Flt3-L是否为体外制备急性髓系白血病患者DC瘤苗的更佳选择?

目的:通过对体外使用Flt3-L(FL)及粒-巨噬细胞集落刺激因子(GM-CSF)诱导完全缓解期急性髓系白血病(AML-CR)患者的骨髓中单个核细胞(BMNC)分化为树突状细胞(DC)、分泌白介素(IL-12P70),干扰素-α(INF-α)的情况,及所诱导DC体外刺激自体T细胞(auto-T cells)活化增殖能力的观察,进一步了解FL在DC瘤苗制备中的作用特点。方法:分离AML-CR患者BMNC采用FL／GM-CSF单因子培养于RPMI1640完全培养基中至第8天,加入LPS刺激活化后流式细胞仪明确DC免疫表型,ELISA法测定培养液上清中IL-12P70、INF-α,收集阳性细胞体外激发增埴auto-T细胞,MTT法测试T细胞增殖情况。结果:10例AML患者的BMNC经FL／GM-CSF分别培养至第72 h均开始有细胞成簇生长,流式细胞检测显示CD1a、CD83、CD80、CD86、HLA-DR均表达明显上调,ELISA测定FL组IL-12P70,INF-α分泌量较GM-CSF组有显著增加。2组均可产生较强的T细胞增殖效应,与GM-CSF比,FL组对CD4 T细胞的增殖作用更强。结论:在体外,FL可更强诱导AML-CR患者BMNC产生DC,同时分泌大量IL-12P70及INF-α。故与GM-CSF相比,FL可能具有更佳的抗肿瘤免疫作用。     

Regulation of immunomodulatory functions by granulocyte-macrophage colony-stimulating factor and granulocyte colony-stimulating factor in vivo

 The present study was designed to investigate in vivo immunomodulatory properties of hematopoietic growth factors. The influence on the activation of cytokine synthesis and on the expression of surface antigens associated with cellular activation of G-CSF or GM-CSF was investigated in cancer patients receiving these factors. One single dose of growth factor was administered to patients with bladder cancer (G-CSF group) or small cell lung cancer (GM-CSF group) before chemotherapy. After cytoreductive chemotherapy patients received supportive therapy with G-CSF or GM-CSF. Peripheral blood mononuclear cells and plasma samples were obtained for flow cytometry, Northern blot analysis, and assessment of cytokine protein levels after single-dose as well as after continous cytokine administration. Our results demonstrate differences in the induction of biological activities by GM-CSF and G-CSF in vivo which correlate well with in vitro findings. Among mature hematopoietic cells the effect of G-CSF is restricted to the granulocyte lineage. With GM-CSF moderate but unequivocal modulation of monocyte function was observed. On peripheral blood monocytes expression of MHC class-II molecules and CD44 was markedly stimulated. After one single dose of GM-CSF, plasma levels of sCD25 and IL-1RA were significantly induced (p<0.0001, p=0.032, respectively) and a trend to increased IL-8 levels was observed. The changes in plasma proteins were not correlated with shifts of mRNA expression for IL-8 and IL-1RA. T-cell activation was not observed with either cytokine. These results suggest that immunomodulatory features are differentially regulated by G-CSF and GM-CSF. The clinical relevance of a selective use of both hematopoietic growth factors in various disease settings remains to be determined. Received: 20 March 1996 / Accepted: 19 July 1996     

Effect of granulocyte-macrophage colony-stimulating factor on hepatic regeneration after 70% hepatectomy in normal and cirrhotic rats

BackgroundPost-hepatectomy liver insufficiency is one of the most serious postoperative problems and its prevention is important after major hepatic resection, especially in the cirrhotic liver. Some growth factors and cytokines appear to play important roles in liver regeneration. In the present study we have investigated the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on hepatic regeneration after 70% partial hepatectomy (PH) in cirrhotic and non-cirrhotic rats.MethodsA rat model of liver cirrhosis was prepared using thioacetamide (TAA) (a dose of 20 mg/100 g body w, intra-peritoneally) on three days a week for 12 weeks. Adult male rats were divided into four groups:Group 1 (n=10) no cirrhosis and no GM-CSF; Group 2 (n=10) no cirrhosis and GM-CSF; Group 3 (n=10) cirrhosis and no GM-CSF; and Group 4 (n=10) cirrhosis and GM-CSF. All the rats underwent a 70% hepatectomy, and GM-CSF was administrated immediately after operation in Groups 2 and 4. On postoperative days 2 and 7, fresh samples from the remnant liver were obtained to evaluate its regenerative capacity.The liver regenerative process was estimated by DNA synthesis, using flow cytometry.ResultsProliferation index (PI) of hepatocytes at 48 h was higher in Group 4 rats than Group 3 rats (p<0.05). On postoperative day 7, PI was elevated in Group 3 rats compared with Group 4 rats, but this difference was not statistically significant. In non-cirrhotic rats given GM-CSF, PI was increased compared with Group 1 rats at day 2 (p<0.05), but not at day 7.ConclusionsThe findings suggest that the proliferative capacity of liver cells is impaired and delayed after 70% PH in cirrhotic rat liver. GM-CSF administration might enhance the liver PI in both normal and TAA-induced cirrhotic rats.     

CD34+ peripheral blood progenitor cell and monocyte derived dendritic cells: a comparative analysis

Dendritic cells (DC) have been generated in vitro from either CD34⁺ haemopoietic progenitor cells (HPC) or peripheral blood monocytes (Mo) in the presence of specific cytokine combinations, including granulocyte-macrophage colony-stimulating factor (GM-CSF). Since differences between DC from either source may be important for the clinical use of these antigen-presenting cells (APC), a comparative analysis was performed. HPC were expanded in the presence of interleukin (IL)-3, IL-6 and stem cell factor (SCF) (days 1–7) and subsequently induced by IL-4 + GM-CSF (days 8–26) to differentiate to Langerhans-type cells (pLC). The latter cytokines were similarly used to generate Mo-derived LC (mLC). Maturation of both cell types, pLC and mLC, to interdigitating DC-type cells (iDC) was induced by tumour necrosis factor-α (TNF-α) or lipopolysaccharide (LPS). Analysis of mLC/pLC and miDC/piDC with respect to morphology, phenotype, antigen uptake and presentation revealed a high similarity of DC from either source. The majority of mLC, however, exhibited a more mature differentiation stage, compared to pLC, evidenced from lower numbers of multilaminar MHC class II compartments and less efficient APC function for extracellular protein antigens. Although macropinocytosis was performed by LC, neither LC nor iDC from either source were able to take up 0.5 μm latex beads. However, phagocytosis of 0.5 μm and 1 μm beads was performed by Mo that could subsequently be induced to become iDC, thus providing the unique opportunity to present phagocytosed material in DC-type fashion. Mo may be the preferential source for clinical use of iDC-type cells since preparation and culture are easier to perform and are less costly while APC function is similar to HPC-derived iDC.     

GM-CSF及其受体与早期自然流产的关系研究

目的：探讨母胎界面局部细胞因子GM-CSF及其受体与早期自然流产的发生的关系．方法采集2009年8月至2011年12月间来云南省第三人民医院进行人工流产及自然流产病例的绒毛及脱膜组织各30例,用放射免疫法检测2组外周血中绒毛膜促性腺激素（HCG）含量,利用免疫组织化学和Western blot检测绒毛及脱膜组织中巨噬细胞集落刺激因子（GM-CSF）及其受体（GM-CSFR）表达情况．结果自然流产组HCG值与人工流产组相比明显下降（P〈0.05）．绒毛及脱膜组织中GM-CSF和GM-CSFR 2组均有表达．与人工流产组相比,绒毛组织中,自然流产组的 GM-CSFR增加（P〈0.05）,GM-CSF的表达上调,但无统计学意义（＞0.05）．脱膜组织中,自然流产组的GM-CSF和GM-CSFR均增加（P〈0.05）．结论在脱膜组织中,适量的GM-CSF和GM-CSFR浓度维持妊娠的继续,浓度过高则不利于妊娠,可能是自然流产的原因之一．     

Recombinant human granulocyte-macrophage colony-stimulating factor hydrogel promotes healing of deep partial thickness burn wounds

Objective

To assess the effects of recombinant human granulocyte/macrophage colony-stimulating factor (rhGM-CSF) hydrogel on the healing of deep partial thickness burn wounds.

Methods

Ninety three wounds of 65 burn patients who suffered from a deep partial thickness burn of <5% TBSA and could not heal over 3 weeks were included in this study. The patients were randomly assigned to use rhGM-CSF hydrogel (GC group, n = 32) or hydrogel without rhGM-CSF (control group, n = 33). rhGM-CSF hydrogel or hydrogel without rhGM-CSF was topically applied to the wounds, the dressing was changed once a day. Wound healing time and percentage, wound discharge, periwound inflammation, the positive wound swabs culture count, and adverse drug reactions were observed and compared between two groups.

Results

Healing time was 12.2 ± 5.0 days after the application of rhGM-CSF hydrogel. This was significantly shorter than that of control wounds (15.5 ± 4.7 days). Healing percentage at 14 days in the rhGM-CSF-treated wounds was 97.5 ± 7.7%, which was markedly higher than the control (85.9 ± 6.8%). At 3, 6, 12, 14 day, the GC group was significantly superior to the control group with respect to the score of periwound inflammation, wound purulence and discharge. The positive wound swabs culture count of the GC group on the 7th and 14th day post-treatment was 14 and 4, respectively, which was significantly lower than the control.

Conclusion

rhGM-CSF hydrogel promotes the healing process of deep partial thickness burns effectively. No adverse reaction of the drug was observed during the study.     

氧化／抗氧化失衡在COPD稳定期慢性肺损伤的机制研究

目的探讨稳定期不同级别COPD患者血清中氧化／抗氧化因子及细胞炎症因子水平与肺功能主要指标的关系。方法选取COPD稳定期患者80例,检测各患者肺功能与血清中反应性氧核素（ROS）、超氧化物歧化酶（SOD）及细胞炎症因子水平,观测各组的差异并分析其相关性。结果FEV1、FVC、FEV1／FVC、SOD、ROS、TNF．d、IL-8、GM—CSF水平在不同组别中存在差异（P〈0．05）,ROS、TNF-a、IL-8、GM—CSF水平与FEV1,FVC、FEV1／FVC存在着明显的负相关（P〈0．05）,SOD与其存在明显正相关（P〈0．05）。结论COPD细胞炎症因子的持续升高及氧化加重可能是引起稳定期COPD肺组织慢性损伤、功能降低的重要机制。     

GM-CSF in the treatment of Felty syndrome

Many therapeutic agents have been tried with variable success in the treatment of Felty neutropenia, but the reports are anecdotal. We now describe the second trial of recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF), in a splenectomized, infected patient with Felty syndrome.     

GM-CSF对膀胱癌EJ细胞株CD44v6表达的影响

目的：明确GM－CSF对EJ细胞增殖及对CD44v6表达的影响。方法：MTT法检测GM－CSF和IFN对EJ细胞增殖的影响，免疫细胞化学法观察GM－CSF对CD44v6表达的影响。结果：GM－CSF以不同浓度作用EJ细胞对其增殖无影响，但GM－CSF联用IFN可抑制J细胞增殖，GM－CSF连用IF可降低CD44v6表达。结果：GM－CSF＋IFN可降低CD44v6在EJ细胞中的表达。     

Effects of granulocyte-macrophage colony stimulating factor produced in Chinese hamster ovary cells (regramostim), Escherichia coli (molgramostim) and yeast (sargramostim) on priming peripheral blood progenitor cells for use with autologous Done marrow after high-dose chemotherapy

Abstract: Peripheral blood progenitor cells (PBPCs) were collected without prior association with chemotherapy but after the administration of granulocyte-macrophage colony-stimulating factor (GM-CSF) produced in Chinese hamster ovary cells (CHO-GM, regramostim), Escherichia coli (E. coli-GM, molgramostim), or yeast (Yeast-GM, sargramostim) and used in conjunction with autologous bone marrow after high-dose chemotherapy in 69 patients with breast cancer or melanoma. The mean peripheral white blood cell (WBC) counts increased by 2.2 to 2.7-fold after regramostim, 4.5 to 7.3-fold after molgramostim and 4.3-fold after sargramostim. All patients underwent three leukaphereses. The mean (& standard error) total nucleated pheresed cells per kg × 10⁸ were 4.15 & 0.56, 15.10 & 1.77 and 7.24 & 1.00 for patients receiving regramostim, molgramostim or sargramostim respectively. The mean (& standard error) granulocyte-macrophage colony-forming units per kg × 10⁴ mobilized into the PB were 8.75 & 3.63, 71.03 & 17.85, and 65.11 & 18.74 for patients receiving regramostim, molgramostim, or sargramostim respectively. The total mean (& standard error) CD34+ cells per kg × 10⁷ collected by three leukaphereses were 3.28 & 1.62, 1.34 & 0.51 and 2.57 & 1.93, for patients receiving regramostim, molgramostim or sargramostim respectively. The use of either molgramostim- or sargramostim-primed PBPCs led to complete elimination of absolute leukopenia with a WBC count under 100/mm³ in 64% and 77% of patients treated, respectively. Patients receiving molgramostim-primed PBPCs required fewer red blood cells transfusions than patients receiving regramostim-primed PBPCs (p = 0.0062). Our data indicate that PBPCs collected without prior association with chemotherapy but after either molgramostim or sargramostim with autologous bone marrow support and GM-CSF shorten the hematopoietic recovery after myeloablative chemotherapy in patients with breast cancer or melanoma.