WEHI164—C1细胞株检测TNF的MTS／PMS比色法的建立

本文用有限稀释法对ＷＥＨＩ１６４细胞进行进行了亚克隆，从１２株中筛选出一株对ＴＮＦ高度敏感的亚克隆细胞株ＷＥＨＩ－Ｃｌ０ＭＴＳ的还原产物具有良好的水溶性，ＭＴＳ／ＰＭＳ比色法与ＭＴＴ法相比较具有简便、快速的优点。我们首先将ＭＴＳ引入细胞毒的检测，用ＷＥＨＩ１６４－Ｃ１亚克隆株建立了检测ＴＮＦ的ＭＴＳ／ＰＭＳ比色法。此方法不仅简便、快速，而且敏感度高（比常规Ｌ９２９细胞检测法敏感１０－１５倍）、特异     

一种快速、简便、敏感、特异的白细胞介素2、3、6生物学检测法：MTS／PMS法的建立

ＭＴＴ的一种新型类似物ＭＴＳ在ＰＭＳ存在情况下可被活细胞还原，形成水溶性甲产物。本实验表明，将ＭＴＳ／ＰＭＳ取代ＭＴＴ用于ＩＬ－２、３、６生物学测定法取得理想结果，ＭＴＳ／ＰＭ５形成的比色产物与细胞数量、ＭＴＳ／ＰＭＳ与细胞孵育时间呈正相关。ＭＴＳ／ＰＭＳ生物学测定法省略了加有机溶剂溶解甲的步骤，ＭＴＳ／ＰＭ５贮存稳定、显色快速，故该法具有快速、简便敏感、特异的优点，适用于细胞因子的生物学测定。     

A murine transmembrane tumor necrosis factor (TNF) transgene induces arthritis by cooperative p55/p75 TNF receptor signaling

The arthritogenic activities of tumor necrosis factor (TNF) and its p55TNF-receptor (R) have been well documented in experimental animal models of arthritis, and in transgenic mice expressing wild-type or mutant transmembrane human TNF proteins in their joints. In this study we show that chronic inflammatory arthritis also develops in transgenic mice made to overexpress a mutant transmembrane from of the murine TNF protein (muTNF_Δ1–12) which is known to utilize efficiently both the p55 and the p75TNFR. Cross-breeding of the transgene into a TNF knockout background did not alter development of disease. Analysis of TNF bioactivity in sera from lipopolysaccharide-stimulated mice or ex vivo macrophage cultures demonstrated that the muTNF_Δ1–12 protein accumulates on the cell surface and is not processed to bioactive soluble TNF, indicating that transmembrane TNF is by itself sufficient to mediate pathogenesis of arthritis. Furthermore, using TNFR knockout mice, it is shown that development of transmembrane TNF-mediated arthritis requires the presence of the p55TNFR but is significantly delayed in the absence of the p75TNFR, suggesting a positive cooperation between the two TNFR in the arthritogenic process. These results indicate that blocking the activities of both soluble and transmembrane TNF may be required to effectively neutralize the pathogenic potential of this cytokine in arthritis.     

The type I interleukin-1 receptor acts in series with tumor necrosis factor (TNF) to induce arthritis in TNF-transgenic mice

The pro-inflammatory cytokines tumor necrosis factor (TNF) and interleukin-1 (IL-1) have been shown to be primary mediators in the pathogenesis of chronic inflammatory joint diseases. However, the relative contributions of these molecules to the development and progression of disease is not known. In the present study, we have investigated the involvement of the type I IL-1 receptor in the development and progression of chronic arthritis in a previously described TNF-transgenic mouse model of this disease. A neutralizing monoclonal antibody to the murine type I IL-1 receptor was injected intraperitoneally three times a week from birth to 9 weeks of age. In the positive control group of untreated transgenic mice the incidence of arthritis was 100% after 9 weeks of age. The injection of normal hamster IgG did not influence the incidence or development of arthritis. In contrast, the injection of antibody to the type I IL-1 receptor completely prevented the development of arthritis. Clinical evaluation of disease was confirmed histologically, anti-receptor antibody-treated mice showed no sign of arthritic change. Moreover, the analysis of sera taken at the end of the study period showed that the neutralization of arthritis by IL-1 receptor antibody treatment was accompanied by significantly lower levels of circulating human TNF compared to the control groups and to untreated transgenic mice prior to disease onset. Taken together, these results show that IL-1 signaling blockade exerts a direct negative feedback effect on TNF production. Our results show that in TNF-transgenic mice the IL-1 receptor accepts the whole pathogenic load of TNF, thereby acting as a potent downstream mediator in the pathogenesis of chronic arthritis.     

Prevention of cyclophosphamide-induced diabetes in the NOD/WEHI mouse with deoxyspergualin

Ten out of 20 (50%) 17-week-old female NOD/WEHI mice developed an acute form of autoimmune diabetes when injected with two large doses of cyclophosphamide (CY), given at 14-day intervals. If these mice were treated under a prophylactic regimen with 2·5 mg/kg body weight per day of the novel immunosuppressant deoxyspergualin (DSP) the onset of diabetes was completely prevented. Moreover, DSP-treated animals showed reduced signs of pancreatic insulitis, had lower percentages of splenic lymphoid cells (SLC) expressing IL-2 receptors and Ly-6C antigens on their surfaces, and these cells released lower amounts of interferon-gamma (IFN) when stimulated in vitro. These data, providing evidence for the capacity of DSP to protect NOD/WEHI mice from experimental autoimmune diabetes and to modulate histo-immunological pathogenic pathways, indicate DSP as a drug of potential interest in the treatment of human insulin-dependent diabetes mellitus.     

In vivo evidence for a functional role of both tumor necrosis factor (TNF) receptors and transmembrane TNF in experimental hepatitis

The significance of tumor necrosis factor receptor 1 (TNFR1) for TNF function in vivo is well documented, whereas the role of TNFR2 so far remains obscure. In a model of concanavalin A (Con A)-induced, CD4⁺ T cell-dependent experimental hepatitis in mice, in which TNF is a central mediator of apoptotic and necrotic liver damage, we now provide evidence for an essential in vivo function of TNFR2 in this pathophysiological process. We demonstrate that a cooperation of TNFR1 and TNFR2 is required for hepatotoxicity as mice deficient of either receptor were resistant against Con A. A significant role of TNFR2 for Con A-induced hepatitis is also shown by the enhanced sensitivity of transgenic mice overexpressing the human TNFR2. The ligand for cytotoxic signaling via both TNF receptors is the precursor of soluble TNF, i.e. transmembrane TNF. Indeed, transmembrane TNF is sufficient to mediate hepatic damage, as transgenic mice deficient in wild-type soluble TNF but expressing a mutated nonsecretable form of TNF developed inflammatory liver disease.     

急性白血病患者血浆肿瘤坏死因子及抑制物

利用ＴＮＦ细胞毒生物学活性检测法和ＴＮＦ抑制物生物活性检测法，检测２２例初治急性白血病体内ＴＮＦ和ＴＮＦＩＮＨ水平。急性白血病患血浆ＴＮＦ水平明显增高。达１１．４２±６．０２ｕ／ｍｌ。抗人ＴＮＦａ单抗能完全中和Ｍ４，Ｍ５，Ｍ６型急性非淋巴细胞白血病患血浆ＴＮＦ活性。部分患血浆中同时存在ＴＮＦ和ＴＮＦＩＮＨ。ＴＮＦ阴性的患血浆中亦单独存在ＴＮＦＩＮＨ活性，和正常人相比明显增高，其对ｔｈＴＮＦ     

喉鳞状细胞癌MTS1／p16基因纯合子缺失的检测

为探讨MTS1／p16基因在喉癌中纯合子缺失情况及其与喉癌生物学行为的关系,本研究用比较PCR技术对59例喉鳞状细胞癌（LSCC）MTS1／p16基因纯合子缺失进行检测。剔除3例不可比样本,9／56 LSCC有MTS1／p16基因纯合子缺失,缺失率为16．07％。MTS1／p16基因纯合子缺失与LSCC病理分级无关;LSCC晚期（Ⅲ-Ⅳ）期的MTS1／p16基因纯合子缺失率（36．84％）明显高于早期（Ⅰ-Ⅱ期）肿瘤（5．4％）,P＜0．01,显示MTS1／p16基因纯合子缺失与LSCC的自然病程有关。认为MTS1／p16基因纯合子缺失是LSCC发生发展过程中的分子机制之一,可能与LSCC的恶性发展有关。     

实验性精索静脉曲张大鼠睾丸组织TNF-α和NO含量的变化

目的：研究精索静脉曲张（VC）对大鼠睾丸组织TNF-α和NO含量的影响。 方法： 将成熟雄性Wistar大鼠随机分为手术结扎（ VG）组，n=30；手术非结扎(SOG)组，n=20。分别测定两组大鼠睾丸组织中TNF-α和NO的含量，并加以比较。 结果： ①VG组左侧睾丸TNF-α、NO含量分别显著高于SOG组（P＜0.01)；②TNF-α与NO呈正相关（r=0.523, P＜0.01)。 结论: VC时睾丸TNF-α和NO的变化, 可能是造成睾丸损伤、精子生成障碍甚至不育的原因之一。     

老年矽肺结核患者血清肿瘤坏死因子水平的临床观察

为了解老年矽肺结核患者的免疫状况 ,采用放射免疫法检测了 6 9例老年矽肺结核患者的血清肿瘤坏死因子 (TNF)水平 ,并与老年肺结核患者和健康老年人做了比较。结果发现 :老年矽肺结核和老年肺结核患者的TNF水平 ,均显著低于健康老年人 (P <0 .0 1,P <0 .0 5 ) ,老年矽肺结核患者和老年肺结核患者血清TNF水平相差无显著性 (P >0 .0 5 )。TNF水平与痰菌无明显相关。老年肺结核患者和老年矽肺结核患者机体免疫力低下 ,在治疗中应加用免疫调节剂     

炎症刺激因子TNF-α诱导转录因子ESE-1在内皮细胞中的表达

目的: 探讨炎症刺激因子TNF-α介导炎症时上调血管内皮细胞Tie 2基因的机制。 方法: 采用RT-PCR、定量PCR以及Western blotting方法检测TNF-α作用于原代人主动脉内皮细胞株（HAECs）、原代人肺动脉内皮细胞株（HPAECs）时Tie 2基因及ETS转录因子mRNA及蛋白的表达。结果: TNF-α可增加血管内皮细胞Tie 2基因的表达。TNF-α不能增加内皮细胞HAECs和HPAECs的NERF-2、ELF表达；TNF-α刺激后约 24 h 第1次检测到ESE-1 mRNA表达。延长刺激作用时间后，发现在18-36h之间可测到ESE-1 mRNA表达，24 h 为高峰，而蛋白质表达在18 h极弱，36 h达峰值。检测中所观察到ESE-1表达在Tie 2表达之前且表达模式类似。结论: Tie 2基因在TNF-α刺激时的上调与NERF-2或ELF介导无关，ESE-1可能在炎症因子刺激后Tie 2的转录调节中起作用。     

大鼠出血性休克后肠源性内毒素血症及TNF,IL—1变化的动态观察

本实验采用大鼠出血性休克模型（４。００ｋＰａ，９０ｍｉｎ）动态观察了ＴＮＦ、ＩＬ－１等细胞因子的变化规律及其与肠源性内毒素血症的关系。研究发现：休克组动物休克后门、体循环均出现显著的内毒素血症，门脉系统血浆内毒素水平的变化趋与外周血ＴＮＦ一致，但其峰值早于后者。同是时，腹腔巨噬细胞ＩＬ－１在性在复苏后６～２４ｈ亦持续升高。休克治疗组给予多粘菌素Ｂ预防性治疗后，动物门、体循环内毒素含量均迅速下降，Ｔ