足月胎盘单核-巨噬细胞体外诱导分化为树突状细胞

目的:研究人类足月胎盘中的单核-巨噬细胞是否能分化为树突状细胞(DCs).方法:机械分离足月胎盘中的单核-巨噬细胞,加入内皮细胞单层中,经正逆两次穿越内皮,48 h后收集返回到内皮单层上方得到的细胞,以扫描电子显微镜和普通光学显微镜观察细胞形态,流式细胞仪分析细胞表型及3H-TdR检测DCs激发异体T细胞增殖能力.结果:足月胎盘中的单核-巨噬细胞经过(逆)穿越内皮系统后,具有典型的DCs的形态学特征,相对于胎盘单核-巨噬细胞而言高表达CD80、CD86以及HLA-DR,并具有激发异体T细胞增殖能力.结论:足月胎盘中的单核巨噬细胞在内皮(逆)穿越系统中可以分化为DCs.     

EGCG 对Ro52 介导的THP-1 细胞凋亡 及炎症因子的影响

目的 探讨表没食子儿茶素没食子酸酯（EGCG）对SSA/Ro52（Ro52）介导的人单核- 巨噬 细胞株THP-1 凋亡及炎症因子分泌的影响。方法 构建稳定Ro52 过表达质粒（pCMV-Ro52）,pCMVRo52 转染THP-1 细胞48 h,检测Ro52 mRNA 和蛋白表达水平的变化,采用膜联蛋白- 荧光素异硫氰酸酯 和碘化丙锭双染检测THP-1 细胞凋亡率,酶联免疫吸附法检测肿瘤坏死因子-α（TNF-α）、白介素-1β （IL-1β）、IL-13 水平。pCMV-Ro52 质粒转染48 h 后, 分别使用0、5、10、20 和50 mg/L EGCG 处理细 胞24 h,检测Ro52 表达的变化、THP-1 细胞的凋亡,以及TNF-α、IL-1β、IL-13 水平。结果 pCMVRo52 促进Ro52 表达（P <0.05）,上调THP-1 细胞凋亡率及TNF-α、IL-1β、IL-13 水平（P <0.05）; 在Ro52 过表达的THP-1 细胞中,与0 mg/L EGCG 组相比,10、20 和50 mg/L EGCG 组Ro52 蛋白表达水 平、凋亡率及TNF-α、IL-1β、IL-13 水平降低（P <0.05）。结论 Ro52 过表达可促进人单核- 巨噬细胞 THP-1 凋亡及炎症因子TNF-α、IL-1β、IL-13 分泌;EGCG 可减弱Ro52 介导的THP-1 细胞凋亡,减 少炎症因子TNF-α、IL-1β、IL-13 的分泌。     

Cellular targets and receptors for interleukin-6 II. Characterization of IL-6 binding and receptors in peripheral blood cells and macrophages

Within 15 min, approximately 2.5% of 125I-labelled interleukin-6 (IL-6) injected intravenously into rats was taken up by the spleen. As determined by light microscopic autoradiography, uptake was mainly (60%) accounted for by macrophages in the red pulp. 125I-IL-6 binding in rat peritoneal macrophages was quantitatively similar to that in cultured human monocytes and T-cells. By comparison, IL-6 binding to polymorphonuclear granulocytes and freshly isolated monocytes was low. Stimulation with antigen, but not with mitogen (PWM), induced receptor presentation in B-cells, whereas antigen and mitogen downregulated the binding in T-cells. At 4 degrees C, labelled IL-6 bound to cells with a half-time of about 1.5 h. Binding appeared reversible, but dissociation was slow and incomplete. The apparent Kd for IL-6 binding was about 30 pmol l-1 in most cell types, however, values of approximately 120 pmol l-1 were obtained in polymorphonuclear granulocytes. At 37 degrees C, 125I-IL-6 was rapidly internalized by T-cells and monocyte-macrophages, and after a lag time, TCA-soluble radioactivity was released from the cells following a sigmoidal curve. Polyacrylamide gel electrophoresis of radiolabelled IL-6 cross-linked to its binding sites in T-cells, yielded receptor-ligand complexes with molar masses of 70-80 and 120-140 kg mol-1. This would agree with a dimeric conformation of the IL-6 receptor.     

Chronic sclerosing cholangitis and recurrent pulmonary infections in two brothers associated with cellular immunodeficiency and increased cytokine production

Primary sclerosing cholangitis has occasionally been associated with congenital immunodeficiency. Recently, sclerosing cholangitis has been found in the course of AIDS and cryptosporidium or cytomegalovirus have been observed chronically in the biliary tract, suggesting their role in the evolution to sclerosing cholangitis. We report on 2 brothers with immunodeficiency without any evidence of AIDS, who showed sclerosing cholangitis and recurrent pulmonary infections in the course of their disease. No bacterial microorganism could be cultured from the biliary juice. The serum of both patients contained measurable levels of TNFα. Supernatants from cultured peripheral blood mononuclear cells of the two brothers contained higher amounts of tumor necrosis factor (TNFα), interleukins 1α and 1β than did supernatants from cultures of normal individuals. The role of cytokines as an aggravating factor in immunodeficiency is discussed as well as their role in the inflammatory process.     

妊娠不同时期胎盘单核-巨噬细胞来源树突样细胞刺激脐血T淋巴细胞的特性比较

目的:观察妊娠不同阶段胎盘来源的树突样细胞(DC样细胞)刺激脐血淋巴细胞增殖活化特点的差异,进一步研究胎盘免疫细胞在妊娠耐受及分娩发动中的作用?方法:机械分离妊娠中?晚期胎盘中的单个核细胞,用磁珠分离法获取CD14+ 细胞,以内皮(逆)穿越系统诱导其分化为DC样细胞,并用流式细胞仪分析细胞表型,ELISA法检测细胞培养上清中白细胞介素(IL)-12?IL-10水平,CCK-8法检测其激发脐血淋巴细胞增殖能力,流式细胞术检测其所刺激的脐血T淋巴细胞内细胞因子?结果:足月妊娠胎盘来源的经诱导的细胞高表达与免疫激活有关的细胞表面标志物CD80?CD86;培养上清中检出较高水平的IL-12,极低水平的IL-10,具有较强的刺激脐血淋巴细胞增殖的能力;能刺激脐血T淋巴细胞分化形成较多的IFN-γ产生细胞,而分化形成的IL-10产生细胞则较少?但相比之下,中期妊娠胎盘来源的经诱导的细胞则低表达CD80?CD86(P < 0.05),培养上清中检出较高水平的IL-10(P < 0.05),但检测出的IL-12浓度极低(P < 0.05),刺激脐血淋巴细胞的能力较弱(P < 0.05);刺激脐血T淋巴细胞分化形成较多的IL-10产生细胞(P < 0.05),而分化形成的IFN-γ产生细胞较少(P < 0.05)?结论:妊娠不同时期胎盘来源的单核-巨噬细胞分化形成的DC样细胞免疫学特性不同,高表达CD80?CD86?IL-12的DC样细胞细胞在妊娠免疫耐受中止及分娩发动中发挥着一定的作用,而低表达CD80?CD86,高表达IL-10的细胞可能参与了妊娠过程中的免疫耐受?     

Cells infected with herpes Simplex Virus induce human monocyte-macrophages to produce interferon

Human monocyte-macrophages (M?) produced interferon when incubated with human fibroblast, Vero cell, or rabbit kidney cell cultures infected with herpes simplex virus (HSV). Induction of M? interferon required a 12-hour incubation with the HSV-infected cultures. The interferon produced biologically resembled human leukocyte-derived rather than human fibroblast-derived interferon.     

妊娠不同时期胎盘单核-巨噬细胞免疫学特性的比较

目的:通过比较妊娠不同时期胎盘树突状细胞(dendritic cells,DCs)的前体细胞——单核-巨噬细胞的免疫学特性的差异,探索胎盘免疫细胞及微环境在妊娠耐受和分娩发动中的可能作用?方法:机械法分离中?晚期胎盘中的蜕膜组织细胞,密度梯度离心法提取外周血单个核细胞(PBMC),经CD14+免疫磁珠分选,获得胎盘组织的单核-巨噬细胞?流式细胞仪分析其细胞表型,ELISA检测其细胞培养上清中的细胞因子?再将分离获得的胎盘组织单核-巨噬细胞与同种异体淋巴细胞混合培养,以CCK-8法检测单核-巨噬细胞激发同种异体淋巴细胞增殖的能力,ELISA检测混合培养上清中的细胞因子?结果:自胎盘蜕膜组织中分离获得的单核-巨噬细胞纯度达到93%左右?足月胎盘来源的单核-巨噬细胞低表达与细胞免疫激活有关的细胞表面标志物CD80?CD86? HLA-DR,不表达CD83;细胞培养上清中低表达IL-10?TGF-β;具有较弱的刺激同种异体淋巴细胞增殖的能力,与同种异体淋巴细胞混合培养,上清中低表达IL-10,高表达TGF-β?相比之下,中期妊娠胎盘单核-巨噬细胞CD80?CD86的表达水平较高,以HLA-DR更显著(P < 0.05),CD83也有微弱的表达;细胞培养上清中高表达IL-10?TGF-β(P < 0.05);其刺激同种异体淋巴细胞增殖的能力也很弱,混合培养上清中高表达IL-10?TGF-β?结论:作为DCs的前体细胞,妊娠不同时期的单核-巨噬细胞在不同的子宫胎盘环境中已处于不同的状态?中期妊娠胎盘来源的单核-巨噬细胞可能通过IL-10发挥免疫抑制功能?     

Accelerated development of human monocyte-macrophages cultured on Plastek-C tissue culture dishes

Summary Many investigators have found that the surface on which cells are cultured can be an important factor in their adherence and subsequent growth. To enhance the development of monocyte-macrophage cultures, we have grown these cells on polystyrene tissue cultureware that has been treated with gas plasma to induce a positively charged culture surface (Plastek-C cultureware from Tekmat). Human monocyte-macrophages grown on Plastek-C tissue cultureware demonstrated a greater cell protein content during culture development as compared to monocyte-macrophages grown on standard polystyrene tissue cultureware. Part of the reason for enhanced development of human monocyte-macrophage cultures on Plastek-C cultureware seemed to be due to a higher density of cells maintained during the initial stages of culture development.     

APOPTOSIS IN HUMAN MONOCYTE-MACROPHAGES EXPOSED TO OXIDIZED LOW DENSITY LIPOPROTEIN

This study has demonstrated the toxicity to human monocyte-macrophages of low-density lipoprotein (LDL) which had been artificially oxidized using copper sulphate. The assays of cell damage used were tritiated adenine release, neutral red staining, lactate dehydrogenase leakage, and MTT dye reduction. Toxicity was concentration- and time-dependent. Exposure to native LDL under the same conditions did not result in toxicity. Transmission electron microscopy of cells exposed to oxidized LDL showed characteristic changes of apoptosis, including chromatin condensation and a decrease in cell volume. There was extensive loss of cell surface protrusions and evidence of the phagocytosis of apoptotic cells by neighbouring monocyte-macrophages. Apoptotic features preceded the increased membrane permeability revealed by the release of radioactivity from cells preloaded with tritiated adenine and by lactate dehydrogenase leakage. DNA fragmentation was indicated by nick end-labelling using the terminal transferase enzyme (TUNEL). The number of TUNEL-positive cells was markedly greater in cells exposed to oxidized LDL, compared with those incubated as no-additions controls. Inhibition of de novo protein synthesis with cycloheximide and of Ca²⁺/Mg²⁺-activated endonuclease activity with aurintricarboxylic acid or zinc ion did not inhibit the toxicity produced by oxidized LDL.