Research progress on OX40/OX40L axis mediated T cell immunity in the course of oral lichen planus / 口腔疾病防治

@#Oral lichen planus (OLP) is a common chronic disease of the oral mucosa with unclear pathogenesis. Local infiltration of T cells plays a key role in the pathological process of OLP. Increased evidence supports the notion that the imbalance of helper T cells (Th) 1/Th2 and Th17/regulatory T cells (Treg) and their related cytokines is closely related to the pathogenesis and progression of OLP. In recent years, studies have shown that OX40 (CD134) and its ligand OX40L (CD252) play an important role in the process of the T-cell immune response. They participate in the balance regulation of Th1/Th2 and Th17/Treg, mediate the imbalance of pro-inflammatory and anti-inflammatory, and affect the occurrence and development of a variety of autoimmune diseases. However, there is no direct evidence that the OX40/OX40L axis mediates the imbalance of T-cell subsets in the pathogenesis of OLP. Therefore, large sample clinical as well as in vitro and in vivo experimental studies on the mechanism by which the OX40/OX40L axis regulates the balance of T-cell subsets in OLP are still needed in the future.

Association of TNFSF4 polymorphisms with systemic lupus erythematosus: a meta-analysis

Abstract Objective: To more precisely estimate the association between the tumor necrosis factor ligand superfamily member 4 (TNFSF4) gene polymorphisms and systemic lupus erythematosus (SLE) susceptibility, we performed a metaanalysis on the association of the following single nucleotide polymorphisms (SNPs) of TNFSF4 with SLE: rs1234315, rs844648, rs2205960, rs704840, rs844644, rs10489265. Methods: A literature-based search was conducted using PubMed, MEDLINE, Embase, Web of Science databases, and Cochrane Library databases to identify all relevant studies. And the association of TNFSF4 gene polymorphisms and SLE susceptibility was evaluated by pooled odds ratio (OR) with 95% confidence interval (CI). Results: The meta-analysis produced overall OR of 1.42 (95% CI 1.36-1.49, P < 0.00001), 1.41 (95% CI 1.36-1.46, P < 0.00001) and 1.34 (95% CI 1.26-1.42, P < 0.00001) for the rs2205960, rs1234315 and rs704840 polymorphisms respectively, confirming these three SNPs confer a significant risk for the development of SLE. On the other hand, the meta-analysis produced overall OR of 0.92 (95% CI 0.70-1.21, P = 0.54) for the rs844644 polymorphism, suggesting no significant association. And no association was also found between either rs844648 1.11 (OR 1.11, 95% CI 0.86-1.43, P = 0.41) or rs10489265 (OR 1.17,95% CI 0.94-1.47, P = 0.17) polymorphism with SLE susceptibility, respectively. Conclusions: Our meta-analysis demonstrated that the TNFSF4 rs2205960, rs1234315 and rs844840 SNPs was significantly associated with an increased risk of SLE.

Establishment and validation of OX40/FcγR-humanized mice for the study of agonistic anti-OX40 antibody / 上海交通大学学报（医学版）

Objective • To establish a rapid method to evaluate the activity of agonistic antibody using OX40 (tumor necrosis factor receptor superfamily member 4)/FcγR (Fcγ receptor)-humanized mice. Methods • Bone marrow cells from OX40-humanized mice and FcγR-humanized mice were collected and mixed with equal ratio. Then the mixed bone marrow cells were administrated into irradiated wild-type mice through the tail veins. The reconstruction efficiency of the immune system was confirmed by detecting the expression of hOX40 and hFcγR in the immune cells of chimera mice. After the chimera mice were generated successfully, they were used to evaluate the immunostimulatory activity of anti-hOX40 antibodies to CD4+ or CD8+ T cells. The results of flow cytometry were statistically analyzed. The unpaired t-test was used to compare the means between the two groups, and oneway ANOVA was used to compare the means between multiple groups. Results • Flow cytometry analysis showed that wild-type recipient mice were efficiently reconstituted with hFcγR expressing cells and hOX40 expressing cells to generate OX40/FcγR-humanized bone marrow chimera mice. In these mice, B cells and myeloid cells expressed hFcγRs (P<0.05), and T cells expressed hOX40 upon in vitro stimulation (P<0.05). When these mice were used to evaluate the immunostimulatory activity of anti-hOX40 antibody, significant expressions of IFN-γ and hOX40 were observed (P<0.05). Conclusion • OX40/FcγR-humanized bone marrow chimera mice are generated based on hFcγR expressing cells and hOX40 expressing cells, suggesting a rapid method to build a mouse model with both hFcγR and hOX40 expression. These mice are suitable for evaluating the immunostimulatory activity of agonistic human anti-hOX40 antibodies.

Expression and significance of OX40/OX40L in peripheral blood of patients with autoimmune hepatitis, primary biliary cholangitis, and their overlap syndrome / 临床肝胆病杂志

ObjectiveTo investigate the expression and clinical significance of OX40/OX40L (CD134/CD134L) in CD4+ T cells, CD8+ T cells, monocytes, and B lymphocytes in peripheral blood of patients with autoimmune hepatitis (AIH), primary biliary cholangitis (PBC), and their overlap syndrome before and after standardized treatment. MethodsA total of 74 patients with AIH, PBC, and their overlap syndrome who were diagnosed in Subei People’s Hospital of Jiangsu from August 2015 to August 2019 were enrolled, and according to related diagnostic criteria, they were divided into AIH group (group A) with 29 patients, PBC group (group P) with 26 patients, and overlap syndrome group (group C) with 19 patients. A healthy control group with 30 individuals was also established. Peripheral blood samples were collected before and after standardized treatment to measure the expression of OX40/OX40L on the surface of peripheral blood cells by immunofluorescence flow cytometry, and the expression of OX40/OX40L was compared before and after treatment and between the three groups and the healthy control group to investigate its clinical significance. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups; the paired t-test was used for comparison of paired samples between two groups. ResultsThere were no significant differences in sex composition and age composition between the three groups (P＞0.05). Before treatment, the positive rate of OX40 in peripheral blood CD4+ T cells gradually increased in groups A, P, and C, and groups A, P, and C had a significantly higher positive rate of OX40 than the control group (14.80%±4.99%/17.11%±2.71%/25.18%±5.55% vs 6.67%±2.26%, F=14.823, P＜0.001); groups A, P, and C had a significantly higher positive rate of OX40 in CD8+ T cells than the control group (4.86%±1.54%/6.40%±1.88%/7.33%±2.12% vs 4.09%±2.69%, F=5.486, P＜0.001); the positive rate of OX40L in CD14+ monocytes was 19.84%±6.11% in group A, 21.17%±4.35% in group P, 29.13%±6.32% in group C, and 4.86%±2.34% in the control group, and there was a significant difference between groups (F=17004, P＜0.001); the positive rate of OX40L in CD19+ B cells was 17.62%±3.86% in group A, 14.75%±4.32% in group P, 1013%±2.56% in group C, and 4.50%±1.38% in the control group, showing a trend of gradual reduction, and groups A, P, and C had a significantly higher positive rate than the control group (F=12.221, P＜0.001). After treatment, the positive rate of OX40 in CD8+ T cells decreased significantly to a similar level as the control group, and there was no significant difference between groups (F=0731, P=0.538). For the other three types of cells, although there were varying degrees of reduction in the positive rate of OX40/OX40L after treatment, groups A, P, and C still had a significantly higher positive rate than the control group; in CD4+ T cells, the positive rate of OX40 was 11.00%±1.98% in group A, 13.72%±1.03% in group P, 19.72%±3.47% in group C, and 6.67%±2.26% in the control group, and groups A, P, and C had a significantly higher positive rate than the control group (F=11.365, P＜0.001); in CD14+ monocytes, the positive rate of OX40L was 11.82%±2.23% in group A, 15.19%±4.42% in group P, 24.51%±4.09% in group C, and 4.86%±2.34% in the control group, and groups A, P, and C had a significantly higher positive rate than the control group (F=13748, P＜0.001); in CD19+ B cells, the positive rate of OX40L was 9.09%±3.25% in group A, 6.81%±2.20% in group P, 748%±2.85% in group C, and 4.50%±1.38% in the control group, and groups A, P, and C had a significantly higher positive rate than the control group (F=8.052, P＜0.001). Groups A, P, and C had significant reductions in the expression of OX40/OX40L in peripheral blood CD4+ T cells, CD8+ T cells, CD14+ monocytes, and CD19+ B lymphocytes after treatment (all P＜0.05). ConclusionThe expression of OX40/OX40L in peripheral blood increases in patients with AIH, PBC, and their overlap syndrome and decreases after treatment, indicating that the OX40/OX40L pathway is involved in the pathogenesis of the above diseases, and the role of OX40 on the surface of CD8+ T cells may better reflect the treatment outcome.

Therapeutic strategies for the costimulatory molecule OX40 in T-cell-mediated immunity

The T cell co-stimulatory molecule OX40 and its cognate ligand OX40L have attracted broad research interest as a therapeutic target in T cell-mediated diseases. Accumulating preclinical evidence highlights the therapeutic efficacy of both agonist and blockade of the OX40-OX40L interaction. Despite this progress, many questions about the immuno-modulator roles of OX40 on T cell function remain unanswered. In this review we summarize the impact of the OX40-OX40L interaction on T cell subsets, including Th1, Th2, Th9, Th17, Th22, Treg, Tfh, and CD8 T cells, to gain a comprehensive understanding of anti-OX40 mAb-based therapies. The potential therapeutic application of the OX40-OX40L interaction in autoimmunity diseases and cancer immunotherapy are further discussed; OX40-OX40L blockade may ameliorate autoantigen-specific T cell responses and reduce immune activity in autoimmunity diseases. We also explore the rationale of targeting OX40-OX40L interactions in cancer immunotherapy. Ligation of OX40 with targeted agonist anti-OX40 mAbs conveys activating signals to T cells. When combined with other therapeutic treatments, such as anti-PD-1 or anti-CTLA-4 blockade, cytokines, chemotherapy, or radiotherapy, the anti-tumor activity of agonist anti-OX40 treatment will be further enhanced. These data collectively suggest great potential for OX40-mediated therapies.

Progress of OX40 and its ligand in lymphoma immunotherapy / 白血病·淋巴瘤

OX40 is a member of the tumor necrosis factor receptor (TNFR) superfamily. In the immune response of the body, OX40 and the OX40 ligand (OX40/OX40L) on the antigen-presenting cell membrane are important co-stimulatory molecules, which can promote the proliferation of T cells. And OX40 also has the dual role of activating and enhancing the T cell immune response. OX40/OX40L is an important target for tumor immunotherapy, and clinical studies of several OX40 agonists are currently underway. This article reviews the immunoregulatory mechanisms of OX40/OX40L and its research progress in lymphoma immunotherapy.

Research progress in OX40/OX40L-induced transplantation tolerance / 中华微生物学和免疫学杂志

OX40 (CD134) is a costimulatory molecule on the surface of T cells. It binds to OX40L (CD252) that is expressed on the surface of antigen presenting cells (APC) and participates in the transmission of the second signal of T cell activation, indicating that it is of great importance to the proliferation, differentiation, survival and migration of T cells. In theory, blocking the OX40/OX40L costimulatory pathway can suppress immune response and induce immune tolerance. In this review, we reviewed the research progress in OX40/OX40L-induced transplantation tolerance.

Relationship between Prognosis and Level of Plasma OX40L in Patients with Stable Angina Pectoris after Percutaneous Coronary Intervention / 中国医科大学学报

Objective To observe the ehanges in plasma OX40L level in patients with stable angina pectoris (SAP) after percutaneous coronary intervention (PCI) and to investigate the relationship between OX40L level and prognosis in patients with SAP.Methods We collected 200 cases of SAP after PCI.Plasma OX40L levels were examined at different time points,including before and after stent placement,in all patients withSAP.The patients with SAP were followed up for 18 months,and the end point was adverse cardiovascular events.Results The plasma OX40L level increased more apparently after PCI than before PCI (P ＜ 0.05) and peaked at 24 hours.We followed up 185 patients successfully,including 39 (21.08％) patients with and 146 (78.92％) without adverse cardiovascular events.The mean maximum OX40L level was higher in patients with than in those without adverse events (P ＜ 0.05).Multivariable logistic regression analysis revealed that the mean maximum OX40L level was related to adverse cardiovascular events (P ＜ 0.05).Conclusion PCI may cause upregulation of plasma OX40L expression.Mean maximum OX40L level showed good clinical predictive value of occurrence of cardiovascular events 18 months after PCI.

Advances in novel anti-OX40 cancer immunotherapy / 中华微生物学和免疫学杂志

OX40 (CD134) is a member of the tumor necrosis factor receptors and mainly expressed on the surface of active lymphocytes.Being an important pair of co-stimulatory factors in the process of immune response, OX40 and its ligand OX40L play an important role in inducing anti-tumor immune response.They regulate the immunosuppressive effect in tumor microenvironment and enhance the killing activities of effector cells.Pre-clinical animal studies have shown that agonists for OX40, whether they are used alone or in combination with other treatments, have stronger anti-tumor effects.Several anti-OX40 agonistic monoclonal antibodies (McAb) are currently tested in early phase cancer clinical trials.This review focuses on the advances in anti-OX40 mAbs-based therapy in terms of molecular and biological properties, mechanism of action, monotherapy and combined therapy.

Costimulate moleculars amplified the DRibble mediated T cells response / 中国药科大学学报

@#Autophagosomes derived from tumor cells have been proved to induce potent T cell response both in mouse and human. In human in vitro study, dendritic cells(DC)loaded with cytomegalovirus(CMV)pp65 antigen-containing DRibble were capable to efficiently re-stimulate pp65-specific T-cell recall responses from freshly isolated or frozen humanperipheral blood mononuclear cell(PBMC). This study developed more robust assays using in vitro expanded antigen-specific T cells that contained a much higher percentage of antigen-specific T cells. DC cross-presentation efficiency of OX40 and CD80 modified pp65-DRibble was detected by intracellular IFN-γ staining. Compared with Ctrl/pp65 DRibble, the percentage of IFN-γ+ in total CD8+ T cells andCD4+ T cells was improvedwith OX40/pp65 DRibbleand CD80/pp65 DRibble stimulation. In addition, vaccine induced IL-12indendritic cells, whichpolarizes Th cells toward the IFN-γ high Th1 phenotype, evaluated by ELISA inco-culture supernatantwas dramatically higher in OX40/pp65 DRibble and CD80/pp65 DRibblegroups than in Ctrl/pp65 DRibble group. These results have implications for the immuneactivity of OX40 and CD80 modified DRibble and choice for prospective clinical use ofDRibble-based cancer immunotherapy.

Ag85B regulates myeloid dendritic cell maturation and suppresses expres-sion of TSLPR and OX40L mediated by TSLP in vitro / 中国病理生理杂志

AIM:To investigate the maturation of mice immature myeloid dendritic cells (mDCs) induced by antigen(Ag)85B of mycobacterium tuberculosis, and the expression of TSLPR and OX40L mediated by TSLP in vitro. METHODS:Recombinant mouse GM-CSF ( rmGM-CSF) and rmIL-4 were used to induce bone marrow precursor cells of C57BL/6 mice to differentiate into immature mDCs in vitro.mDCs were identified followed by purification using CD 11c binding magnetic beads .The morphological characteristic of mDCs was observed under inverted phase-contrast microscope and scanning electron microscope .The surface phenotypes of mDCs were determined by flow cytometry .To obtain the opti-mal concentrations of Ag85B and TSLP, immature mDCs were cultured with different concentrations of Ag 85B or TSLP at 0 (control group), 50, 100 and 200 μg/L for 24 h, and the expression of cell surface molecules CD 80, CD86, TSLPR and OX40L was detected by flow cytometry.In addition, the expression of TSLPR and OX40L in Ag85B and TSLP-co-stimula-ted mDCs was determined by flow cytometry .RESULTS:After 7 d of culture in vitro, the cells showed irregular dendritic protrusions under the inverted-phase contrast microscope , and had wrinkles and dendritic splits under scanning electron mi-croscope , conformed to the morphological characteristics of immature mDCs .The mDCs cells expressed higher level of spe-cific marker CD11c, lower level of co-stimulatory molecules CD80 and CD86, which conformed to the phenotype of imma-ture mDCs.The CD80 +and CD86 +cell ratios of mDCs displayed significant increases in 50, 100 and 200μg/L Ag85B or TSLP groups compared with control group (P<0.05).The ratios of TSLPR +and OX40L+cells did not differ among dif-ferent concentrations of Ag 85B groups.The ratios of TSLPR +and OX40L+cells were significantly increased in 100 μg/L and 200μg/L TSLP groups compared with control group and 50μg/L TSLP group (P<0.05).Under the circumstance of optimal Ag85B or TSLP treatment concentration at 200 μg/L, there was significantly decreased in TSLPR and OX 40L cell ratio of mDCs in Ag85B group or Ag85B combined with TSLP group when compared with TSLP group (P<0.05), and no significant difference among Ag85B group, Ag85B combined with TSLP group and control group was observed .CONCLU-SION: Ag85B enhances mDCs maturation by up-regulating the expression of co-stimulatory molecules CD80 and CD86, and inhibit the expression of pro-inflammatory specific molecules TSLPR and OX40L on TSLP-activated mDCs, indicating that Ag85B modifies the development of asthmatic airway inflammation through the pathway of TSLP -activated mDCs.

Effects of Different Doses of Rosuvastatin on Serum Levels of sOX40L and MMP-9 in Patients with Acute Coronary Syndrome / 天津医药

Objective To explore effects of different doses of rosuvastatin on serum soluble OX40 ligand (sOX40L), matrix metalloproteinase-9 (MMP-9) in patients with acute coronary syndrome (ACS). Methods Sixty patients with ACS were evenly divided into 20 mg rosuvastatin treatment group (20 mg group) and 10 mg rosuvastatin treatment group (10 mg group). Twenty healthy volunteers were randomly assigned to control group. The serum levels of sOX40L and MMP-9 were measured and analyzed before and after treatment in three groups. Results The levels of sOX40L and MMP-9 before treat-ment were significantly higher in two treatment groups than those in control group (P<0.01). After two-week treatment with rosuvastatin, serum levels of sOX40L and MMP-9 decreased significantly in two treatment groups compared with those of pre-treatment (P<0.01). Moreover, the levels of sOX40L and MMP-9 were significantly lower in 20 mg group than those in 10 mg group after two-week treatment with rosuvastatin (P<0.05). But the levels of sOX40L and MMP-9 were still higher than control group (P<0.01). Conclusion Larger doses of rosuvastatin may decrease the degradation of extraceller matrix of coronary atherosclerotic plaque and inflammatory reaction, stabilize coronary atherosclerotic vulnerable plaque, and play an important role in improving prognosis in patients with ACS.

Ox-LDL increases OX40L in endothelial cells through a LOX-1-dependent mechanism

Oxidative low-density lipoprotein (Ox-LDL) is a key risk factor for the development of atherosclerosis, and it can stimulate the expression of a variety of inflammatory signals. As a new and highly sensitive inflammation index, OX40L may be a key to understanding the mechanisms that regulate interactions between cells within the vessel wall and inflammatory mediators during the development of atherosclerosis. To investigate whether Ox-LDL regulates OX40L expression through an oxidized LDL-1 receptor (LOX-1)-mediated mechanism, we investigated the effect of different concentrations of Ox-LDL (50, 100, 150 µg/mL) on endothelial cell proliferation and apoptosis. Stimulation with Ox-LDL increased OX40L protein 1.44-fold and mRNA 4.0-fold in endothelial cells, and these effects were inhibited by blocking LOX-1. These results indicate that LOX-1 plays an important role in the chronic inflammatory process in blood vessel walls. Inhibiting LOX-1 may reduce blood vessel inflammation and provide a therapeutic option to limit atherosclerosis progression.

Mice islet allograft tolerance induced by blockade of OX40/OX40L and CD40/CD154 costimulation pathway / 中华器官移植杂志

Objective To investigate the effects of blockade of OX40/OX40L costimulation pathway on mice islet allograft tolerance in CD40/CD154 costimulation pathway blockade mice.Methods C57BL/6 mice were induced into diabetes mellitus as recipients,and were transplanted with DBA/2 mice islets.The recipients were divided into four groups,(1) treated with IgG as controls,(2) anti-OX40L mAb,(3) anti-CD154,(4) combined treatment of anti-OX40L mAb and anti CD154mAb.The mean survival time (MST) of islet allograft was observed.The expression of OX40 in activated T cells of CD154 deficient mice was detected.Effector T cells were obtained from the spleen of CD154 deficient mice cultured with or without anti-OX40L mAb for 3 days.The proliferation of T cells was assayed.Results The MST in the control group,anti-OX40L mAb group,anti-CD154 mAb group and anti OX40L mAb + anti-CD154 mAb group was 19,22,48,and ＞150 days respectively (P ＜0.05).The OX40 expression was readily induced in the 66％ activated T effector cells.CD154 deficient T effector cells proliferation was inhibited by the addition of anti-OX40L mAb in the culture in a dose-dependent fashion.Conclusion The blockade of OX40/OX40L costimulation pathway can promote islet allograft tolerance in CD40/CD154 costimulation pathway blockade mice by inhibiting the proliferation of T cells.

Role of OX40 in mechanisms of memory T cells in islet transplant tolerance / 中华器官移植杂志

Objective To investigate the role of OX40 in the mechanisms of memory T cells in islet transplant tolerance.Methods The expression of OX40 on native, like memory and memory CD8+T cells was detected by RT-PCR. Splenic T cells from B6 mice were injected into Rag-/- mice via the tail vein, and the Rag-/- mice were divided into three groups (n=8 each): control group, given IgG; treatment group, given anti-OX40L; and OX40 knock-out group, given T cells from OX40 knock-out B6 mice spleen. All recipients were induced into diabetes mellitus model after adoptive transfer. Islet transplantation was performed on all Rag-/- mice as recipients. The mean survival time of islet was observed.Results The expression of OX40 in native T cells, like memory T cells and memory T cells was 2.87, 111.24 and 146.15 respectively. The expression of OX40 in like memory and memory T cells was higher than in native T cells (P<0.05). Comparison with control group , The mean survival time of the DBA/2 islet allografts in treatment group (130 days) and OX40 knock-out group (125 days) was significantly longer than in control group (21 days, P<0.05).Conclusion The OX40 expression is high in memory T cells. The mean survival time of the islet allografts can be prolonged by blocking OX40/OX40L pathway. OX40/OX40L pathway may be the key point of transplant tolerance.

Expression of OX40/OX40L mRNA in experimental allergic neuritis under influence of Rho-kinase inhibitor / 中华神经科杂志

Objective To study the expression of mRNA of OX40 and OX40L in the sciatic nerve,spleen,peripheral blood mononuclear cells and lymph nodes of EAN under the influence of Rho-kinase inhibitor.Methods All 54 female Lewis rats were divided into 3 groups:the EAN group,the EAN+ Rho-kinase inhibitor group and the complete Freund's adjuvant(CFA)group.The rats were sacrificed at 9,17 and 26 days after immunized.Ox40 and OX40L mRNA were detected by RT-PCR which came from spleens,sciatic nerves,peripheral blood mononuclear cells and lymphonodes.Results In EAN+ Rho-kinase inhibitor group,the mRNA expression of OX40 were 0.266±0.031,0.298±0.024 and 0.113±0.018 at 9.17 and 26 days in the sciatic nerve,the expression were 0.453±0.030,0.496±0.100 and 0.220±0.016 in the lymph nodes.The mRNA expression of OX40L were 0.247±0.018.0.298±0.026 and 0.165±0.013 in the sciatic nerve,the expression were 0.283±0.027,0.306±0.011 and 0.161±0.012 in the lymph nodes.The mRNA expression of OX40 and OX40L in EAN+Rho-kinase inhibitor group was lower than EAN group at the three time points(t=2.24-4.89,P＜0.05),and the demyelination and inflammation cells infiltrating were ameliorated in spinal nerve.CFA group didn't show any clinical manifestation.Conclusion Rho-kinase inhibitor may ameliorate tlIe development of EAN through inhabiting the OX40 and OX40L activation.

Heterogeneity of IL-22-producing Lymphoid Tissue Inducer-like Cells in Human and Mouse

Lymphoid tissue inducer (LTi) cells have been characterized in mouse as a key cell when secondary lymphoid tissues are organized during development and memory T cells are formed after birth. In addition to their involvement in adaptive immune responses, recent studies show that they contribute to innate immune responses by producing large amount of interleukin (IL)-22 against microbial attack. Here, we compare IL-22-producing LTi and LTi-like cells in human and mouse and discuss their heterogeneity in different tissues.

The role of OX40 in CD4+T cells cytokines production in ulcerative colitis / 中华内科杂志

Objective To investigate the expression of OX40 on CD4+T cells in patients with ulcerative colitis (UC)and the role of OX40/OX40L interaction for the cytokine production of lamina propria(LP)-CD4+T cells from UC.Methotis LP-CD4+T cells were purified.The expression of OX40 molecule was measured with FACS.LP-CD4+T cells were cultured with different stimuli and proliferation was assessed.The cytokines concentrations of the culture supernatant were detected.Results No difference of the OX40 expression was observed among the CD4+T cells from peripheral blood(PB)of UC patients,LP of non-inflammatory colonic tissue in UC patients and control PB.However.the expression of OX40 was significantly higher on LP-CD4+T cells from inflammatory colonic tissue in UC patients.In vitro culture with antigen presenting cells,the levels of IFNγ and TNFα secreted by LP-CD4+T cells from the inflammatory colonic tissue were significantly higher than those from the non-inflammatory colonic tissue(both P＜0.01).The levels of IFNγ and TNFα secreted by LP-CD4+T cells from the inflammatory colonic tissue were further increased by anti-OX40 MoAb stimulation.but suppressed significantly by adding anti-OX40L MoAb (compared with non stimulation,P＜0.01,respectively).The IFNγ and TNFα secretion of the LP-CD4+T cells from the non-inflammatory colonic tissue were not significantly different with and without anti-OX40 or anti-OX40L MoAbs stimulation.IL-4 and IL-10 produced by LP-CD4+T cells from the inflammatory or non-inflammatory colonic tissue were not significantly changed when adding different stimuli.Conclusions OX40 is highly expressed on LP-CD4+T cells from inflammatory colonic tissue in patients with UC.AntiOX40L MoAb can inhibit the proinflammatory cytokines secreted by these cells.It is indicated that OX40+T cells are involved in the immunopathological process in UC and blockage of the interaction of OX40 and OX40Lis a new strategy to be considered for the treatment of the disease.

Prolonged rat liver allograft survival by in vivo targeting OX40-siRNA OX40-OX40L co-stimulatory cascade blockade / 中华普通外科杂志

Objective To investigate the effect of blockading OX40-OX40L co-stimulatory signaling on the survival time of liver allograft in rat.Methods siRNA-expression vectors were constructed to targeting OX40.3～5 minutes before DA to Lewis orthotopic liver transplantation was performed,5×109 pfu of targeting OX40 siRNA plasmid DNA were diluted in 5 ml of phosphate buffered saline(PBS)and inlected intravenously into recipient Lewis rat over a period of 10 seconds.Serum IL-2 and IFN-γ levels were assayed by ELISA,and mix lymphocyte response(MLR)were tested by 3H-thymidine.Results The survival time of recipients in siRNA treatment group(74.0±9.3)was significantly longer than that in control group[(7.3±0.5)days].In experiment group,the inflammatory cell infihration and liver tissue structure destruction were very slight.The concentration of serum IL-2 was much lower in siRNA treatment group[(46±8.4)pg/ml]than that in control group[(286.5±14.6)pg/ml].Meanwhile,the concentration of serum IFN-γ was much lower in siRNA treatment group [(202.7±14.6)pg/ml]than that in control group[(1682.7±87.9)pg/ml].Conclusion Administration of OX40-siRNA can blockade OX40-OX40L co-stimulatory signaling pathway.hence inhibit the rejection of liver allograft.

Interference of OX40 gene expression induced by small interfering RNA / 中国免疫学杂志

Objective:To investigate the specific interference of OX40 gene expression induced by RNAi technique in 293T cell lines transfected with rat OX40 gene.Methods:293T cells were transfected with recombined plasmid pEGFP-N1-GFP/OX40,and the positive cell clones were selected by fluorescence protein observation and RT-PCR.One specific dicer siRNA targeted to OX40 mRNA was designed and synthesized,which shared no homology with exons of known human gene.Quantitative real-time PCR was performed to measure the inhibitory rate of target gene expression by comparing OX40 mRNA concentrations before and after siRNA transfection.Results:10 nmol/L siRNA-OX40 elicited the highest level of gene silence in 293T cells which was transfected with siRNA after 48 h (68.3?8.7)%);The time of maximal inhibitory effect was at 48-72 h [(61.7?8.4)%,(39.6?5.6)%].Conclusion:The exogenous OX40 expression can be significantly inhibited by treatment with specific siRNA in a dose and time -dependent manner in 293T cells,which may provide a useful profile for further investigation of inhibition of OX40 protein,and a promising control approach for preventing immune reaction.