Introduction: Hepatic diseases comprise inflammations of the liver, which can originate from drug-induced, toxic, autoimmune sources and particularly hepatitis B and C virus infection. The outcome of the disease is li...Introduction: Hepatic diseases comprise inflammations of the liver, which can originate from drug-induced, toxic, autoimmune sources and particularly hepatitis B and C virus infection. The outcome of the disease is linked to interactions between the immune system and the virus, and also depends on the age and immune status of the patient. The aim of this study was to evaluate the association of a MAP3K14 (rs2074292), CD40 (rs1883832) polymorphism and chronic hepatitis B virus carriage in a population from Burkina Faso. Methods: In this case-control analysis, 223 and 173 samples, consisting of 90 and 53 controls and 133 and 120 cases, were examined for MAP3K14 and CD40 respectively. The cases included patients with Chronic Hepatitis B (CHB), cirrhosis or hepatocellular carcinoma (HCC). Genomic DNA extraction was executed using INVITROGEN and FAVORGEN kits. Genotyping of MAP3K14 (rs2074292) and CD40 (rs1883832) gene polymorphisms was accomplished via real-time PCR on the QuantStudioTM 5 Real-Time instrument, followed by allelic discrimination using TaqMan Genotyper Software. Data was interpreted using SPSS version 20 and Epi info version 7.5.2.0. Odds ratios (OR), confidence intervals (CI), and p-values were derived for risk and significance evaluation. Results: This study showed that the heterozygous CT genotype and the mutated T allele of the CD40 (rs1883832) gene are involved in the progression of chronic hepatitis to cirrhosis and hepatocellular carcinoma in HBV-infected patients. However, no association was found between polymorphisms in the MAP3K14 gene (rs2074292) and the progression of HBV infection. By combining the two polymorphisms, we observed either high risk or protection, depending on the genotypes of the MAP3K14 and CD40 genes simultaneously carried by the patient. Conclusion: Polymorphisms of the MAP3K14 and CD40 genes are associated with the evolution of HBV infection.展开更多
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 3...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.展开更多
The 8^th International Workshop on Human Leucocyte Differentiation Antigens （chaired by HZ and managed by BS） was run over a 4-year period and culminated in a conference in December 2004. Here we review the achievem...The 8^th International Workshop on Human Leucocyte Differentiation Antigens （chaired by HZ and managed by BS） was run over a 4-year period and culminated in a conference in December 2004. Here we review the achievements of the HLDA Workshops and provide links to information on CD molecules and antibodies against them, including the 93 new CDs assigned in the 8^th Workshop. We consider what remains to be achieved （including an estimate of the number of leucocyte surface molecules still to be discovered）, and how the field can best move forward.展开更多
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 sp...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.展开更多
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）. Meth...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.展开更多
In order to investigate the expression of CD40 in endothelial cells （ECs） in a variety of injured conditions and the interventional role of Andrographitis Paniculata isolate （API0134）, the thoracic aorta ECs of gu...In order to investigate the expression of CD40 in endothelial cells （ECs） in a variety of injured conditions and the interventional role of Andrographitis Paniculata isolate （API0134）, the thoracic aorta ECs of guinea pigs were cultured in vitro until the third passage, incubated in the presence of media containing xanthine oxidase （XO） and xanthine （Xan） which produced oxygen free radical （OFR group）; oxidized-LDL （ox-LDL group）; XO, Xan and API0134 （OFR＋API0134 group）; or ox-LDL and API0134 （ox-LDL＋API0134 group）. The expression of CD40 in ECs was detected by immunofluorescence assay and reverse transcription-PCR （RT-PCR）. The results showed as compared with the control group, the expression of CD40 in ECs in OFR group and ox-LDL group was increased （P〈0.01）, but attenuated significantly in OFR＋ API0134 group and ox-LDL＋API0134 group （P〈0.05）. It was suggested that mPI0134 could protect atherosclerosis by inhibiting the expression of CD40 molecule in injured ECs.展开更多
AIM： To focus on the role of CD40 and CD40L in their pathogenesis. METHODS： We analyzed by immunohistochemistry the CD40 and CD40L expression in the pouch mucosa of 28 patients who had undergone RPC for UC, in the t...AIM： To focus on the role of CD40 and CD40L in their pathogenesis. METHODS： We analyzed by immunohistochemistry the CD40 and CD40L expression in the pouch mucosa of 28 patients who had undergone RPC for UC, in the terminal ileum of 6 patients with UC and 11 healthy subjects. We also examined by flow cytometry the expression of CEH0 by B lymphoo/tes and monocytes in the peripheral blood of 20 pouch patients, 15 UC patients and 11 healthy controls. RESULTS： Ileal pouch mucosa leukocytes presented a significantly higher expression of CD40 and CD40L as compared to controls. This alteration correlated with pouchitis, but was also present in the healthy pouch and in the terminal ileum of UC patients. CD40 expression of peripheral B lymphocytes was significantly higher in patients with UC and pouch, respect to controls. Increased CD40 levels in blood B cells of pouch patients correlated with the presence of spondyloarthropathy, but not with pouchitis, or inflammatory indices. CONCLUSION： High CD40 expression in the ileal pouch mucosa could be implied in the pathogenesis of pouchitis following proctocolectomy for UC, whereas its increased levels on peripheral blood B lymphocytes are associated with the presence of extraintestinal manifestations.展开更多
Background: Inhibition of the lymphoma surface antigen CD40 by the antagonistic CD40 antibody NVP-HCD122 (HCD122) demonstrates activity in various lymphoma subtypes. In this preclinical in vivo study we examined the s...Background: Inhibition of the lymphoma surface antigen CD40 by the antagonistic CD40 antibody NVP-HCD122 (HCD122) demonstrates activity in various lymphoma subtypes. In this preclinical in vivo study we examined the suitability of positron emission tomography (PET) using the thymidine analogue 3’-deoxy-3’-[18F]fluorothymidine (FLT) for early response assessment upon HCD122 treatment in diffuse large B cell lymphoma (DLBCL). Methods: Immunodeficient mice bearing human DLBCL xenografts (SU-DHL-4) received weekly intraperitoneal injections of HCD122. Tumor growth was followed up until Day 14. Molecular imaging with FLT-PET was performed before (Day 0) and after start of therapy (Day 2 and Day 7). On Day 14 lymphoma xenografts were explanted for immunohistochemical analysis to correlate PET findings with CD40 surface expression on tumor tissue. Results: Treatment with HCD122 significantly delayed tumor growth resulting in a tumor growth inhibition of 45% on Day 14. Significant reduction of tumor-to-background ratio (TBR) of FLT-PET was seen in treated animals on Day 7 and preceded change of tumor volume, thus predicting therapy response to HCD122. Immunohistochemical analysis of xenografts revealed significantly higher CD40 expression on treated than on untreated tissue. Moreover, we found a significant correlation between CD40 expression and FLT-PET response for xenograft tumor treated with HCD122. Conclusions: Treatment of DLBCL with the antagonistic CD40 antibody HCD122 can be monitored with FLT-PET as early as seven days after commencement of therapy and seems to increase CD40 expression on tumor tissue.展开更多
Background: The pathway linking inflammation and thrombosis has been extensively studied. Experimental data support that arterial thrombosis also induces a detectable inflammatory response, which in turn, activates pr...Background: The pathway linking inflammation and thrombosis has been extensively studied. Experimental data support that arterial thrombosis also induces a detectable inflammatory response, which in turn, activates prothrombotic pathways closing a vicious circle that interconnects inflammation and thrombosis. Aim: We designed this study to investigate the causes of inflammatory markers increase after coronary angioplasty. Methods: We analyzed the interrelationship of thrombotic and inflammatory markers and the effect of blocking thrombus formation on the inflammatory response in 50 patients undergoing high thrombotic risk coronary angioplasty. The relationship of platelet number to soluble CD40 Ligand, Interleukin-6 and C-reactive protein blood levels was studied. Half of the study population was treated with standard antithrombotic drugs and the other half with the standard therapy plus platelet GP IIb-IIIa receptor inhibitor Eptifibatide. Results: There was a clear correlation between basal platelet count and sCD40L basal levels, post-angioplasty sCD40L increase and post-angioplasty IL-6 levels and post-angioplasty IL-6 levels with post-angioplasty CRP levels. Postangioplasty CRP, IL-6 and sCD40L blood levels were influenced by GP IIb-IIIa treatment in patients with angiographic thrombus. Conclusion: Platelet aggregation induces a proinflammatory response which is blocked by a GP IIb-IIIa inhibitor agent, particularly in patients with patent angiographic thrombus.展开更多
Toll-like receptors (TLRs) play a key role in B cell-mediated innate and adaptive immunity. It has been shown that interleukin 10 (IL-10)-producing regulatory B cells (B10 cells) can negatively regulate cellular immun...Toll-like receptors (TLRs) play a key role in B cell-mediated innate and adaptive immunity. It has been shown that interleukin 10 (IL-10)-producing regulatory B cells (B10 cells) can negatively regulate cellular immune responses and inflammation in autoimmune diseases. In this study, we determined the effect of TLR4 signaling on the CD40-activated B10 cell competency. The results demonstrated that LPS and CD40L synergistically stimulated proliferation of mouse splenocytes. The percentage of B10 cells in cultured splenocytes was significantly increased after CD40L stimulation but such increase was diminished by the addition of LPS. Such effects by LPS were only observed in cells from WT but not TLR4−/−mice. IL-10 mRNA expression and protein production in B10 cells from cultured splenocytes were significantly up-regulated by CD40L stimulation but were inhibited after the addition of LPS in a TLR4-dependent manner. This study suggests that LPS-induced TLR4 signaling attenuate CD40L-activated regulatory B10 cell competency.展开更多
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 w...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.展开更多
CD40 receptor and its ligand CD40 L are the members of TNFR and TNF family,respectively.CD40 is expressed on antigen-presenting cells(APC)as a monomer,and would be dimerized upon its oligomerization through binding to...CD40 receptor and its ligand CD40 L are the members of TNFR and TNF family,respectively.CD40 is expressed on antigen-presenting cells(APC)as a monomer,and would be dimerized upon its oligomerization through binding to its ligand CD40 L.Such dimer formation was shown to be essential for some CD40 signaling events,including phosphoinositide-3 kinase(PI-3 K)activation,the subsequent B7-2 upregulation and the production of IL-8.CD40 L is primarily expressed on activated T cells and activated platelets,and acts as an important costimulatory molecule especially on T follicular helper cells.It is believed that CD40-CD40 L interaction deeply involved in many immune events for the host defense against pathogens and cancer.CD40-CD40 L interaction mediates intracellular signaling along different pathways,such as the canonical and noncanonical nuclear factorκB pathway,mitogen-activated protein kinases,phosphatidylinositol-3 kinase(PI3K)and the phospholipase Cy pathway,is necessary for successful adaptive immune responses mainly relevant to development of CD8+CTLs,and is deeply involved in cross-talks among T cell,B cell,tumor cell,platelet and etc.However,interaction of CD40 with platelet CD40 L would occur in hemodynamics environments,and the regulation of force on this interaction remain unclear.Besides polymerization of CD40,the hemodynamic environment could be believed to regulate interaction between CD40 and its ligand.To reveal the mechanical regulation mechanism of interaction between CD40 L and CD40,atomic force microscope(AFM)was used in the adhesion frequency assay for measuring two-dimensional(2D)kinetics of CD40 L with CD40(the monomer and its dimer)at zero-force and force-clamp assay for measuring single-bond lifetimes of CD40 L/CD40 complex in a range of forces at single-molecule level.AFM cantilever probes were functionalized by incubation with CD40 L in buffer,and soaked in phosphate-buffered saline(PBS)containing 1%bovine serum albumin(BSA)to block nonspecific binding.Polystyrene dishes were coated with CD40(the monomer and its dimer)and then filled with PBS containing 1%BSA.Our data showed that the adhesion frequency of CD40 L with CD40 monomer and its dimer was 0.10 and0. 15,respectively,showing a higher affinity of CD40 L to CD40 dimer rather than its monomer;the rupture force for CD40 L dissociating from CD40 monomer and its dimer was 23 and 31pN,respectively,demonstrating a higher mechanical strength of complex of CD40 L with CD40 dimer instead of monomer;and increasing tension will prolong first and then shorten the bond lifetime of CD40 L-CD40 complex with force threshold of about 1OpN,and dimerization of CD40 had significantly increased enhanced the bond lifetime of CD40 L-CD40 complex,showing a dimerization-enhanced affinity of CD40 to CD40 L and'Catch-Slip'bond mechanism of CD40 L-CD40 interaction.These results suggest that,through upregulating the binding affinity with CD40 L,the polymerization of CD40 stabilizes the linkers and thereby communication between cell and cell,so does the force being small than the force threshold Both CD40 polymerization-and'Catch bond'-induced enhancement of affinity of CD40 with CD40 L may be necessary for stable cross talks between two cells.This finding may be useful for understanding CD40/CD40 L-induced events importantly in physiological or pathological processes at molecular and cellular level.展开更多
Hyper IgM syndrome is a rare immunodeficiency disease characterized by low or absent IgG, IgA, and IgE levels but normal or elevated level of IgM. It can occur as an acquired form or X linked or autosomal mode of in...Hyper IgM syndrome is a rare immunodeficiency disease characterized by low or absent IgG, IgA, and IgE levels but normal or elevated level of IgM. It can occur as an acquired form or X linked or autosomal mode of inheritance. The X linked form (HIGM1) is a result of mutations in the CD40L ligand (CD40L) gene. We investigate the expression of CD40L on activated T cells from a sporadic male case of HIM with no family history and the B cell function in response to anti CD40L mAb and cytokines. Staining of CD40L on activated T cells from the patient is negative with recently developed anti human CD40L mAb, M90 and M92. The low expression of CD40L on activated T cells from the patient′s mother is also detected. Sequencing of CD40L coding region reveals a 4 bp deletion within the CD40L binding domain. These results indicate that the patient has X linked inheritance pattern (XIGM1). CD40L mAb alone could induce the patient′s PBMCs to secret markedly IgG. Our data suggest that the detection of CD40L expression on ativated T cells may be used to identify sporadic cases of HIM as HIGM1.展开更多
To construct a lentiviral shRNA vector targeting rat CD40 gene and detect its effectiveness of gene silencing in dendritic cells（DCs）, specific siRNA targets with short hairpin frame were designed and synthesized ac...To construct a lentiviral shRNA vector targeting rat CD40 gene and detect its effectiveness of gene silencing in dendritic cells（DCs）, specific siRNA targets with short hairpin frame were designed and synthesized according to the mRNA sequence of rat CD40 gene. DNA oligo was cloned into lentiviral expression vector, and then PCR and sequencing analyses were conducted to verify the constructs. The verified plasmids were transfected into 293T cells that over-express recombinant CD40 in order to select the most effective siRNA targets, shRNA lentiviruses from the selected constructs were propagated and harvested with a virus packaging system, and the virus titers were determined. Western blot and Real-time PCR were performed to determine CD40 expression level in the virusinfected dendritic cells. PCR and sequencing analyses reveal that shRNA plasmids of four targets were successfully constructed. The optimal interfering target was selected, and the virus with a titer of 5 × 10^7 TU/mL was successfully packaged. CD40 expression in rat DCs was knockdown at both mRNA and protein levels by virus infection. In comparison to that of control groups, CD40 mRNA expression and protein expression were decreased by 60.9% and 61.2%, respectively. We have successfully constructed recombinant lentiviral shRNA expression vector targeting rat CD40 gene that can effectively down-regulate CD40 gene expression at mRNA and protein levels in rat DC.展开更多
Objective： To investigate the expression of CD40 and CD40 ligand （CD4OL） on the surface of peripheral blood mononuclear cells（PBMCs） in asthmatic rats and the effect of anti-CD40L McAb on cytokines of PBMCs. Meth...Objective： To investigate the expression of CD40 and CD40 ligand （CD4OL） on the surface of peripheral blood mononuclear cells（PBMCs） in asthmatic rats and the effect of anti-CD40L McAb on cytokines of PBMCs. Methods： Flow cytometry and RT-PCR were used to detect the expression of CD40 and CD40L of PBMCs in asthmatic rats. After the PBMCs was treated with anti-CD40L McAb, ELISA was used to detect the IL-4 and IFN-γ levels of culture supernatants. Results： Compared with the normal control group, the expression of CD40 and CD40L of PBMCs in asthmatic rats increased （P 〈 0.05）. Compared with the untreated group, the level of IL-γ and the ratio of IL-4/IFN-γ decreased after the PBMCs was treated with anti-CD40L McAb（P 〈 0.05）. Conclusion： The expression of CD40 and CD40L on the surface of PBMCs in asthmatic rats was up-regulated. Anti-CD40L McAb can rectify the imbalance of Th1 and Th2 cytokines.展开更多
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 su...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.展开更多
文摘Introduction: Hepatic diseases comprise inflammations of the liver, which can originate from drug-induced, toxic, autoimmune sources and particularly hepatitis B and C virus infection. The outcome of the disease is linked to interactions between the immune system and the virus, and also depends on the age and immune status of the patient. The aim of this study was to evaluate the association of a MAP3K14 (rs2074292), CD40 (rs1883832) polymorphism and chronic hepatitis B virus carriage in a population from Burkina Faso. Methods: In this case-control analysis, 223 and 173 samples, consisting of 90 and 53 controls and 133 and 120 cases, were examined for MAP3K14 and CD40 respectively. The cases included patients with Chronic Hepatitis B (CHB), cirrhosis or hepatocellular carcinoma (HCC). Genomic DNA extraction was executed using INVITROGEN and FAVORGEN kits. Genotyping of MAP3K14 (rs2074292) and CD40 (rs1883832) gene polymorphisms was accomplished via real-time PCR on the QuantStudioTM 5 Real-Time instrument, followed by allelic discrimination using TaqMan Genotyper Software. Data was interpreted using SPSS version 20 and Epi info version 7.5.2.0. Odds ratios (OR), confidence intervals (CI), and p-values were derived for risk and significance evaluation. Results: This study showed that the heterozygous CT genotype and the mutated T allele of the CD40 (rs1883832) gene are involved in the progression of chronic hepatitis to cirrhosis and hepatocellular carcinoma in HBV-infected patients. However, no association was found between polymorphisms in the MAP3K14 gene (rs2074292) and the progression of HBV infection. By combining the two polymorphisms, we observed either high risk or protection, depending on the genotypes of the MAP3K14 and CD40 genes simultaneously carried by the patient. Conclusion: Polymorphisms of the MAP3K14 and CD40 genes are associated with the evolution of HBV infection.
文摘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.
文摘The 8^th International Workshop on Human Leucocyte Differentiation Antigens （chaired by HZ and managed by BS） was run over a 4-year period and culminated in a conference in December 2004. Here we review the achievements of the HLDA Workshops and provide links to information on CD molecules and antibodies against them, including the 93 new CDs assigned in the 8^th Workshop. We consider what remains to be achieved （including an estimate of the number of leucocyte surface molecules still to be discovered）, and how the field can best move forward.
基金Supported by the Public Health Department Foundation of Hunan province, China, No. Y02-42
文摘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.
文摘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.
文摘In order to investigate the expression of CD40 in endothelial cells （ECs） in a variety of injured conditions and the interventional role of Andrographitis Paniculata isolate （API0134）, the thoracic aorta ECs of guinea pigs were cultured in vitro until the third passage, incubated in the presence of media containing xanthine oxidase （XO） and xanthine （Xan） which produced oxygen free radical （OFR group）; oxidized-LDL （ox-LDL group）; XO, Xan and API0134 （OFR＋API0134 group）; or ox-LDL and API0134 （ox-LDL＋API0134 group）. The expression of CD40 in ECs was detected by immunofluorescence assay and reverse transcription-PCR （RT-PCR）. The results showed as compared with the control group, the expression of CD40 in ECs in OFR group and ox-LDL group was increased （P〈0.01）, but attenuated significantly in OFR＋ API0134 group and ox-LDL＋API0134 group （P〈0.05）. It was suggested that mPI0134 could protect atherosclerosis by inhibiting the expression of CD40 molecule in injured ECs.
文摘AIM： To focus on the role of CD40 and CD40L in their pathogenesis. METHODS： We analyzed by immunohistochemistry the CD40 and CD40L expression in the pouch mucosa of 28 patients who had undergone RPC for UC, in the terminal ileum of 6 patients with UC and 11 healthy subjects. We also examined by flow cytometry the expression of CEH0 by B lymphoo/tes and monocytes in the peripheral blood of 20 pouch patients, 15 UC patients and 11 healthy controls. RESULTS： Ileal pouch mucosa leukocytes presented a significantly higher expression of CD40 and CD40L as compared to controls. This alteration correlated with pouchitis, but was also present in the healthy pouch and in the terminal ileum of UC patients. CD40 expression of peripheral B lymphocytes was significantly higher in patients with UC and pouch, respect to controls. Increased CD40 levels in blood B cells of pouch patients correlated with the presence of spondyloarthropathy, but not with pouchitis, or inflammatory indices. CONCLUSION： High CD40 expression in the ileal pouch mucosa could be implied in the pathogenesis of pouchitis following proctocolectomy for UC, whereas its increased levels on peripheral blood B lymphocytes are associated with the presence of extraintestinal manifestations.
文摘Background: Inhibition of the lymphoma surface antigen CD40 by the antagonistic CD40 antibody NVP-HCD122 (HCD122) demonstrates activity in various lymphoma subtypes. In this preclinical in vivo study we examined the suitability of positron emission tomography (PET) using the thymidine analogue 3’-deoxy-3’-[18F]fluorothymidine (FLT) for early response assessment upon HCD122 treatment in diffuse large B cell lymphoma (DLBCL). Methods: Immunodeficient mice bearing human DLBCL xenografts (SU-DHL-4) received weekly intraperitoneal injections of HCD122. Tumor growth was followed up until Day 14. Molecular imaging with FLT-PET was performed before (Day 0) and after start of therapy (Day 2 and Day 7). On Day 14 lymphoma xenografts were explanted for immunohistochemical analysis to correlate PET findings with CD40 surface expression on tumor tissue. Results: Treatment with HCD122 significantly delayed tumor growth resulting in a tumor growth inhibition of 45% on Day 14. Significant reduction of tumor-to-background ratio (TBR) of FLT-PET was seen in treated animals on Day 7 and preceded change of tumor volume, thus predicting therapy response to HCD122. Immunohistochemical analysis of xenografts revealed significantly higher CD40 expression on treated than on untreated tissue. Moreover, we found a significant correlation between CD40 expression and FLT-PET response for xenograft tumor treated with HCD122. Conclusions: Treatment of DLBCL with the antagonistic CD40 antibody HCD122 can be monitored with FLT-PET as early as seven days after commencement of therapy and seems to increase CD40 expression on tumor tissue.
文摘Background: The pathway linking inflammation and thrombosis has been extensively studied. Experimental data support that arterial thrombosis also induces a detectable inflammatory response, which in turn, activates prothrombotic pathways closing a vicious circle that interconnects inflammation and thrombosis. Aim: We designed this study to investigate the causes of inflammatory markers increase after coronary angioplasty. Methods: We analyzed the interrelationship of thrombotic and inflammatory markers and the effect of blocking thrombus formation on the inflammatory response in 50 patients undergoing high thrombotic risk coronary angioplasty. The relationship of platelet number to soluble CD40 Ligand, Interleukin-6 and C-reactive protein blood levels was studied. Half of the study population was treated with standard antithrombotic drugs and the other half with the standard therapy plus platelet GP IIb-IIIa receptor inhibitor Eptifibatide. Results: There was a clear correlation between basal platelet count and sCD40L basal levels, post-angioplasty sCD40L increase and post-angioplasty IL-6 levels and post-angioplasty IL-6 levels with post-angioplasty CRP levels. Postangioplasty CRP, IL-6 and sCD40L blood levels were influenced by GP IIb-IIIa treatment in patients with angiographic thrombus. Conclusion: Platelet aggregation induces a proinflammatory response which is blocked by a GP IIb-IIIa inhibitor agent, particularly in patients with patent angiographic thrombus.
文摘Toll-like receptors (TLRs) play a key role in B cell-mediated innate and adaptive immunity. It has been shown that interleukin 10 (IL-10)-producing regulatory B cells (B10 cells) can negatively regulate cellular immune responses and inflammation in autoimmune diseases. In this study, we determined the effect of TLR4 signaling on the CD40-activated B10 cell competency. The results demonstrated that LPS and CD40L synergistically stimulated proliferation of mouse splenocytes. The percentage of B10 cells in cultured splenocytes was significantly increased after CD40L stimulation but such increase was diminished by the addition of LPS. Such effects by LPS were only observed in cells from WT but not TLR4−/−mice. IL-10 mRNA expression and protein production in B10 cells from cultured splenocytes were significantly up-regulated by CD40L stimulation but were inhibited after the addition of LPS in a TLR4-dependent manner. This study suggests that LPS-induced TLR4 signaling attenuate CD40L-activated regulatory B10 cell competency.
文摘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.
基金supported by the National Natural Science Foundation of China ( 116272109, 11432006)
文摘CD40 receptor and its ligand CD40 L are the members of TNFR and TNF family,respectively.CD40 is expressed on antigen-presenting cells(APC)as a monomer,and would be dimerized upon its oligomerization through binding to its ligand CD40 L.Such dimer formation was shown to be essential for some CD40 signaling events,including phosphoinositide-3 kinase(PI-3 K)activation,the subsequent B7-2 upregulation and the production of IL-8.CD40 L is primarily expressed on activated T cells and activated platelets,and acts as an important costimulatory molecule especially on T follicular helper cells.It is believed that CD40-CD40 L interaction deeply involved in many immune events for the host defense against pathogens and cancer.CD40-CD40 L interaction mediates intracellular signaling along different pathways,such as the canonical and noncanonical nuclear factorκB pathway,mitogen-activated protein kinases,phosphatidylinositol-3 kinase(PI3K)and the phospholipase Cy pathway,is necessary for successful adaptive immune responses mainly relevant to development of CD8+CTLs,and is deeply involved in cross-talks among T cell,B cell,tumor cell,platelet and etc.However,interaction of CD40 with platelet CD40 L would occur in hemodynamics environments,and the regulation of force on this interaction remain unclear.Besides polymerization of CD40,the hemodynamic environment could be believed to regulate interaction between CD40 and its ligand.To reveal the mechanical regulation mechanism of interaction between CD40 L and CD40,atomic force microscope(AFM)was used in the adhesion frequency assay for measuring two-dimensional(2D)kinetics of CD40 L with CD40(the monomer and its dimer)at zero-force and force-clamp assay for measuring single-bond lifetimes of CD40 L/CD40 complex in a range of forces at single-molecule level.AFM cantilever probes were functionalized by incubation with CD40 L in buffer,and soaked in phosphate-buffered saline(PBS)containing 1%bovine serum albumin(BSA)to block nonspecific binding.Polystyrene dishes were coated with CD40(the monomer and its dimer)and then filled with PBS containing 1%BSA.Our data showed that the adhesion frequency of CD40 L with CD40 monomer and its dimer was 0.10 and0. 15,respectively,showing a higher affinity of CD40 L to CD40 dimer rather than its monomer;the rupture force for CD40 L dissociating from CD40 monomer and its dimer was 23 and 31pN,respectively,demonstrating a higher mechanical strength of complex of CD40 L with CD40 dimer instead of monomer;and increasing tension will prolong first and then shorten the bond lifetime of CD40 L-CD40 complex with force threshold of about 1OpN,and dimerization of CD40 had significantly increased enhanced the bond lifetime of CD40 L-CD40 complex,showing a dimerization-enhanced affinity of CD40 to CD40 L and'Catch-Slip'bond mechanism of CD40 L-CD40 interaction.These results suggest that,through upregulating the binding affinity with CD40 L,the polymerization of CD40 stabilizes the linkers and thereby communication between cell and cell,so does the force being small than the force threshold Both CD40 polymerization-and'Catch bond'-induced enhancement of affinity of CD40 with CD40 L may be necessary for stable cross talks between two cells.This finding may be useful for understanding CD40/CD40 L-induced events importantly in physiological or pathological processes at molecular and cellular level.
文摘Hyper IgM syndrome is a rare immunodeficiency disease characterized by low or absent IgG, IgA, and IgE levels but normal or elevated level of IgM. It can occur as an acquired form or X linked or autosomal mode of inheritance. The X linked form (HIGM1) is a result of mutations in the CD40L ligand (CD40L) gene. We investigate the expression of CD40L on activated T cells from a sporadic male case of HIM with no family history and the B cell function in response to anti CD40L mAb and cytokines. Staining of CD40L on activated T cells from the patient is negative with recently developed anti human CD40L mAb, M90 and M92. The low expression of CD40L on activated T cells from the patient′s mother is also detected. Sequencing of CD40L coding region reveals a 4 bp deletion within the CD40L binding domain. These results indicate that the patient has X linked inheritance pattern (XIGM1). CD40L mAb alone could induce the patient′s PBMCs to secret markedly IgG. Our data suggest that the detection of CD40L expression on ativated T cells may be used to identify sporadic cases of HIM as HIGM1.
基金Supported by the National Natural Science Foundation of China(No.30670301)Science and Technology Services of Jilin Province, China(No.20050408-1)+1 种基金PhD Scientific Research Foundation of Ministry of Education of China(No.20050183069)the Jilin Province Talent Development Foundation(No.JRJB2007-2)
文摘To construct a lentiviral shRNA vector targeting rat CD40 gene and detect its effectiveness of gene silencing in dendritic cells（DCs）, specific siRNA targets with short hairpin frame were designed and synthesized according to the mRNA sequence of rat CD40 gene. DNA oligo was cloned into lentiviral expression vector, and then PCR and sequencing analyses were conducted to verify the constructs. The verified plasmids were transfected into 293T cells that over-express recombinant CD40 in order to select the most effective siRNA targets, shRNA lentiviruses from the selected constructs were propagated and harvested with a virus packaging system, and the virus titers were determined. Western blot and Real-time PCR were performed to determine CD40 expression level in the virusinfected dendritic cells. PCR and sequencing analyses reveal that shRNA plasmids of four targets were successfully constructed. The optimal interfering target was selected, and the virus with a titer of 5 × 10^7 TU/mL was successfully packaged. CD40 expression in rat DCs was knockdown at both mRNA and protein levels by virus infection. In comparison to that of control groups, CD40 mRNA expression and protein expression were decreased by 60.9% and 61.2%, respectively. We have successfully constructed recombinant lentiviral shRNA expression vector targeting rat CD40 gene that can effectively down-regulate CD40 gene expression at mRNA and protein levels in rat DC.
文摘Objective： To investigate the expression of CD40 and CD40 ligand （CD4OL） on the surface of peripheral blood mononuclear cells（PBMCs） in asthmatic rats and the effect of anti-CD40L McAb on cytokines of PBMCs. Methods： Flow cytometry and RT-PCR were used to detect the expression of CD40 and CD40L of PBMCs in asthmatic rats. After the PBMCs was treated with anti-CD40L McAb, ELISA was used to detect the IL-4 and IFN-γ levels of culture supernatants. Results： Compared with the normal control group, the expression of CD40 and CD40L of PBMCs in asthmatic rats increased （P 〈 0.05）. Compared with the untreated group, the level of IL-γ and the ratio of IL-4/IFN-γ decreased after the PBMCs was treated with anti-CD40L McAb（P 〈 0.05）. Conclusion： The expression of CD40 and CD40L on the surface of PBMCs in asthmatic rats was up-regulated. Anti-CD40L McAb can rectify the imbalance of Th1 and Th2 cytokines.
文摘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.
