NF-κB和GM-CSF表达与乳腺癌骨转移相关性的研究

目的探讨核转录因子-κB（nuclear factor kappa-light-chain-enhancer of activated B cells,NF-κB）和粒-巨噬细胞集落刺激因子（granulocyte-macrophage clony-stimulating factor,GM-CSF）的表达与乳腺癌发生骨转移的相关性及其与临床病理指标的关系。方法采用免疫组化方法,检测52例乳腺癌骨转移患者、72例乳腺癌患者肿瘤组织中NF-κB和GM-CSF的表达。结果乳腺癌骨转移组NF-κB阳性表达率（69.2%）明显高于乳腺癌组（41.7%）,P=0.002;而乳腺癌骨转移组GM-CSF阳性表达率（34.6%）低于乳腺癌组（63.9%）,P=0.001;NF-κB和GM-CSF表达间无明显的相关性（P=0.490）;NF-κB表达与乳腺癌分子病理分型有相关性（P=0.000）,而GM-CSF表达与乳腺癌分子病理分型无明显相关性（P=0.991）;NF-κB表达与肿块大小、临床分期、淋巴结转移数目、受体状态、HER-2、p53表达有关,GM-CSF表达与其它临床病理指标均无相关性。结论 NF-κB表达与乳腺癌多项临床病理指标有关,其可能与乳腺癌骨转移发生有关。     

清肠栓对急性溃疡性结肠炎大鼠结肠黏膜GM—CSF及G-CSF的影响

目的：观察清肠栓对急性溃疡性结肠炎（UC）大鼠结肠黏膜粒-巨噬细胞集落刺激因子（GM—CSV）和粒细胞集落刺激因子（G—CSF）的影响。方法：48只SD大鼠随机分为正常组、模型组、清肠栓组和柳氮磺胺吡啶（SASP）组,每组12只。用三硝基苯磺酸（TNBs）复制大鼠急性溃疡性结肠炎模型。造模后第3天开始给药,连续给药7d后处死大鼠。采用免疫组化、实时荧光定量PCR检测结肠组织中GM—CSF、G—CSF蛋白及mRNA表达。结果：模型组GM-CSF和G-CSF蛋白表达较正常组升高（P〈0．05）,清肠栓组及SASP组均较模型组降低（P〈0．05）;模型组GM—CSF和G—CSF mRNA表达较正常组明显增高（P〈0．05）,清肠栓组及SASP组均较模型组减低（P〈0．05）。结论：清肠栓可通过调节结肠黏膜GM—CSF、G-CSF蛋白及基因表达,以治疗急性溃疡性结肠炎。     

Role of Paris PM2.5 components in the pro-inflammatory response induced in airway epithelial cells

Particulate matter (PM) is suspected to play a role in environmentally-induced pathologies. Due to its complex composition, the contribution of each PM components to PM-induced biological effects remains unclear.     

A synthetic analog of alpha-galactosylceramide induces macrophage activation via the TLR4-signaling pathways

Alpha-galactosylceramide (alpha-GalCer), a bioactive glycolipid isolated from the marine sponge Agelas mauritianus, is a potent immunomodulator with therapeutic potential for the treatment of autoimmune diseases and cancer. The Toll-like receptor 4 (TLR4), one of the promising molecular targets for immune-modulating drugs, is commonly expressed in innate immune cells especially macrophages and dendritic cells. Currently, whether alpha-GalCer can activate TLR4 signaling pathways remains unreported. In this study, we examined the effects of alpha-GalCer and its various structural analogs, CCL-1 approximately 47, on TLR4 activation. We found that one alpha-GalCer analog (CCL-34), but not alpha-GalCer itself, strongly stimulated NF-kappaB activity in RAW 264.7 cells. CCL-34 activated NF-kappaB in a TLR4-dependent manner and stimulated TNF-alpha production in bone marrow cells of TLR4-functional C3H/HeN mice but not in those of TLR4-defective C3H/HeJ mice. Furthermore, CCL-34 treatment stimulated NF-kappaB activation and IL-8 production in a 293 cell line constitutively expressing human TLR4, MD-2 and CD14. Treatment of RAW 264.7 cells with CCL-34 also activated TLR4-downstream mitogen-activated protein kinases (ERK, JNK and p38), induced expression of TLR4-downstream genes (TNF-alpha, IL-6, IL-1beta and iNOS) and promoted production of cytokines characteristic of activated macrophages. CCL-34-treated RAW 264.7 cells acquired a distinct morphology similar to that of LPS-activated macrophages and exhibited higher phagocytotic activity. Moreover, treatment with a TLR4-neutalizing antibody inhibited the CCL-34-induced morphological alteration. In summary, we identify a novel synthetic compound CCL-34 that can activate macrophages via TLR4-dependent signaling pathways. Our results suggest that CCL-34 is an immune modulator and may serve as a potential drug lead for immunotherapy.     

肺癌患者GMCSF检测及临床意义

应用北京华英放免试剂盒检测 78例肺癌患者血清、胸水中粒细胞 巨噬细胞集落刺激因子 (ＧＭ ＣＳＦ )浓度 ,旨在探讨不同类型肺癌及病情程度与ＧＭ ＣＳＦ的关系。结果见表 1 ,2。　　ＧＭ ＣＳＦ细胞因子目前已广泛应用于感染、肿瘤化疗后粒细胞减少症等的治疗 ,而有关ＧＭ ＣＳＦ检测在肺癌不同病期的结果 ,尚未见报道。本实验结果说明 ,肺癌患者ＧＭ ＣＳＦ明显低于正常对照组 ,而不同类型肺癌之间无明显差异 ,但从表 2可以看出 ,随着肺癌患者的临床症状不断恶化ＧＭ ＣＳＦ结果也逐渐降低。鉴于以上情况 ,适当应用ＧＭ ＣＳＦ可能有助…     

γ射线诱导癌细胞凋亡致敏树突状细胞后的免疫应答

目的 观察树突状细胞（DCs）从γ射线诱导的凋亡胆管癌细胞获取抗原后，抗肿瘤免疫应答及对胆管癌细胞的特异性免疫杀伤效果。方法 用粒－巨噬细胞集落刺激因子（GM－CSF）加白介素－4（IL－4）从人外周血分化、诱导DCs,γ射线在体外诱导培养的人胆管癌细胞凋亡，将DCs、T淋巴细胞和凋亡胆管癌细胞共培养，同时设计不同类型肿瘤细胞（坏死胆管癌细胞及培养胆管癌细胞）作对照，7d后，分离、富集DCs、T淋巴细胞进行免疫应答及肿瘤细胞杀伤试验。结果 与凋亡胆管癌细胞共培养之DCs可以有效提呈胆管癌细胞抗原，有强烈的免疫应答，刺激的细胞毒T淋巴细胞（CTLs)特异性地杀伤胆管癌细胞。结论 γ射线诱导癌细胞凋亡可以致敏rhGM-CSF加rhIL-4从人外周血单核细胞诱导、扩增出的DCs并产生显著的杀伤胆管癌细胞的免疫反应，可望成为特异性免疫治疗肿瘤的一条新途径。     

含凝血酶识别位点的重组人粒细胞/巨噬细胞集落刺激因子在大肠杆菌的表达

采用PCR技术，从细胞中获取粒细胞／巨噬细胞集落刺激因子cDNA，并在5′端设计上凝血酸酶切位点、5个组氨酸密码子、起始密码子及EcoR1酶切位点，在3′端设计上BamH1酶切位点。将扩增产物酶切后克隆到pBV220质粒的EcoRⅠ－BamHⅠ酶切位点，转化大肠杆菌DH5α，筛选阳性重组子（pGM09／DH5α）。在E.coli中获高效表达，表达量占总蛋白的31．2％，生物活性为1．1×107U／L发酵液，纯化工艺简化。     

Granulocyte–macrophage colony-stimulating factor promotes survival of dopaminergic neurons in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced murine Parkinson's disease model

Granulocyte–macrophage colony-stimulating factor (GM-CSF) is a hematopoietic cytokine that has the potential for clinical application. The biological effects of GM-CSF have been well characterized, and include stimulation of bone marrow hematopoietic stem cell proliferation and inhibition of apoptosis of hematopoietic cells. In contrast, the therapeutic effects of GM-CSF on the central nervous system in acute injury such as stroke and spinal cord injury have been reported only recently. To better understand the protective effect of GM-CSF on dopaminergic neurons in Parkinson's disease (PD), we investigated the effect of GM-CSF on the survival of dopamine neurons and changes in locomotor behavior in a murine PD model. We investigated the neuroprotective effects of GM-CSF in 1-methyl-4-phenylpyridinium (MPP⁺)-treated PC12 cells as well as in embryonic mouse primary mesencephalic neurons (PMNs) in vitro . To investigate the role of GM-CSF in vivo , we prepared a mouse 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) PD model, and examined the effects of GM-CSF on dopaminergic neuron survival in the substantia nigra and on locomotor behavior. Treatment with GM-CSF significantly reduced MPP⁺-induced dopaminergic cell death in PC12 cells and PMNs in vitro . GM-CSF modulated the expression of apoptosis-related proteins, Bcl-2 and Bax, in vitro . Furthermore, administration of GM-CSF (50 μg/kg body weight/day) in vivo for 7 days protected dopaminergic neurons in the substantia nigra and improved locomotor behavior in a mouse MPTP model of PD.     

Mixed haematopoietic colony formation via immature blast cell clusters on foetal mesenchymal cell layers distinguishes stem cells from peripheral blood,cord blood,bone marrow and blood stem cells mobilized by granulocyte-macrophage colony-stimulating factor

Abstract: Multilineage colony formation was evaluated from healthy donors' bone marrow (BM), peripheral blood (PB) and cord blood (CB) and compared with blood stem cell (BSC) harvests of sarcoma patients mobilized with granulocyte-macrophage colony-stimulating factor (GM-CSF). The test was a modified CFU-blast assay performed with and without an irradiated foetal mesenchymal cell layer (HFFF). These non-transformed mesenchymal cells served as a good source of haematopoietically active stroma cells in that cytokine expression patterns (interleukin (IL)-6, granulocyte (G)-CSF, GM-CSF) and adhesion molecules on HFFF cells were qualitatively identical to BM-derived fibroblasts, but the expression density of adhesion receptors was significantly higher. This HFFF layer stimulated blood stem cells of GM-CSF-treated patients significantly more than a cocktail of exogenous growth factors with IL-1, IL-6, and stem cell factor (SCF). The reverse was true for multilineage colonies from healthy donors' PB, BM, and CB. According to these results, stem cells of GM-CSF-treated patients are functionally distinct due to their dependence on stroma-derived factors and/or matrix-adhesion interactions and can be reproducibly evaluated on these mesenchymal cells.     

Recombinant human granulocyte macrophage colony stimulating factor following alternating non cross resistant chemotherapy in Hodgkin's disease.

Fourteen patients with Hodgkin's disease (two previously untreated, 12 following relapse or with refractory disease) were treated with a combination chemotherapy regimen comprising chlorambucil, vinblastine, procarbazine, prednisolone, etoposide, vincristine and adriamycin administered on days 1-8. Recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) (mammalian glycosylated, Sandoz/Schering-Plough) was administered after alternate cycles of chemotherapy from day 10 for 7 days by continuous intravenous (i.v.) infusion in 12 patients in a dose finding study (dose: 2 micrograms/kg/day in four patients, 4 micrograms/kg/day in four patients and 8 micrograms/kg/day in four patients) and by daily subcutaneous (s/c) injections in two patients (8 micrograms/kg/day). There was a rapid peripheral leucocytosis following the rhGM-CSF, reaching a peak at 1-2 days in 12/14 patients. The initial leucocytosis was composed of neutrophils followed by a rise in immature myeloid cells. There was no difference observed in the duration or depth of the nadir following chemotherapy or in the rate of recovery of peripheral white cell counts between cycles with and without rhGM-CSF in patients treated with 2 and 4 micrograms/kg/day. At the dose of 8 micrograms/kg/day, 3/6 patients had a shorter nadir duration in the cycle with rhGM-CSF, compared with cycle without rhGM-CSF. There was no difference in frequency of infection in cycles with and without rhGM-CSF. Following chemotherapy, six patients achieved clinical remission, six partial remission and two had progressive disease.