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91.
Idziorek Khalife Billaut-Mulot Hermann Aumercier Mouton Capron Bahr 《Clinical and experimental immunology》1998,112(1):84-91
The chemoattractant cytokine IL-16 has been reported to suppress lymphocyte activation and to inhibit HIV-1 replication in acutely infected T cells. We have cloned and expressed human IL-16 in Escherichia coli and investigated whether the recombinant protein could regulate the level of lymphocyte apoptosis from HIV-1-infected subjects. After purification and refolding, only 2–10% of the recombinant cytokine was present in a biologically active homotetrameric form. This could explain the need for high concentrations of the bacterially derived IL-16 to induce significant inhibition of HIV-1 replication. Addition of IL-16 to unstimulated peripheral blood mononuclear cell (PBMC) cultures from HIV-1-infected subjects did not modify the observed level of spontaneous lymphocyte apoptosis. In contrast, IL-16 added to PBMC cultures stimulated with anti-CD3, anti-CD95 or dexamethasone reduced significantly the percentage of lymphocytes undergoing AICD. This effect was found to correlate with the ability of the cytokine to decrease CD95 expression on activated CD4+ T cells. Comparative studies on PBMC from healthy individuals indicated that the regulation of apoptosis levels by IL-16 is a complex phenomenon and could depend on the nature of the activator used and/or the immune status of lymphocytes tested. The outcome of CD4 cross-linking on T cells by various ligands is discussed in the context of the observed beneficial activities of IL-16 and its potential role in the treatment of HIV disease. 相似文献
92.
A. Sklavounou-Andrikopoulou E. Chrysomali M. Iakovou G. A. Garinis A. Karameris 《Journal of oral pathology & medicine》2004,33(7):386-390
BACKGROUND: The serum circulatory levels of apoptosis related molecules measured in patients with oral lichen planus (OLP) and healthy individuals in order to investigate possible alterations associated with the clinical forms of OLP. METHODS: Serum levels of tumor necrosis factor (TNF)-alpha, soluble Fas (sFas) and Bcl-2 studied by enzyme-linked immunosorbent assay in whole blood samples in 13 OLP reticular, 13 OLP atrophic-erosive form patients and 26 healthy subjects. RESULTS: Significantly elevated levels of TNF-alpha and sFas detected in OLP patients as compared with controls. Serum concentrations of Bcl-2 although increased in 17/26 patients, they were not statistically significant. Reticular OLP exhibited slightly elevated TNF-alpha and significantly elevated Bcl-2 serum levels, compared with erosive OLP. CONCLUSIONS: These data suggest that a putative dysfunction in the Fas/FasL mediated apoptosis might be involved in the OLP pathogenesis. A downregulation of Bcl-2 serum levels in the atrophic-erosive OLP may be associated with promotion of the disease activity. 相似文献
93.
凋亡相关蛋白Apr-2的克隆、测序及初步表达 总被引:1,自引:0,他引:1
目的: 从HL-60细胞凋亡模型中克隆凋亡相关蛋白Apr-2编码区基因,并对其进行表达,为进一步研究Apr-2的结构功能及多克隆抗体的制备奠定基础.方法: 建立HL-60细胞凋亡模型,提取HL-60凋亡细胞总RNA,以RT-PCR方法获取Apr-2 cDNA编码区全序列,将其与PGEM-T Easy载体连接,转化E.coli DH5α,构建重组克隆载体PGEM-TEasy/apr-2,测序正确后,将目的片段亚克隆入PGEX-4T-2原核表达载体,并转化大肠杆菌, IPTG诱导重组蛋白质表达,分析蛋白质在细菌中的表达分布,进行凝胶自动扫描分析. 结果:序列分析表明,与GenBank中已登录的Apr-2 cDNA编码区序列比较,完全一致.表达的融合蛋白占菌体总蛋白质的40%以上,主要以包涵体的形式存在. 结论:成功的获得了细胞凋亡模型HL-60中Apr-2 cDNA编码区的克隆及其融合蛋白表达产物. 相似文献
94.
目的 :研究氯化锂 (LiCl)在体外对K5 6 2白血病细胞增殖及凋亡的影响。方法 :采用液体培养试验 ,四唑盐 (MTT)比色试验 ,集落培养试验为指标观察LiCl对K5 6 2细胞增殖的影响 ,采用DNA片段凝胶电泳及细胞形态学改变为指标检测细胞凋亡。结果 :①不同浓度的LiCl(10~ 5 0mmol/L)对K5 6 2细胞具有抑制增殖的作用 ,这种增殖抑制作用呈剂量依赖关系。②在LiCl(30mmol/L)作用下 ,K5 6 2细胞形态学上可见凋亡细胞 ,且DNA琼脂糖凝胶电泳谱可见“梯形带”(DNAladder)。结论 :LiCl能抑制K5 6 2白血病细胞增殖和诱导其凋亡。 相似文献
95.
甲基强的松龙治疗实验性变态反应性脑脊髓炎的作用机制 总被引:1,自引:0,他引:1
目的:研究细胞因子、T细胞凋亡和淋巴细胞增殖在实验性变态反应性脑脊髓炎(EAE)形成中的作用及甲基强的松龙(MP)治疗EAE的作用机制。方法:采用人脑纯化的髓鞘碱性蛋白(MBP)与完全福氏佐剂免疫Lewis大鼠,建立EAE动物模型。用双抗体夹心ELISA法检测各组大鼠血清中IL-10、TNF-α、IFN-γ的含量:流式细胞仪检测外周血T细胞凋亡;3H-TdR释放法检测外周血淋巴细胞转化率。结果:与对照组比较,EAE组的外周血IFN-γ、TNF-α水平明显增高,IL-10水平明显降低,MP治疗后IFN-γ和TNF-α水平下降,IL-10浓度上调。MP还诱导外周血T细胞凋亡和抑制MBP致敏淋巴细胞增殖并呈剂量依赖性。结论:应用人MBP成功建立EAE大鼠模型,MP可能通过调节Th细胞因子格局、促进Th2细胞因子分泌、抑制MBP致敏淋巴细胞增殖及外周血T细胞凋亡而发挥治疗多发性硬化的作用。 相似文献
96.
TETSUO HAYASHI SOICHIRO YOSHIDA ATSUSHI YOSHINAGA RENA OHNO NOBUYUKI ISHII TAKUMI YAMADA 《International journal of urology》2006,13(2):157-164
AIM: The aim of the present study was to elucidate the role of high temperature requirement A2 (HtrA2) in germ cell loss in the heat-stressed testis. METHODS: We examined the expression of HtrA2, caspase-9 activity and proteolytic activity of HtrA2 in the rat testis, and their in vivo responses to experimental cryptorchid treatment. RESULTS: Northern analysis revealed the expression of HtrA2 mRNA peaked at days 1 and 7 after cryptorchid treatment. While expression of HtrA2 mRNA was seen in the spermatogonium, spermatocytes and some spermatids in normal adult rat testis, experimental cryptorchidism treatment resulted in a marked increase in its signal intensity in spermatocytes and some spermatids, and the layers of spermatogonium and early primary spermatocytes became negative at days 1 and 7 after the treatment. However, the spermatogonium, Sertoli cells and interstitial cells appeared to have strong intensities at days 14, 28 and 56 after the treatment. Western analysis revealed the expression of HtrA2 protein peaked at day 2 coinciding with the increase of positive spermatogonium, the appearance of protein-positive interstitial cells, and day 28 coinciding with the reappearance of protein-positive interstitial cells. Caspase-9 activity peaked at day 2 and HtrA2 proteolytic activity peaked at day 28. Consequently, the first peak of HtrA2 mRNA expression was followed by the peak of caspase-9 activity and the second peak was followed by the peak of proteolytic activity; however, the second peak of mRNA expression had considerable chronological difference from that of the protein. CONCLUSION: These findings suggest the probabilities that the heat stress results in germ cell death by a caspase-independent manner with the elevation of HtrA2 proteolytic activity, as well as a caspase-dependent manner with the elevation of caspase-9 activity. 相似文献
97.
幽门螺杆菌感染对胃黏膜上皮细胞端粒酶活性的影响及其与细胞凋亡的关系 总被引:2,自引:0,他引:2
目的探讨幽门螺杆菌(Hp)感染对端粒酶活性的影响及其与细胞凋亡的关系。方法慢性胃炎伴萎缩者16例,伴肠化生者15例,伴中、重度不典型增生者14例,胃癌19例共64例为研究对象。采用快速尿素酶试验、Warthin-Starry银染色检测幽门螺杆菌,采用S-P法检测人端粒酶催化亚单位(hTERT)蛋白表达,采用TUNEL染色法原位检测细胞凋亡。结果(1)萎缩性胃炎、肠化生、不典型增生和胃癌组织中hTERT蛋白表达率分别为25.00%、46.67%、64.29%和78.95%,呈递增趋势。Hp阳性患者hTERT蛋白表达率为77.42%,显著高于Hp阴性患者(33.33%,P<0.01)。(2)Hp阳性伴hTERT蛋白阳性患者细胞凋亡指数为9.57±5.01,显著高于Hp阴性伴hTERT蛋白阳性患者(6.01±5.31,P<0.05)。结论(1)端粒酶激活可能参与了胃黏膜上皮细胞恶性转化的全过程。Hp感染与端粒酶活性密切相关。Hp感染可能是激活端粒酶的重要机制之一。(2)Hp感染既可激活端粒酶,引发细胞永生化,又可以诱导胃黏膜上皮细胞凋亡,进而增加了胃黏膜的不稳定性和癌变的危险性,这可能是Hp致癌的主要机制之一。 相似文献
98.
目的 探讨维生素E琥珀酸酯 (VES)诱导人胃腺癌SGC -790 1细胞凋亡的死亡受体 (Fas)信号转导途径。方法 人胃腺癌SGC -790 1细胞经不同剂量VES(5,10 ,2 0 μg/ml)处理 ,同时做琥珀酸、维生素E和空白对照 ,采用DAPI(4,6-贰脒基 -2 -苯基吲哚 )荧光染色法观察细胞凋亡情况 ,用WesternBlot法检测Fas、带有死亡结构域的Fas相关蛋白 (FADD)和天冬氨酸特异性半胱氨酸蛋白酶 (caspase -8)蛋白表达情况 ,Fas和FADD反义寡聚核苷分别转染SGC -790 1细胞后 ,用荧光法检测caspase -8活性。 结果 经VES处理后的细胞DAPI染色可见凋亡的形态学改变 ,2 0 μg/mlVES处理 48h后的细胞凋亡率为 89.6% ;VES处理 48h后Fas、FADD和caspase -8蛋白表达明显增加 ,且呈剂量 -效应关系 ;阻断Fas可明显抑制FADD蛋白表达 ,Fas和FADD反义寡聚核苷转染细胞后caspase -8活性明显降低 (P <0 .0 1) ,其中阻断Fas的效果高于阻断FADD。结论 维生素E琥珀酸酯诱导人胃腺癌SGC -790 1细胞凋亡过程中启动了Fas信号转导途径 ,VES启动Fas后 ,FADD将Fas和caspase -8联接起来 ,活化caspases级联反应 ,从而构成Fas/FADD/caspase -8的凋亡信号途径 相似文献
99.
Liver biopsy remains the 'gold standard' for monitoring rejection in liver transplant patients. Portal inflammation, bile duct damage and endothelialitis are recognized features of hepatic allograft rejection. The pathogenesis of the bile duct injury during rejection, however, remains unclear. To define the mechanism of bile duct damage, we studied the light- and electronmicroscopic appearance of hepatic tissue from selected patients in whom allograft failure was solely due to rejection. Of the 25 orthotopic liver transplant rejection cases examined, 17 were mild, seven were moderate and one was severe rejection. Light microscopy examination of the damaged bile duct epithelium revealed evidence of apoptosis which was confirmed by electronmicroscopy. Furthermore, there appeared to be a positive correlation between the grade of rejection and the number of apoptotic cells. Also included in the study were 13 cases of chronic active hepatitis and 10 normal livers which showed the least apoptotic cells. We conclude that the identification of apoptotic cells in damaged bile ducts in allograft biopsies might be helpful in the diagnosis of rejection and in assessment of the severity of rejection. 相似文献
100.
Werner I Hagens Peter Olinga Dirk K F Meijer Geny M M Groothuis Leonie Beljaars Klaas Poelstra 《Liver international》2006,26(2):232-239
BACKGROUND: Liver fibrosis is the common response to chronic liver injury, ultimately leading to cirrhosis. Several lines of evidence indicate that inducing apoptosis of hepatic stellate cells (HSC) may lead to regression of liver fibrosis. Recently, it was shown that gliotoxin (GTX) induces apoptosis of HSC. However, the clinical use of GTX may be limited because of the lack of cell and tissue specificity, causing a high risk of potentially severe adverse effects. The aim of this study, therefore, was to study the effect of GTX on different cells of the liver. METHODS: We used normal and fibrotic precision-cut rat liver slices to study the effect of GTX on the various resident liver cell types. In these slices, the complex cell-cell interactions are preserved, which closely mimics the in vivo situation. RESULTS: GTX exhibited a potent apoptosis-inducing activity in these slices. Both immunohistochemical stainings and real-time mRNA techniques showed that this apoptosis-inducing effect was seen in HSC. However, Kupffer cells and liver endothelial cells were also affected by GTX, whereas hepatocytes were only mildly affected. CONCLUSIONS: This study indicates that the apoptosis-inducing strategy to treat liver fibrosis has high potential, but it will be necessary to develop an HSC-specific therapy to prevent adverse effects. 相似文献