人IL—1,IL—6及TNFα基因在卵巢癌患者单个核细胞中的表达与相互关系

利用ＭＴＴ法及原位杂交技术，检测了４６例卵巢癌患者外周血单个核细胞（ＰＢＭＣ）及腹水细胞分泌的ＩＬ－１、ＩＬ－６和ＴＮＦα蛋白水平及其ｍＲＮＡ的表达。结果表明，卵巢癌患者血ＰＢＭＣ细胞、腹水细胞的ＩＬ－１、ＩＬ－６和ＴＮＦα分泌水平明显高于健康对照组，具有统计学意义。且加入一定剂量的ＩＬ－１能促进这些细胞的ＩＬ－６及ＴＮＦα分泌水平，这种作用能被ＩＬ－１的单克隆抗体封闭。提示：ＩＬ－１、ＩＬ－６和     

透析营养不良的生化及肌测量改变及其与单核细胞产生白介素-1β及肿瘤坏死因子-α的关系探讨

探讨血液透析相关性营养不良与单核细胞产生的白细胞介素－１β（ＩＬ－１β）及肿瘤坏死因子α（ＴＮＦ－α）的关系。方法：对１５倒尿毒症血液透析患者、１２倒尿毒症非透析患者及２３例正常对照组进行研究,测定各组受检者上臂肌围（ＡＭＣ）、血清白蛋白、透析前后ＢＵＮ及尿尿素氮,计算校正蛋白分解率（ＮＰＣＲ）、时间平均尿素氮浓度（ＴＡＣｕｒｅａ）及ＫＴ／Ｖ值,同时测定上述３组患者单核细胞产生的白细胞介素－１β（ＩＬ－１β）及肿瘤坏死因子α（ＴＮＦ－α）的生物活性。结果与结论：透析营养不良患者的单核细胞的ＩＬ－１β及ＴＮＦ－α的生物活性升高,较其它两组有显著性差异（Ｐ＜０．０１）。     

氨甲喋呤对类风湿性关节炎患者外周血单个核细胞产生细胞因子的影响

目的研究氨甲喋呤（ＭＴＸ）对类风湿性关节炎（ＲＡ）患者外周血单个核细胞（ＰＢＭＣ）产生细胞因子的影响。方法采用ＥＬＩＳＡ双抗夹心法，观察ＲＡ患者ＴＮＦ－α、ＩＬ－６的自发分泌及ＭＴＸ和ＬＰＳ的影响，以及ＭＴＸ和ＰＨＡ对ＩＬ－１０和ＩＦＮ－γ产生的影响。结果低浓度ＭＴＸ（５ｍｇ·Ｌ－１）有抑制ＲＡ患者ＰＢＭＣ自发分泌ＩＬ－６的作用，并对ＬＰＳ（１０ｍｇ·Ｌ－１）诱导ＩＬ－６的产生具有抑制作用，对ＴＮＦ－α的自发分泌及ＬＰＳ促分泌作用无明显影响；而高浓度ＭＴＸ（１５ｍｇ·Ｌ－１）对ＴＮＦ－α、ＩＬ－６和ＩＮＦ－γ均具有抑制作用；并能促进ＰＨＡ（１０ｍｇ·Ｌ－１）诱导ＩＬ－１０的产生；使ＩＬ－１０／ＩＮＦ－γ的比率上升。结论ＭＴＸ通过调节细胞因子网络（增高Ｔｈ２型细胞产生的细胞因子和降低Ｔｈ１型细胞产生的细胞因子）来发挥免疫调节作用和抑制炎症反应，这可能是其对ＲＡ产生治疗作用机制之一     

白介素2和肿瘤坏死因子α对大鼠C6胶质瘤细胞产生一氧化氮的影响

用Ｇｒｉｅｓｓ法检测细胞培养上清中ＮＯ÷２含量作为反映细胞产生一氧化氮（ＮＯ）量的指标，观察细菌脂多糖（ＬＰＳ），干扰素γ（ＩＦＮ－γ），肿瘤坏死因子α（ＴＮＦ－α）及白介素２（ＩＬ－２）对大鼠Ｃ６胶质瘤细胞产生ＮＯ的影响．结果Ｃ６细胞于体外培养８ｈ时可自发产生ＮＯ，２４ｈ达峰值（１７．５±１．９ｖｓ溶剂对照组６．５±１．９μｍｏｌ·Ｌ－１），４８ｈ仍有产生．在所用剂量范围，上述因素单独作用２４ｈ均不影响Ｃ６产生ＮＯ，而５－５００ｋＵ·Ｌ－１ＩＬ－２与０．５ｍｇ·Ｌ－１ＬＰＳ或５０ｋＵ·Ｌ－１ＩＦＮ－γ合用可增强Ｃ６产生ＮＯ（１０．６－１３．４ｖｓ７．２μｍｏｌ·Ｌ－１），ＬＰＳ，ＩＦＮ－γ及ＴＮＦ－α三者合用亦有一定促进Ｃ６产生ＮＯ作用．提示炎性刺激与炎性细胞因子共同作用可促进中枢神经胶质细胞产生ＮＯ．     

缺氧／再给氧和细胞因子对白细胞与脑微血管内皮细胞粘附作用？…

目的：研究缺氧／再给氧（Ｈ／Ｒ）对中性粒细胞（Ｎｅｕ）和单核细胞（Ｍｏｎ）与诱导的培养牛脑微血管细胞粘附作用的影响。方法：牛脑微血管内皮细胞（ＣＭＦＣ）和平滑肌细胞（ＣＭＳＭＣ）经Ｈ／Ｒ处理或经ＴＮＦ－α和ＩＬ－１α刺激，Ｍｏｎ和Ｎｅｕ与ＣＭＥＣ和ＣＭＳＭＣ粘附的细胞数目用流式细胞仪测定，结果：ＣＭＥＣ先缺氧２ｈ再经复氧与ＴＮＦ－α（２μｇ．Ｌ＾－１）作用２ｈ，Ｍｏｎ和Ｎｅｕ与ＣＭＥＣ的粘附率分别     

一氧化氮和白细胞介素—10对小鼠肺泡巨噬细胞产生肿瘤坏死因？…

目的 观察一氧化氮和ＩＬ－１０对肺泡巨噬细胞炎症反应的调节作用,方法：小鼠肺泡汇噬细胞（ＡＭ）受脂多糖（ＬＰＳ）１０ｍｇ．Ｌ＾－１刺激同时,加入一氧化氮合酶抑制剂Ｓ－硫酸甲基异硫脲（ＳＭＴ）或一氧化氮供体Ｓ－亚硝基乙酰青霉胺（ＳＮＡＰ）,ＥＬＩＳＡ法测定上清液中ＴＮＦα,ＩＬ－１β,ＩＬ－６和ＩＬ－１０浓度,结果：ＡＭ受ＬＰＳ刺激后,ＴＮＦα,ＩＬ－１β和ＩＬ－６释放峰值分别在６,１２和２４小时,     

Cytokine and nitric oxide production by rat microglia stimulated with lipopolysaccharides in vitro

目的：研究ＬＰＳ刺激体外培养的新生大鼠小胶质细胞产生ＩＬ１，ＩＬ２，ＴＮＦα和ＮＯ的特征．方法：小胶质细胞与ＬＰＳ（０－１０ｍｇ·Ｌ－１）孵育０－７２ｈ，分别测定细胞外和细胞内的ＩＬ１，ＩＬ２和ＴＮＦα的生物活性和细胞外ＮＯ水平．结果：ＩＬ１，ＴＮＦα和ＮＯ分别在ＬＰＳ刺激后１，４，和８ｈ检测到，并在８，２４和２４ｈ达到峰值．ＬＰＳ１ｍｇ·Ｌ－１刺激细胞外ＩＬ１，ＴＮＦα和ＮＯ的产生最高，但细胞内ＴＮＦα水平极低，ＬＰＳ未能刺激ＩＬ２产生．结论：体外ＬＰＳ刺激大鼠小胶质细胞产生大量炎性细胞肽和ＮＯ．     

1α,25-二羟维生素D_3对患者PBMC分泌的IFN-α和IL-10水平的影响

目 的 探 讨 １α，２５－二羟 维生 素 Ｄ_３（１α，２５－（ＯＨ ）２Ｄ ）对 系 统性 红斑 狼 疮（ＳＬＥ）患 者 外周 血 单个 核 ３细 胞（ＰＢＭ Ｃ）分 泌的 白细 胞 介素 （ＩＬ）－１０ 和 干扰 素 （ＩＦＮ ）－α水平 的 影响 。方法 ２０ 例 ＳＬＥ 患 者和 １０ 名健 康 志愿者 ＰＢＭ Ｃ 的提 取 采 用 Ｆｉｃｏｌ密 度 梯 度 离 心 法 ，ＳＬＥ 组 和 对 照 组 在 不 加 或 加 入 １α，２５－（Ｏ Ｈ ）２Ｄ３ 的 情 况 下 孵 育 ７２ ｈ收 集培 养 上清 ，上 清液 ＩＬ－１０ 和 ＩＦＮ－α水平 检 测采 用酶 联 免疫 吸附 试 验（ＥＬＩＳＡ ）。结 果 与 正常 对 照组 相比 ，ＳＬＥ组 ＰＢＭ Ｃ 中 ＩＬ－１０ 和 ＩＦＮ－α水 平 明 显 增高 （Ｐ＜０．０１）。０．０１ ｍ ｏｌ／Ｌ １α，２５－（Ｏ Ｈ ）２Ｄ３ 的 加 入 明 显 抑 制 了 ＳＬＥ 组ＰＢＭ Ｃ 中 ＩＦＮ －α的 产 生 （Ｐ＜０．０１），增 加 了 ＳＬＥ 组 ＰＢＭ Ｃ 中 ＩＬ－１０ 的 产 生 （Ｐ＜０．０１），但 对 正 常 对 照 组 ＩＦＮ －α和ＩＬ－１０ 水平 无明 显 影响 。结 论 １α，２５－（ＯＨ ）２Ｄ 可 能 抑制 体外 培 养的 ＳＬＥ 患 者 ＰＢＭ Ｃ ＩＦＮ －α的产 生     

甲硫氨酸脑啡肽增强TNF—α产生,NK细胞活性和IL—12p35mRNA的表达

研究甲硫氨酸脑啡对机体免疫监督功能的影响。测定ｍｅｔ－ｅｎｋ对ＮＫ活性，抗癌细胞因子如：ＴＮＦ－α和ＩＬ－１２的产生和基因表达的影响。Ｍｅｔ－ｅｎｋ（１＾１０＾－８－１×１０＾－５ｍｏｌ．Ｌ＾－１）能增强ＮＫ细胞活性。体外，体内均能刺激ＴＮＦ－α的产生。     

Inhibitory effects of nitric oxide and interleukin-10 on production of tumor necrosis factorα,interleukin-1β,and interleukin-6 in mouse alveolar macrophages

目的：观察一氧化氮和ＩＬ１０对肺泡巨噬细胞炎症反应的调节作用．方法：小鼠肺泡巨噬细胞（ＡＭ）受脂多糖（ＬＰＳ）１０ｍｇ·Ｌ－１刺激同时,加入一氧化氮合酶抑制剂Ｓ硫酸甲基异硫脲（ＳＭＴ）或一氧化氮供体Ｓ亚硝基乙酰青霉胺（ＳＮＡＰ）．ＥＬＩＳＡ法测定上清液中ＴＮＦα、ＩＬ１β、ＩＬ６和ＩＬ１０浓度．结果：ＡＭ受ＬＰＳ刺激后,ＴＮＦα、ＩＬ１β和ＩＬ６释放峰值分别在６、１２和２４小时．ＳＭＴ抑制一氧化氮释放,但促进ＩＬ１β和ＩＬ６释放,对ＴＮＦα无影响．ＳＮＡＰ对ＩＬ１β和ＩＬ６释放有明显的抑制作用,呈剂量依赖效应．重组ＩＬ１０抑制ＴＮＦα、ＩＬ１β和ＩＬ６释放,而ＩＬ１０单克隆抗体促进上述因子释放．结论：内源及外源性一氧化氮和ＩＬ１０均对ＬＰＳ诱导的炎症性细胞因子释放有抑制作用．     

Pentoxifylline inhibits ICAM-1 expression and chemokine production induced by proinflammatory cytokines in human pulmonary epithelial cells

Airway epithelium participates in inflammatory reactions by producing chemokines and expressing cell-surface adhesion molecules which aid in the selective recruitment of effector cells. Previous studies showed that proinflammatory cytokines, interleukin 1 (IL-1) and tumor necrosis factor alpha (TNF alpha), induce surface expression of intercellular adhesion molecule 1 (ICAM-1) and the production of the chemokines interleukin 8 (IL-8) and monocyte chemoattractant protein (MCP-1) on pulmonary epithelial cell lines in vitro. In this study, the dose response of four cytokines, IL-1alpha, IL-1beta, TNF alpha and TNF beta, in inducing ICAM-1 expression and production of IL-8 and MCP-1 on pulmonary A549 epithelial cells was examined. Both IL-1alpha and IL-1beta induced ICAM-1 expression and IL-8 and MCP-1 production at lower doses than TNF alpha or TNF beta. Pentoxifylline, an anti-inflammatory agent used to treat vascular diseases, was tested for its ability to inhibit the activation of airway epithelial cells by these cytokines. Pentoxifylline completely inhibited the surface expression of ICAM-1 and the production of IL-8 and MCP-1 by cytokine-activated epithelial cells. As elevated levels of chemokines are often present in bronchial lavage fluids of patients suffering from various acute respiratory diseases, pentoxifylline may be useful for preventing the rapid development of immune reactions leading to lung injury.     

17β-雌二醇对人的类成骨细胞株TE85和U2功能的调节

目的：以人的类成骨细胞株TE85和U2为细胞模型，观察雌激素对细胞功能的影响。方法：³H-胸腺嘧啶参入法测细胞增殖；ELISA法测细胞因子IL-6含量；精氨酸转化法测iNOS活性。结果：IL-1(20 U.mL^-1)和TNFα(50 U.mL^-1)刺激细胞分泌IL-6，并抑制细胞增殖。在TE85细胞，17β-雌二醇(E₂)可抑制IL-6分泌并能对抗IL-1和TNFα对细胞增殖的影响。NOS抑制剂L-NMMA使细胞³H-胸腺嘧啶的参入降低，E₂(1 nmol.L^-1)可对抗L-NMMA的抑制作用。结论：E₂通过减少IL-6分泌和影响iNOS活性而调节细胞功能。     

铁杉灵芝多糖FⅢ-2对人组织瘤细胞和人外周血白细胞产生炎症性细胞因子的影响(英文)

用放射免疫分析法及逆转录聚合酶链反应法 ,比较研究水不溶性铁杉灵芝多糖 (FⅢ 2 )及其吡啶 氯磺酸修饰产物 (FⅢ 2 S)对人炎症性细胞因子蛋白质及其mRNA产生的影响 .结果表明 ,FⅢ 2 (4 ,4 0或 4 0 0mg·L- 1)显著提高低剂量脂多糖 (LPS) 10mg·L- 1协同佛波醇 14烷酰乙酸盐 (PMA) 2 0 0nmo1·L- 1诱导的人组织瘤THP 1细胞产生肿瘤坏死因子α(TNFα) ,然而明显地抑制高剂量LPSl0 0mg·L- 1协同PMA诱导的THP 1细胞产生TNFα .FⅢ 2 S(4或4 0mg·L- 1)提高无刺激剂细胞产生TNFα ,高浓度FⅢ 2 S(4 0 0mg·L- 1)抑制TNFα产生 .FⅢ 2与FⅢ 2 S提高白介素 8的产生 ,降低刺激剂引起的白介素 1α的产生 .FⅢ 2的抗肿瘤作用可能是与TNFα产生有关 .FⅢ 2对人外周血单核细胞产生各种细胞因子和对THP 1细胞产生细胞因子的机理基本上相同 .结果提示 ,松杉灵芝菌丝体水不溶性多糖在不同的刺激条件下具有双向免疫调节作用 .化学修饰的多糖可改变原多糖对细胞因子产生的调节方向 .     

Development of the "Cell Chip": a new in vitro alternative technique for immunotoxicity testing

Predictive testing of immunotoxicity associated with chemical compounds is complicated and cannot be accomplished with a single test. As most of the existing tests for immunotoxicity employ experimental animals, there is an increasing need for alternative tests in vitro. We have developed a new system for in vitro immunotoxicity testing, which employs changes in cytokine expression observed in vitro as an endpoint indicating potential for perturbation of the immune system in vivo. This system named "fluorescent cell chip" (FCC) is based on a number of genetically modified cell lines that regulate the expression of a transgene coding for fluorescent protein enhanced green fluorescent protein (EGFP) in a similar way as they regulate expression of IL-1beta, IL-2, IL-4, IFN-gamma, IL-10, TNF-alpha, and beta-actin. Morphological and functional features of selected cell lines expressing EGFP under the control of cytokine promotors were compared with maternal cell lines and this comparison showed that critical functional features of the maternal cell lines were preserved in EGFP expressing cells. Two chemicals with known immunotoxic activities, cyclosporine A and potassium tetrachloro-platinate(II), mediated compound-specific pattern of inhibition and activation of reporter gene expression. Thus, the "fluorescent cell chip" has demonstrated potential for application as a predictive screening test for immunomodulatory activities of chemicals. The major advantage of this approach is the possibility to apply this test in high throughput screening of high number of compounds for their well defined biological activity.     

High glucose increases the expression of proinflammatory cytokines and secretion of TNFα and β-hexosaminidase in human mast cells

Epidemiological studies demonstrated that obesity, which is a high-risk factor for development of hyperglycemia-associated metabolic syndromes, is associated with prevalence/incidence of allergic diseases. To elucidate the underlying mechanisms of the relationship between hyperglycemia and allergy, we examined the effect of high glucose on the activation of human mast cell lines, HMC-1 and LAD2. HMC-1 and LAD2 cells were cultured in low (5.5 mM) and high (25 mM)-glucose Dulbecco's modified Eagle's medium (DMEM). High-glucose medium increased the intracellular reactive oxygen species levels in HMC-1 and LAD2 cells after 2 days of incubation; in HMC-1 cells, the expression levels of tumor necrosis factor (TNF) α, interleukin (IL)-1β, IL-6, and IL-13 were increased significantly. The β-hexosaminidase release rates were not significantly different between LAD2 cells cultured in both media; however, the intracellular and extracellular activities of β-hexosaminidase in cells were significantly higher in high-glucose than in low-glucose media. High glucose increased the secretion of TNFα by unstimulated HMC-1 cells and IgE crosslinking-stimulated LAD2 cells. High glucose increased the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinases (MAPKs), which regulate the expression of TNFα and other inflammatory cytokines, in both HMC-1 and LAD2 cells. Thus, high glucose increased the expression of proinflammatory and proallergic cytokines, the secretion of TNFα, and β-hexosaminidase activity in human mast cells. Our result suggests that hyperglycemia promotes the activation of human mast cells associated with allergy and inflammation under unstimulated and stimulated conditions.     

Immune stimulating properties of a novel polysaccharide from the medicinal plant Tinospora cordifolia

An alpha-D-glucan (RR1) composed of (1-->4) linked back bone and (1-->6) linked branches with a molecular mass of >550 kDa and exhibiting unique immune stimulating properties is isolated and characterized from the medicinal plant Tinospora cordifolia. This novel polysaccharide is noncytotoxic and nonproliferating to normal lymphocytes as well as tumor cell lines at 0-1000 microg/ml. It activated different subsets of the lymphocytes such as natural killer (NK) cells (331%), T cells (102%), and B cells (39%) at 100 microg/ml concentration. The significant activation of NK cells is associated with the dose-dependent killing of tumor cells by activated normal lymphocytes in a functional assay. Immune activation by RR1 in normal lymphocytes elicited the synthesis of interleukin (IL)-1beta (1080 pg/ml), IL-6 (21,833 pg/ml), IL-12 p70 (50.19 pg/ml), IL-12 p40 (918.23 pg/ml), IL-18 (27.47 pg/ml), IFN- gamma (90.16 pg/ml), tumor necrosis factor (TNF)-alpha (2225 pg/ml) and monocyte chemoattractant protein (MCP)-1 (2307 pg/ml) at 100 microg/ml concentration, while it did not induce the production of IL-2, IL-4, IL-10, interferon (IFN)-alpha and TNF-beta. The cytokine profile clearly demonstrates the Th1 pathway of T helper cell differentiation essential for cell mediated immunity, with a self-regulatory mechanism for the control of its overproduction. RR1 also activated the complements in the alternate pathway, demonstrated by a stepwise increase in C3a des Arg components. Incidentally, RR1 stimulation did not produce any oxidative stress or inducible nitric oxide synthase (iNOS) in the lymphocytes or any significant increase in nitric oxide production. The water solubility, high molecular mass, activation of lymphocytes especially NK cells, complement activation, Th1 pathway-associated cytokine profile, together with a low level of nitric oxide synthesis and absence of oxidative stress confer important immunoprotective potential to this novel alpha-D-glucan.     

Effect of interleukin-1-beta and tumor necrosis factor-alpha on cartilage proteoglycan metabolism in vitro

The activities of recombinant interleukin-1-beta (IL-1) and recombinant tumor necrosis factor-alpha (TNF) on cartilage proteoglycan metabolism were compared in an organ culture system. IL-1, 1 to 100 ng/ml, and TNF, 10 to 1,000 ng/ml, increased proteoglycan degradation. The concentration-response curves were parallel. The timecourse for degradation was similar for the two cytokines during a 6 day incubation. Both cytokines inhibited the synthesis of new proteoglycan as measured by 35S incorporation. The inhibition curves were parallel and concentration-related between 1 and 10 ng/ml for IL-1 and between 10 and 100 ng/ml for TNF. Maximal inhibition was 60% in the presence of IL-1 (10 ng/ml) or TNF (100 ng/ml), and plateaued at higher concentrations. IL-1 was ten fold more potent than TNF in stimulating proteoglycan breakdown and inhibiting proteoglycan synthesis. Degradation in response to TNF, but not to IL-1, could be blocked by a polyclonal antibody to TNF. A polyclonal antibody to IL-1 could block proteoglycan breakdown in response to both cytokines suggesting that TNF may be mediating proteoglycan degradation by inducing the production of interleukin-1.     

Effect of corticosteroids,cyclosporin A,and methotrexate on cytokine release from monocytes and T-cell subsets

Corticosteroids are the most effective drugs in the management of asthma. However, because of their known side effects and the existence of corticosteroid-resistant patients, there is a need for substitute medications in asthma therapy. Using cell lines, in the present study, the two corticosteroids dexamethasone (Dex), and beclomethasone (Bec), as well as the immunosuppressant cyclosporin A (CsA), and the antimetabolic drug methotrexate (Mtx) were examined in their effect on release of immunoreactive IL-1β, IL-2, IL-4, IL-5, and IL-8. THP-1 cells served as a test model for monocytes secreting IL-1β and IL-8 upon stimulation by lipopolysaccharide. Jurkat cells were used as a test model for T_H 1-type T-cells and were stimulated for IL-2 release with a combination of phytohemagglutinin and phorbol myristate acetate. Representing T_H2-type T-cells, D10.G4.1 cells challenged by anti-CD3-mAb produced IL-4, and IL-5. Considerable qualitative and quantitative differences in the relative efficacy of the test compounds were found. Following IC50 values (nmol/1) of the test compounds were estimated (IL-1β/IL-8/IL-2/IL-4/IL-5): Dex (10.8/35.7/>10,000.0/5.1/4.1), Bec (30.9/102.2/8591.4/0.6/0.4), CsA (318.7/6211.2/2.3/68.2/237.9). Mtx in concentrations up to 10,000.0 nmol/1 was completely inactive. It can be concluded that corticosteroids show another inhibition pattern than CsA: corticosteroids affect mainly T_h2-type T-cells, while CsA primarily inhibits the T_h1-type T-cell response.)