Toll-like Receptors and Lupus Nephritis

A variety of immune mechanisms, both humoral and cellular, are involved in the onset and amplification of the inflammatory response in lupus nephritis (LN). Accumulating evidence substantiates the view that innate immunity pathways may also amplify inflammatory reactions within the kidneys. Toll-like receptors (TLRs) are essential modulators of the innate immune response thanks to their ability to rocognize conserved molecular patterns that are microbe specific and other danger signals. Their recognition of endogenous molecules released from injured cells may also contribute to renal inflammation. Studies conducted in vivo and in vitro provide experimental evidence for the functional role of TLRs in LN. Intriguingly, these data suggest that pharmacological TLR signal suppression could be a useful approach to the treatment of systemic lupus erythematosus.     

Toll-like Receptors in Pregnancy Disorders and Placental Dysfunction

The Toll receptor was originally identified as a regulator of embryogenesis in Drosophila. Toll-like receptors (TLRs) in mammals recognize infectious agents and other danger signals. Activation of TLRs on trophoblast influences immune cell recruitment, cytokine secretion, and decidual responses to invading pathogens during pregnancy. Importantly, biological effects of TLR signal transduction at multiple maternal–fetal interfaces may contribute to several pregnancy pathologies associated with placental dysfunction, including pre-eclampsia, intrauterine growth restriction, and preterm labor. We herein discuss mechanisms by which TLRs regulate the maternal immune response during normal and abnormal gestation, and we highlight recent data that assign a role to TLRs in the pathophysiology of selected pregnancy-associated complications.     

Toll-like Receptors and RBL-2H3 Mast Cells

Inflammation Research -     

Toll样受体与Th1和Th2型免疫应答

Toll样受体是广泛表达在哺乳动物细胞表面的跨膜信号传导受体,其通过识别多种类型的病原体相关分子模式及一些内源性配体,激活天然免疫系统,同时通过诱导树突状细胞分化成熟,调控获得性免疫反应的建立。大多数TLRs的配体可诱导机体产生Th1型免疫应答,然而在某些条件下也可导致Th2型免疫应答的发生。弄清TLRs参与调节Th0分化的机制,可为今后在感染免疫、自身免疫、超敏反应等方面进行深入研究及相关疾病的治疗提供新的切入点。     

Elucidation of the Receptors in Macrophage and Plasmodium Yoelii Interactions.

Binding of hyperimmune serum opsonized merozoites of Plasmodium Yoelii nigerensis to trypsinized macrophages suggested it to be mediated by FcII receptor. Receptor blocking inhibition with monoclonal antibody 2.4G2 directed against Fc receptor for IgG1/IgG2b provided evidence that Fcγ2b on macrophage played an important role in the merozoite-macrophage interactions. In addition, a neuraminidase sensitive receptor was noted to mediate the binding of P. yeelii merozoites in the absence of serum. Binding inhibition studies with two monosaccharides, D-mannose and α-methyl mannoside, indicated the role of Mannose/Fucose receptor on macrophage in this interaction.     

The Fc Receptors of Normal and Cancer Patients Monocytes

The ability of human monocytes from normal donors and gastric-cancer patients to form rosettes with ?0? Rh⁺(D) human erythrocytes coated with hyperimmune IgG anti-D antibody (EA_hu) and to kill the same target in antibody-dependent cellular cytotoxicity (ADCC) were assessed. Trypsin pretreatment of normal monocytes decreased their ability to form rosettes with EA_hu complexes, but their ADCC activity was unaffected. The Fc receptor (FcR) expression and ADCC activity of monocytes of cancer patients were elevated, and trypsin-treatment led to their further increase. The elevated values were related to the presence of the tumour. These results may suggest that human monocytes possess trypsin-sensitive and trypsin-resistant Fc receptors. The trypsin-resistant FcR seems to be involved in ADCC phenomenon and to be preferentially expressed on monocytes of some cancer patients.     

幽门螺杆菌VacA通过激活NF-kB诱导巨噬细胞分泌及凋亡

目的 研究幽门螺杆菌(Helicobacterpylori)空泡毒素(VacA)单一毒力决定簇对THP-1巨噬细胞分泌和凋亡的影响,以及核因子KB(nuclear factor KB,NF-KB)在调节VacA诱导的THP-1巨噬细胞功能中的作用.方法 将pDsRed-Monomer-Cl/vacA转染THP-1巨噬细胞,ELISA法检测巨噬细胞培养上清IL-1β、TNF-α含量;Griess试剂分析培养上清的一氧化氮(NO)水平;荧光探针DCFH-DA检测培养上清的活性氧(ROS);流式细胞仪检测细胞的凋亡率;凝胶阻滞试验(electro-phoretic mobility gel shift assay,EMSA)检测NF-kB的核转位.结果 重组质粒转染后6 h,重组质粒组培养上清中TNF-α、IL-1β含量明显高于阴性对照组(P<0.05);且分别于转染后6 h或24 h到达峰值;转染后6 h或12 h,重组质粒组培养上清中NO、ROS含量明显高于阴性对照组(P<0.05),且于转染后24 h达到高峰.转染后16 h,重组质粒组的凋亡率明显升高,与阴性对照组相比,差异有统计学意义(P<0.05).重组质粒组加NF-kB的特异性抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)后,IL-1β、TNF-α、NO、ROS的分泌量和细胞的凋亡率明显降低,与重组质粒组相比差异有统计学意义(P<0.05).EMSA试验显示,转染后3h细胞核内有活化的NF-kB,且于转染后12h活性最强.结论 VacA蛋白瞬时高表达上调THP-1巨噬细胞分泌IL-1β、TNF-α、NO和ROS;VacA蛋白瞬时高表达诱导THP-1巨噬细胞凋亡;NF-kB可能参与调节VacA诱导的THP-1巨噬细胞的分泌和凋亡.     

抑制核因子-κB活性对TNF-α诱导的肾小球系膜细胞血管紧张素原表达的影响

目的： 研究抑制核因子-κB（NF-κB）活性对肿瘤坏死因子-α（TNF-α）诱导的SD大鼠肾小球系膜细胞血管紧张素原表达及血管紧张素Ⅱ产生的影响。方法：分离SD大鼠肾小球系膜细胞分为下列3组：对照组,TNF-α处理组,TNF-α＋NF-κB抑制剂吡咯烷二硫氨基甲酸酯（PDTC）处理组（TNF-α＋PDTC处理组）。以电泳迁移率变动分析法（electrophoretic mobility shift assay,EMSA）检测NF-κB 活性,放射免疫分析法检测培养液血管紧张素Ⅱ水平,RT-PCR方法检测血管紧张素原mRNA表达,蛋白质印迹法（Western blotting）检测血管紧张素原蛋白表达。结果： TNF-α处理组NF-κB活性［（20.67±9.14）×102 μg/cell］显著高于对照组［（8.25±4.35）×102 μg/cell,P<0.01］与TNF-α＋PDTC处理组［（7.20±4.57）×102 μg/cell,P<0.01］,TNF-α＋PDTC处理组与对照组比较无显著差异。血管紧张素原mRNA表达TNF-α处理组（0.27±0.05）显著高于对照组（0.20±0.05,P<0.05）,与TNF-α＋PDTC处理组（0.22±0.06,P>0.05）比较无显著差异;蛋白表达TNF-α处理组［（0.60±0.19） μg/cell］显著高于对照组［（0.37±0.15） μg/cell,P<0.05］及TNF-α＋PDTC处理组［（0.37±0.17） μg/cell,P<0.05］,TNF-α＋PDTC处理组与对照组比较无显著差异（P＞0.05）。培养液血管紧张素Ⅱ水平在TNF-α处理组［（9.73±2.38）×10-5 ng·L-1/cell］显著高于对照组［（7.50±1.51）×10-5ng·L-1/cell,P<0.05］及TNF-α＋PDTC处理组［（6.94±1.46）×10-5 ng·L-1/cell,P<0.05］,TNF-α＋PDTC处理组与对照组比较无显著差异（P＞0.05）。结论：TNF-α可激活SD大鼠肾小球系膜细胞NF-κB,并诱导血管紧张素原表达及血管紧张素Ⅱ产生增加;抑制NF-κB活性可降低血管紧张素原表达及血管紧张素Ⅱ产生。结果提示NF-κB介导TNF-α诱导的肾小球系膜细胞血管紧张素原表达及血管紧张素Ⅱ产生。     

Chimeric Antigen Receptors in Cancer Immuno-Gene Therapy: Current Status and Future Directions

The concept of chimeric antigen receptors (CARs) as molecules able to redirect T lymphocytes toward tumor cells is currently being exploited in the field of cancer immunotherapy. Despite promising preliminary results, some clinical trials evidenced limitations for this technology that must be overcome for more extensive application of CARs in tumor immunotherapy. We describe here the fundaments of these molecules in terms of structure, function, possible targets and pre-clinical and clinical applications. We also discuss strategies that can potentially overcome the limitations seen so far, paving the road to a wider application of this exciting new technology.     

The Expression of Toll-like Receptor 8 and Its Relationship with VEGF and Bcl-2 in Cervical Cancer

BACKGROUND: Cervical cancer is one of the most common cancers in women worldwide, often associated with the infection of human papillomavirus (HPV). Toll-like receptor 8 (TLR8), a pattern recognition receptor, is involved in viral nucleic acid sensing. Recently TLR8 has been shown to be expressed in cancer cells, and it has been suggested that it may help cancer cell growth and tumor development. The objective of this study is to investigate the expression of TLR8 expression and its relationship with Bcl-2 and VEGF in cervical cancer cells.METHODOLOGY/PRINCIPAL: The mRNA expression levels of Bcl-2, VEGF and TLR-7,-8,-9 in newly diagnosed cervical cancer patients were detected by quantitative real-time PCR (qRT- PCR). Epifluorescence microscope was used to determine the presence of TLR8 protein in Hela cells. The cell cycle and apoptosis were analyzed by flow cytometer, and the cell proliferation was measured by MTT assay. Our data showed the increased mRNA levels of TLR8 in human cervical cancer samples as well as in HeLa cells, a cell line derived from a human cervical cancer. In addition, there was a positive correlation between the expression levels of TLR8 and Bcl-2 and VEGF in cervical cancer patients. When Hela cells were treated with TLR8 agonist CL075, the percentage of cells in G2/M +S was remarkably increased, accompanied by increased COX-2, BCL-2 and VEGF mRNA levels.CONCLUSIONS/SIGNIFICANCE: The mRNA expression level of TLR8 in the patients with cervical cancer and Hela cells were up-regulated, it consistent with the increased expression of VEGF and Bcl-2. The results suggest that TLR8 may be an interesting therapeutic target in cervical cancer.     

TRAIL及NF-kB在骨肉瘤中的表达及其意义

本文探讨TRAIL及NFkB在骨肉瘤中的表达及其与细胞增殖调控的关系。对16例非化疗骨肉瘤,5例骨巨细胞瘤和6例软骨肉瘤石蜡切片,应用流式细胞术(FCM)定量检测TRAIL及NFkB的表达。结果表明TRAIL和NFkB在骨肉瘤各期间的表达显著高于骨巨细胞瘤和软骨肉瘤(P<0.01);TRAIL和NFkB在骨肉瘤不同分化组间表达无明显差异(P>0.05);TRAIL和NFkB在骨巨细胞瘤和软骨肉瘤间表达无明显差异(P>0.05)。结论是TRAIL在骨肉瘤中高表达,但可能受NFkB高表达的调控,不能诱导细胞凋亡;在骨肉瘤的细胞增殖调控中,NFkB可能通过调控TRAIL诱导的凋亡途径进而抑制细胞凋亡的发生。     

Toll样受体与造血

Toll样受体（TLR）是近年来发现的一类天然免疫受体,在天然免疫及获得性免疫过程中均发挥非常重要的作用。对TLR的深入研究表明,TLRs在造血相关细胞（造血干／祖细胞、间充质干细胞、血管内皮细胞等）上均有不同程度的表达,并且发挥一定生物学效应。