposome-mediated CD40 ligand (CD40L) gene therapy for hepato- cellular carcinoma. Methods: 1×106 of parental H22 cells or H22 cells transfected with the expression vector containing murine CD40L cDNA encoding the entire coding region (pcDNA3.1+-mCD40L) were inoculated subcutaneously into the left flanks of syngenic BALB/C mice. The tumor-bearing mice (tumor nodules 10 mm in maximal diameter) received the treatment of the intratumoral injection of pcDNA3.1+-mCD40L/Transfectam, pcDNA3.1+, or phosphate-buffered saline (PBS), or no treatment. The mice were monitored for tumor growth weekly. We examined mCD40L messenger ribonucleic acid (mRNA) expression by reverse transcription polymerase chain reaction (RT-PCR) and the histologic changes in tumors at two weeks after intratumoral injection using immunohistochemical staining of tumor tissues. Results: All mice inoculated with parental H22 cells developed a tumor subcutaneously, and the tumor size increased progressively within three weeks. However, the mice receiving H22-CD40L cells exhibited complete regression of the tumor two weeks after tumor cell inoculation. The tumor-bearing animals with the treatment of pcDNA3.1+ or PBS, or without treatment had progressive tumor growth, while those mice treated with pcDNA3.1+-mCD40L exhibited a significant inhibition of tumor growth. RT-PCR analysis showed that 783-bp fragments cor- responding to the mCD40L mRNA were amplified only from pcDNA3.1+-mCD40L treated tumors. The tumor samples from pcDNA3.1+-mCD40L-treated mice showed significant lymphocyte infiltration, apoptotic bodies, and confluent necrosis in the tumor tissues. Conclusion: The tumorigenicity of CD40L-expressing cells was abrogated when the cells were implanted subcu- taneously. In vivo gene therapy of established liver tumor nodules in mice by the intratumoral injection of pcDNA3.1+-mCD40L led to significant tumor inhibition. There was mCD40L mRNA expression in the tissues from pcDNA3.1+-mCD40L-treated tumors. The intratumoral injection of pcDNA3.1+-mCD40L induced a strong inflammatory, mainly lymphocytic infiltration of the tumor, and increased the necrotic rate of the neoplastic cells.

Levels of circulating follicular helper T cells,T helper 1 cells,and the prognostic significance of soluble form of CD40 ligand on survival in patients with alcoholic cirrhosis

Background:Excessive drinkers(ED)and patients with alcoholic liver disease(ALD)are several times more susceptible to bacterial and viral infections and have a decrease in antibody responses to vaccinations.Follicular helper T(TFH)cells are essential to select B cells in the germinal center and to produce antibodies.TFH cells express both a membrane-associated and a soluble form of CD40 ligand(sCD40L),in which the latter form is released to circulation upon T cell activation.The effect of alcohol on TFH cells has not been studied.Objectives:The goals of this study are to determine the levels of TFH and T helper 1(Th1)cells in ED and those with alcoholic cirrhosis(AC)when compared to healthy controls and to determine the prognostic significance of sCD40L in a cohort of patients with AC.Methods:Controls,ED,and those with AC were enrolled.Baseline demographic,laboratory tests,and peripheral blood mononuclear cells(PBMCs)were isolated and assessed via flow cytometry for TFH cells.In vitro study was performed to determine the ability of PBMCs to secrete interferon(IFN)-ɣupon stimulation.Serum sCD40L was also determined and its prognostic significance was tested in a cohort of AC patients.Results:The levels of circulating TFH(cTFH)cells were significantly lower in peripheral blood of subjects with ED and AC compared to controls(P<0.05).IFN-ɣsecretion from PBMCs upon stimulation was also lower in ED and those with cirrhosis.Serum sCD40L was significantly lower in ED and AC when compared to that in controls(P<0.0005).Its level was an independent predictor of mortality.Conclusions:Patients with AC had significantly lower level of cTFH and sCD40L.The level of sCD40L was an independent predictor of mortality in these patients.

Growing role of CD40 ligand gene transfer therapy in the management of systemic malignancies besides hepatocellular carcinomas

The article ＂Cationic liposome-mediated transfection of CD40 ligand gene inhibits hepatic tumor growth of hepatocellular carcinoma in mice＂ [doi： 10. 1631/jzus.B0820178] by Jiang et al.（2009） in a recent issue of the Journal of Zhejiang University SCIENCE B was highly thought provoking. The authors have clearly demonstrated the efficacy of CD40 ligand gene therapy in inhibiting the growth of hepatocellular carcinomas. The findings of Jiang et al.（2009） are highly important as they further support and corroborate the rapidly expanding role of CD40 ligand gene therapy in the management of systemic malignancies besides hepatocellular carcinomas.

脑动脉支架置入术后血清炎性标志物与缺血性脑血管事件的关系

目的探讨血清炎性标志物与脑动脉(颈动脉或椎-基底动脉)支架置入术后围手术期缺血性脑血管事件的关系。方法测定44例颈动脉或椎-基底动脉支架置入患者术后1 d的外周血中血清高敏C反应蛋白、基质金属蛋白酶-9及可溶性CD 40配体的水平,同时观察围手术期缺血性脑血管事件的发生情况。结果共有4例出现围手术期缺血性脑血管事件。血清基质金属蛋白酶-9在围手术期出现缺血性脑血管事件者中高于未出现者(1291.16±549.29 ng/ml vs 701.45±497.30 ng/ml,P=0.030),血清高敏C反应蛋白与可溶性CD 40配体在二者之间的差异无统计学意义。结论脑动脉支架置入术后的高基质金属蛋白酶-9水平可能与围手术期缺血性脑血管事件危险增加有一定的相关性。

CD_(40)激发对恶性B淋巴细胞体外生物学行为的影响

目的 观察重组人可溶性CD4 0 配体 (rhsCD4 0 L)及CD4 0 L基因转染细胞 (CD4 0 L TC)对恶性B淋巴细胞体外生物学行为的影响 ,探讨rhsCD4 0 L在肿瘤生物治疗中的可能作用。方法 利用基因工程技术获得了rhsCD4 0 L和CD4 0 L TC ,分别与恶性B淋巴细胞株XG2、XG7、U2 6 6、82 2 6、Raji及Daudi共同作用 ,分析了CD4 0 激发对肿瘤细胞的体外增殖 (锥虫蓝计数法 )、细胞周期 (碘化丙锭掺入法 )、细胞表面共刺激分子的表达 (免疫荧光标记法 )以及细胞凋亡 (Annexin Ⅴ FITC法 )的效应。结果 ①恶性B淋巴细胞株CD4 0 表达呈异质性 ,XG2高表达CD4 0 ,82 6 6、Raji和Daudi中度表达 ,而XG7和U2 6 6不表达CD4 0 。显微镜下观察发现 ,rhsCD4 0 L(5 μg ml)可引起XG2或Daudi细胞的同型聚集 ,该效应在作用 6～ 8h后即可出现 ;与CD4 0 L TC细胞共育后 (肿瘤细胞 :CD4 0 L TC =5∶1) ,XG2 ,Raji和Daudi细胞可粘附于CD4 0 L TC表面 ;②rhsCD4 0 L和CD4 0 L TC均可显著抑制XG2、Raji和Daudi细胞的体外增殖 ,导致XG2细胞呈现G1 期阻滞 ,而Raji和Daudi细胞阻滞于G2 期。并诱导XG2、Raji、Daudi细胞的凋亡 ,凋亡率分别为 :XG2细胞 2 3 3%和 18.8% ,Raji细胞 11.6 %和 8.9% ,Daudi 14 .2 %和 15 .9% ;③表型分析显示

骨髓基质细胞表面CD_(40)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 (CD40 L)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CD3 4 + 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CD40 L的表达 ,并与 5C11(2 0 μg/ml)共同孵育 ,分别于 2 4 ,4 8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 2 4h后 ,收取培养上清 ,与纯化的脐血CD3 4 + 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 (An nexinⅤ )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 ,BMSCIL 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CD3 4 + 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 .0 1)。结论 BMSCCD40 分子配基化可促进IL 6和FL的分泌 ,并支持CD3 4 + 细胞的增殖 ,CD40 /CD40 L可能通过调节细胞因子的释放参与造血调控。

Clinical implications of increased expression of CD40L in patients with acute coronary syndromes

To investigate clinical implications of expression of CD40L in monocytes and changes in serum soluble CD40L in patients with acute coronary syndromes (ACS) Methods Sixteen control and 56 patients, including 24 with stable angina (SA), 20 with unstable angina (UA) and 12 with acute myocardial infarction (AMI) enrolled in this study Expression of CD40L in monocytes was analyzed by flow cytometry and sCD40L levels were measured by ELISA Results Expression of CD40L in monocytes and serum levels of sCD40L in UA and AMI patients were higher than in SA patients and controls In patients with AMI, sCD40L levels showed no significant increase when compared to patients with UA, while AMI patients had a peak level of sCD40L at 24 hours after AMI PTCA induced a marked rise in sCD40L levels in all patients, while CD40L expression in monocytes showed no difference between patients with PTCA, before and after Conclusion Enhanced level of serum sCD40L may be a reliable prognostic indicator for ACS and may represent a marker of coronary disease activity

Distinct Effect of CD40 and TNF-Signaling on the Chemokine/Chemokine Receptor Expression and Function of the Human Monocyte-Derived Dendritic Cells

A key and limiting step in the process of human monocyte-derived dendritic cells （mDCs） for clinical use is their in vitro maturation and in vivo migration. We previously observed that CD40 signal facilitated human mDC growth and maturation. To further explore this process, mDCs generated with GM-CSF and IL-4 were co-cultured with apoptotic tumor cells for 24 hours, followed by incubating with anti-CD40 monoclonal antibody or TNF-a for 48 hours to generate mature DCs. The chemokine/chemokine receptor expression and functions of mature DCs upon various stimuli were determined. The expression of costimulatory molecules on apoptotic tumor cell-loaded mature DCs co-cultured with either anti-CD40 antibody （anti-CD40-DCs） or TNF-a （TNF-DCs） were up-regulated compared to immature DCs, consistent with the abilities of these cytokine to drive DC maturation in vitro. The mRNA levels of chemokines such as stromal cell-derived factor-1a （SDF-1a）, EBV-induced molecule 1 ligand chemokine （ELC）, and IFN inducible protein-10 （IP-10） in anti-CD40 activated DCs were increased and the dendritic cell-specific chemokine 1 （DC-CK1） was moderately up-regulated as compared with other mature DCs. The corresponding chemokine receptors CXCR4 and CCR7 of anti-CD40-DCs were significantly expressed. The CXCR3 expression on activated T cells stimulated by anti-CD40-DCs was also increased. Moreover, the anti-CD40-DCs had a stronger ability to stimulate T cell proliferation than any other DCs. The NF-xB activity was much higher in anti-CD40-DCs than that of TNF-DCs. These results offer further evidence of the importance of the CD40 signal in developing efficient human DC vaccines for cancer immune therapy. Cellular ＆ Molecular Immunology.

CD40/CD40L Dyad in the Inflammatory and Immune Responses in the Central Nervous System

CD40 and its cognate ligand （CD40L） are a pair of regulators of pro-inflammatory and immune responses. In the central nervous system （CNS）, CD40 is expressed on a variety of cells, including vascular endothelial cells, smooth muscle cells, astrocytes and microglia （the brain macrophages, being the most sensitive cell type to respond to CD40 ligand）. Interaction between CD40 on microglia and CD40L presented by infiltrating T lymphocytes and other resident CNS cells triggers a series of intracellular signaling events that promote the production of a wide array of cytokines, chemokines and neurotoxins. Thus, both molecules serve as amplifiers of pro-inflammatory and immune responses in the CNS and constitute important molecular targets for therapeutic intervention of diseases.Cellular ＆ Molecular Immunology. 2006;3（3）：163-169.

Role of platelet TLR4 expression in pathogensis of septic thrombocytopenia

BACKGROUND： Infection-induced thrombocytopenia （TCP） is an independent risk factor for death of patients with sepsis, but its mechanism is unknown. This study aimed to explore the underlying mechanism of TCP based on the relationship between TLR4 expression and platelet activation in septic patients. METHODS： A total of 64 patients with sepsis were prospectively studied. Platelet count （PC）, mean platelet volume （MPV）, platelet distribution width （PDW）, platelet TLR4 expression, platelet PAC-1 expression, sCD40L and TNF-a concentrations were compared between the healthy control group （15 volunteers） and sepsis group （64 patients） at admission and on the 3, 5, and 9 days after admission. The changes of MPV and PDW in the TCP and non-TCP subgroups of sepsis before and after treatment were recorded. Prognostic index was analyzed. RESULTS：PC was lower in the sepsis group （P=0.006）, and MPV and PDW were higher in the sepsis group than those in the healthy control group （P=0.046, P=0.001）. Platelet TLR4 and PAC-1 expressions, and sCD40L and TNF-a levels increased more significantly in the sepsis group （P〈0.001）. PAC-1 expression and TNF-a level were higher in the TCP group than in the non-TCP group before and after treatment （P=0.023, P=0.011）. sCD40L concentration and platelet TLR4 expression were significantly higher in the treated TCP group than in the non-TCP group （P=0.047, P=0.001）. Compared to the non-TCP group, the rate of bleeding was higher （P=0.024） and the length of ICU stay was longer （P=0.013）. The APACHE II score and the 28-day mortality were higher in the TCP group （P〈0.01, P=0.048）. CONCLUSIONS：The elevation of platelet TLR4 expression in sepsis along with platelet activation is closely related to the incidence of thrombocytopenia. The occurrence of TCP is a sign of poor prognosis in sepsis patients.