The effectors of killer activity induced by interferon-alpha and gamma in peripheral blood mononuclear cells

The nature of effectors of interferon (IFN)-alpha or IFN-gamma-induced killer cell activity remains unclear. The aim of this study was to examine killer cell activity induced by IFN-alpha alone, IFN-gamma alone or a combination of both in patients with renal cell carcinoma (RCC) and to determine the phenotypic patterns of these effectors. The study group included 14 patients (12 men and 2 women, median age 64 years, range 36-77) with confirmed RCC. Peripheral blood mononuclear cells (PBMC) from RCC patients or normal volunteers were cultured with IFN-alpha alone, IFN-gamma alone or a combination of both. Cytotoxic activity was assayed against ACHN cells. Subpopulations of effector cells in IFN-induced killer cell activity were characterized by cell sorting. The most effective type of IFN and the optimal concentration of IFN necessary to induce the maximal killer cell activity varied among RCC patients. The killer activity induced by a combination of IFN-alpha and IFN-gamma was significantly greater than that induced by IFN-alpha or IFN-gamma alone. The greatly increased killer activity induced by IFN-alpha and IFN-gamma was seen in the subpopulations CD3(-) CD16(+), CD3(-) CD56(+) and subpopulation CD3(+)CD4(-), CD3(-)CD16(+), CD3(-)CD56(+), CD57(+)CD16(-), respectively. An optimal type of IFN and optimal concentration of IFN seem to increase the effective rate of treatment of RCC. In addition, the role of IFN-alpha seems to be different from that of IFN-gamma in host defense against RCC. A combination treatment with IFN-alpha and IFN-gamma seems to be suitable to increase the effective rate if we could reduce the side effects of IFNs.     

启动子区5′CpG岛去甲基化对人结肠癌细胞生物学表型的影响

目的：探讨DNA启动子区5′CpG岛甲基化状态与人结肠癌RKO细胞增殖凋亡等生物学特征的关系。方法： 应用特异性DNA甲基转移酶（DNMTs）抑制剂-5-氮-2′-脱氧胞苷（5-Aza-2′-deoxycytidine，5-Aza-CdR）处理肠癌RKO细胞72 h，甲基化特异性PCR（methylation-specific PCR,MSP）及DNA测序法分析p16/CDKN2基因CpG岛甲基化状态；MTT、FCM、荧光染色及透射电镜检测启动子区去甲基化后细胞生长、形态和细胞周期凋亡的影响。 结果： DNMTs抑制剂能较好地逆转启动子区胞嘧啶甲基化状态；CpG岛去甲基化后能明显地抑制肠癌细胞的生长，增加细胞群体倍增时间（P<0.01），诱导肠癌细胞凋亡，影响肠癌细胞周期分布，并具有良好的量效依赖关系。 结论： 通过逆转CpG岛高甲基化能有效地抑制肠癌细胞增殖，为临床治疗大肠癌提供新的作用靶点。     

Interferon-{beta} inhibits bleomycin-induced lung fibrosis by decreasing transforming growth factor-{beta} and thrombospondin

Pulmonary fibrosis is the result of abnormal processes of repair that occur after lung injury. Transforming growth factor (TGF)-beta is a key molecule in the progression of pulmonary fibrosis. Although clinical use of interferon (IFN)-beta did not improve survival in patients with idiopathic pulmonary fibrosis, because some preclinical studies have suggested that IFN-beta is a potent inhibitor of fibrogenesis, beneficial effects of IFN-beta have been expected. We therefore attempted to determine effects of IFN-beta and investigated the mechanism of action of IFN-beta in bleomycin-induced pulmonary fibrosis. Bleomycin at Day 0 and IFN-beta for 4 wk were administered intravenously to ICR mice. At 28 d after bleomycin injection, histologic and chemical analysis was performed for evaluation of effects of IFN-beta. Tissue distribution and amounts of TGF-beta1 and thrombospondin (TSP)-1/2 were analyzed. IFN-beta attenuated prolylhydroxylase activity, resulting in inhibition of pulmonary fibrosis. Bleomycin-induced increase in TGF-beta1 in epithelial cells and extracellular matrix was attenuated by IFN-beta. TSP-1/2 was limited in platelets of control mice, but was present in foamy cells in fibrotic regions induced by bleomycin. These findings suggest that the antifibrotic effect of IFN-beta is inhibition of TGF-beta and its activation via decrease in TSP-1/2 in lung tissue and change in location of TSP-1/2 from platelets to foamy cells.     

Induktion von endogenem Pyrogen und Interferon durch Newcastle Disease-Virus in vivo

Zusammenfassung Die Ausbildung von endogenem Pyrogen und Interferon wurde nach der Injektion von NDV im Kaninchen untersucht. Das Auftreten und das Verschwinden beider Substanzen stimmte nicht nur zeitlich, sondern auch quantitativ überein. Zudem verliefen die Bildungskurven von endogenem Pyrogen und Interferon sowohl im Blut als auch in den Organen weitgehend gleichsinnig. Eine Abweichung von diesem Verhalten wurde lediglich in der Milz beobachtet, indem hier das endogene Pyrogen bereits nach 1 Stunde, das Interferon jedoch erst nach 3 Stunden den höchsten Gipfel erreichte. Dieser Befund deutet daraufhin, daß das endogene Pyrogen und das Interferon zwei verschiedene Substanzen sind.Die Exstirpation der Milz hatte ein gleichzeitiges Absinken des endogenen Pyrogens und des Interferons im Blut, jedoch nicht in der Leber, der Lunge und der Niere zur Folge. Hinsichtlich der Entstehung der partiellen bzw. kompletten Toleranz gegenüber dem NDV bzw. Influenza Virus A (PR8) dürften die beiden Substanzen demselben Mechanismus unterliegen.

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Induction of endogenous pyrogen and interferon by newcastle disease virus in vivo

Summary Endogenous pyrogen and interferon induced by injection of rabbits with NDV were investigated. The appearance and disappearance of both substances were in parallel in time as well as in quantity. The curves of endogenous pyrogen and interferon in the blood and in various organs tested usually were also parallel. Only in spleen highest titers of endogenous pyrogen were found already 1 hour after inoculation, whereas interferon activity reached its peak only after 3 hours. This finding indicated that endogenous pyrogen and interferon may be different substances.In splenectomized animals the blood levels of endogenous pyrogen and interferon were lower than those found in intact animals, while the titers in liver, lung and kidneys were identical. In respect to the formation of partial or complete tolerance to NDV and influenza virus A (PR8), respectively, both substances appear to be subjected to the same mechanism.

     

CTLA-4-FasL融合分子的构建及其生物学特性的鉴定

文章通过特异的引物分别扩增出CTLA 4和FasL胞外区的cDNA ,将它们拼接后 ,克隆入真核表达载体pcDNA3 1( + )中 ,进行表达、纯化 ,获得CTLA 4 FasL融合蛋白。Westernblot分析显示了该融合蛋白具有CTLA4 胞外区和FasL胞外区的抗原性。体外试验表明 ,该融合蛋白可以结合Jurkat细胞表面的Fas受体和Raji细胞表面的B7分子 ,表明了该分子双特异性的特点。该融合蛋白能够直接诱导Jurkat细胞发生凋亡 ,且此凋亡效应伴随Raji细胞的参与而增强 ,初步证实了该分子的免疫抑制效应 ,从而为进一步研究该融合蛋白特性及应用奠定了基础。     

小鼠β_2m反义核酸重组荧光蛋白表达载体的构建及表达研究

本研究应用分子生物学技术 ,构建小鼠 β2 m反义核酸重组荧光蛋白表达载体pEGFP β2 mAN ,经过酶切、PCR鉴定 ,以及序列分析证实克隆的正确性。然后用脂质体转染NIH3T3细胞 ,经过RT PCR及Westernblot检测 ,观察重组质粒对细胞β2 mmRNA和蛋白表达的影响 ,同时在荧光显微镜下直接观察细胞转染的结果。结果证明pEGFP β2 mAN已成功导入NIH3T3细胞中 ,且有效表达 ,在荧光显微镜下可见梭形绿色荧光细胞 ;与同步转染的小鼠 β2 m正、反义核酸表达载体pcDNA3 β2 mSN、pcDNA3 β2 mAN的转染结果一起进行分析 ,转染pcDNA3 β2 mSN细胞的 β2 mmRNA和蛋白表达水平升高 ,而转染pcD NA3 β2 mAN和pEGFP β2 mAN的细胞 β2 mmRNA和蛋白表达水平降低。小鼠 β2 m反义核酸重组荧光蛋白表达载体pEGFP β2 mAN的转染结果显示 :它不但可以降低NIH3T3细胞 β2 m基因的表达 ,而且为观察脂质体转染效果提供了直观、即时的方法     

用电子自旋共振成像观测氮氧自由基的透皮特性

在自行研制的电子自旋共振成像(Electron spin resonance imaging, ESRI)系统上建立了活体皮肤电子自旋共振成像的实验模型.观测了氮氧自由基化合物2,2,6,6-四甲基哌啶-1-氧(2,2,6,6-tetramethyl-1-piperidinyloxy,TEMPO)穿透小鼠皮肤组织的动态过程和浓度分布情况.成像结果显示TEMPO化合物对不同方法处理后的皮肤组织的透皮特性有明显差别.TEMPO水溶液难以穿透皮肤,而溶解在二甲基亚枫(Dimethyl sulfoxide,DMSO)中的脂溶性TEMPO溶液可以渗入并穿透皮肤.皮肤表层的角质蛋白对TEMPO有明显的阻隔性, 而且化合物渗入或透过皮肤组织的量具有较强的时间依赖性.本研究结果提示ESRI技术为动态研究某些特殊结构化合物的透皮特性提供了科学有效的分析手段.     

阿糖胞苷纳米粒的制备及其释药规律研究

采用乳化聚合法制备阿糖胞苷纳米粒，研究其体内外释药特性。结果表明阿糖胞苷纳米粒体外释药规律符合双指数方程，有明显的缓释作用。在家兔体内的药物动力学过程符合二室模型，与阿糖胞苷注射剂相比，t1／2β和MRT延长，CL降低，表明阿糖胞苷纳米粒可显著延长阿糖胞苷在体内存留时间，具有明显的缓释特征。     

大鼠肝移植排斥反应期一氧化氮合酶的动力学变化

目的 探讨供体特异性输血（DST）预处理后大鼠同种肝移植物一氧化氮合酶（NOS）的动力学变化。方法 观察同基因肝移植（Ⅰ组），同种肝移植（Ⅱ组），DST预处理的同种肝移植（Ⅲ组）术后NOS的表型及亚硝酸盐/硝酸盐，细胞因子的动态变化。结果 Ⅱ组血中亚硝酸盐/硝酸盐，INF-α，TNF-α浓度明显增高，免疫组化显示，Ⅱ组移植物中macNOS^ 及ED1^ ,ED2^ 细胞数明显增多，Northern blot分析，Ⅲ组移植物中IL-10和TGF-BmRNA呈高水平表达；Ⅱ组移植物的iNOS和IL-12mRNA水平明显高于其他组。结论 DST预处理的移植物处于免疫无应答状态，NOS被抑制与TH2及TH3类细胞因子高表达有关。