粒细胞-巨噬细胞集落刺激因子基因修饰的树突状细胞疫苗增强体外抗肿瘤免疫效应

Objective: The aim of the study was to investigate whether dendritic cell (DC) precursors, recruited by injection of chemokine ligand 3 (CCL3), induce enhanced anti-tumor immunity after granulocyte-macrophage colony stimulating factor (GM-CSF) transfection in mice ex vivo. Methods: The 615 mice were injected with CCL3 via the tail vein. Freshly isolated B220–CD11c+ cells were cultured with cytokines. For adenoviral (Ad)-mediated gene transduction, DCs were transferred AdGM-CSF gene at different ratios of mu...     

粒细胞-巨噬细胞集落刺激因子基因修饰的树突状细胞疫苗增强体外抗肿瘤免疫效应

目的 研究粒细胞-巨噬细胞集落刺激因子(GM-CSF)基因修饰树突状细胞(DC)后形态、表型及功能的变化,以及增强DC疫苗对肿瘤细胞的体外杀伤作用.方法 小鼠尾静脉注射趋化因子配体3(CCL3),分选得到B220- CDllc+细胞,经细胞因子培养诱导分化DC.在体外用含GM-CSF基因的重组腺病毒(AdGM-CSF)转染DC,酶联免疫吸附试验(ELJSA)检测转染后GM-CSF的水平.通过细胞形态学观察、表型分析及混合淋巴细胞反应(MLR),检测GM-CSF基因修饰前后DC的变化.反复冻融法制备胃癌可溶性抗原,将其与GM-CSF基因修饰的DC共同培养,制备DC疫苗,四甲基偶氮唑蓝(MTT)法检测活化的T淋巴细胞在体外对小鼠前胃癌细胞(MFC)的杀伤作用,ELISA法检测干扰素γ(INF-γ)的分泌情况.结果 CCL3注射后,外周血中B220- CD11c+细胞明显增加,48 h达到高峰[占外周血单个核细胞的(13.88±1.10)%].AdGM-CSF转染后,培养液上清中GM-CSF浓度升高,当感染复数(MOI)为1:100时达到高峰[(130.00±12.61)pg/m1].经GM-CSF.基因修饰的DC在形态上更趋成熟,MHCⅡ类分子、CD80、CD86等细胞表型明显上调,具有更强的刺激T细胞增殖的能力.荷载胃癌抗原的DC激活的T淋巴细胞对MFC细胞具有特异性杀伤作用,并产生高水平的INF-γ[(1245.00±13.75)pg/ml].结论 GM-CSF转染DC后,能大量表达GM-CSF,DC形态及细胞表型更趋成熟,刺激T细胞增殖能力明显增强.GM-CSF基因修饰的DC在体外可诱导出针对靶肿瘤细胞的特异性杀伤作用.     

粒细胞-巨噬细胞集落刺激因子基因修饰的树突状细胞疫苗增强体外抗肿瘤免疫效应

Objective To investigate if granulocyte-macrophage colony stimulating factor (GM-CSF) gene-modified dendritic cells ( DC) enhance antitumor immunity in vitro. Methods Mice were injected with chemokine ligand 3 (CCL3) via the tail vein. Fresh B220-CD11c+ cells were sorted from the peripheral blood mononuclear cells (PBMCs) and cultured into DCs by cytokines. DCs were transfected with AdGM-CSF gene at different ratios of multiplicity of infection ( MOI) to determine the optimal gene transfection conditions, and the expression of GM-CSF was detected after transfection. The variation of GM-CSF gene-modifiedDCs were analyzed by morphological examination, phenotype analysis, and mixed lymphocyte reaction (MLR). DCs were loaded with gastric cancer antigen obtained by freezing and thawing method. The killing effect of DCs vaccine-stimulated T lymphocytes on gastric cancer cells was assessed by MTT assay. INF-γ production was determined with the INF-γ ELISA kit. Results B220- CD11c+ cells increased obviously after CCL3 injection. The ELISA results showed that after GM-CSF gene modification, DCs could produce high level of GM-CSF. When DCs were transfected with AdGM-CSF gene at MOI equal to 100, the GM-CSF level in culture supematants reached saturation [(130.00±12.61) pg/ml]. After GM-CSF gene-modification, DCs tend to be more maturated as detected by morphological observation and phenotype analysis. At the same time, the capacity of activating the proliferation of allogeneic T lymphocytes was enhanced greatly. T lymphocytes stimulated by DCs transfected with GM-CSF gene showed a specific killing effect on gastric carcinoma cells and produced high level of INF-γ[ ( 1245. 00±13. 75) pg/ml].Conclusion After GM-CSF gene modification, DCs can produce high level of GM-CSF, which tend to be more maturated, and the capacity of activating the proliferation of allogeneic T lymphocytes is enhanced greatly. GM-CSF gene modified DCs can induce specific CTL to target tumor cells in vitro.     

粒细胞-巨噬细胞集落刺激因子基因修饰的树突状细胞疫苗增强体外抗肿瘤免疫效应

Objective To investigate if granulocyte-macrophage colony stimulating factor (GM-CSF) gene-modified dendritic cells ( DC) enhance antitumor immunity in vitro. Methods Mice were injected with chemokine ligand 3 (CCL3) via the tail vein. Fresh B220-CD11c+ cells were sorted from the peripheral blood mononuclear cells (PBMCs) and cultured into DCs by cytokines. DCs were transfected with AdGM-CSF gene at different ratios of multiplicity of infection ( MOI) to determine the optimal gene transfection conditions, and the expression of GM-CSF was detected after transfection. The variation of GM-CSF gene-modifiedDCs were analyzed by morphological examination, phenotype analysis, and mixed lymphocyte reaction (MLR). DCs were loaded with gastric cancer antigen obtained by freezing and thawing method. The killing effect of DCs vaccine-stimulated T lymphocytes on gastric cancer cells was assessed by MTT assay. INF-γ production was determined with the INF-γ ELISA kit. Results B220- CD11c+ cells increased obviously after CCL3 injection. The ELISA results showed that after GM-CSF gene modification, DCs could produce high level of GM-CSF. When DCs were transfected with AdGM-CSF gene at MOI equal to 100, the GM-CSF level in culture supematants reached saturation [(130.00±12.61) pg/ml]. After GM-CSF gene-modification, DCs tend to be more maturated as detected by morphological observation and phenotype analysis. At the same time, the capacity of activating the proliferation of allogeneic T lymphocytes was enhanced greatly. T lymphocytes stimulated by DCs transfected with GM-CSF gene showed a specific killing effect on gastric carcinoma cells and produced high level of INF-γ[ ( 1245. 00±13. 75) pg/ml].Conclusion After GM-CSF gene modification, DCs can produce high level of GM-CSF, which tend to be more maturated, and the capacity of activating the proliferation of allogeneic T lymphocytes is enhanced greatly. GM-CSF gene modified DCs can induce specific CTL to target tumor cells in vitro.     

集落刺激因子对白血病细胞的作用

集落刺激因子(Colony Stimulating Factor,CSF)又称生长因子(Growth Factor,GF)是体内生物性造血生长因子,在调节髓系细胞的增殖和分化过程中发挥着重要作用。现已发现CSF还具有     

重组人粒系集落刺激因子对白血病细胞的作用

作者观察了重组人粒系集落刺激因子（ｒｈＧ－ＣＳＦ）对ＨＬ－６０细胞和原代培养的急性髓系白血病（ＡＭＬ）细胞的作用，结果显示，ｒｈＧ－ＣＳＦ可刺激白血病细胞生长，并诱导部分ＡＭＬ细胞向粒系分化。提示ｒｈＧ－ＣＳＦ对白血病细胞存在着增殖与分化的双重作用，这种作用对改进白血病治疗可能产生重要影响。     

低剂量粒细胞集落刺激因子动员健康供者造血干细胞的研究

 目的　探讨低剂量（5 μg·kg-1·d-1）粒细胞集落刺激因子（G-CSF）对健康供者CD+34细胞动员的效果及造血干细胞最佳采集时间。方法　对2006年2月至2009年4月108例健康供者给予G-CSF 5 μg·kg-1·d-1,在动员后的第4天至第6天收集标本,检测相应时间点的外周血白细胞（WBC）计数、采集物单个核细胞（MNC）计数、采集物中CD+34细胞的比例、粒-巨噬细胞集落形成单位（CFU-GM）培养。结果　动员后采集物中第4天至第6天CD+34细胞比例分别为（0.71±0.08）%、（1.09±0.09）%、（0.57±0.08）%,第4天至第6天CFU-GM产率分别为：（93.33±44.51）／105 MNC、（124.61±57.85）／105 MNC和（80.25±49.24）／105 MNC。CD+34细胞比例和CFU-GM产率均在第5天达到峰值（P＜0.05）,第4天次之（P＜0.05）,第6天再次之（P＜0.05）。动员后采集物中第4天至第6天CD+34细胞量分别为（3.33±1.36）×106／kg、（4.14±1.67）×106／kg、（2.79±1.47）×106／kg,第5天达到峰值（P＜0.05）,第4天次之（P＜0.05）,第6天再次之（P＜0.05）。108例供者采集2次（第4天、第5天）均可满足移植标准。所有供者均无严重不良反应发生。结论　单一低剂量G-CSF能够有效动员健康供者造血干细胞。动员后于第4、第5天行干细胞采集优于第5、第6天。     

粒细胞-巨噬细胞集落刺激因子联合放疗研究进展

粒细胞-巨噬细胞集落刺激因子（granulocyte-macrophage colony-stimulating factor,GM-CSF）作为一种造血因子可以有效诱导多种具有抑瘤效应的免疫细胞增殖,从而发挥抗肿瘤免疫反应。放疗作为肿瘤治疗的主要手段之一,不仅可以直接杀伤肿瘤细胞,而且会对抗肿瘤免疫产生影响。多项临床研究提示,放疗联合GM-CSF可以诱导产生旁观者效应并增强对肿瘤的远期控制,从而增强放疗的抗肿瘤效应。本文就GM-CSF及GM-CSF联合放疗的研究进展予以综述。      

粒细胞集落刺激因子影响肿瘤转移的研究进展

粒细胞集落刺激因子（granulocyte colony stimulating factor,G-CSF）也称为集落刺激因子3（colony stimulating factor 3,CSF3）,是一种含有174个氨基酸的糖蛋白（19.6 KDa）。G-CSF是控制骨髓祖细胞增殖和分化为中性粒细胞的主要造血细胞因子。当宿主受到感染或有组织损伤时,G-CSF可动员中性粒细胞。临床上G-CSF多用于放、化疗患者,预防或治疗放化疗后所造成的中性粒细胞减少症或发热性中性粒细胞减少症。G-CSF可通过动员肿瘤相关中性粒细胞促进肿瘤生长和转移。本文就G-CSF对肿瘤生长、转移的促进作用和可能机制进行综述。      

重组人集落刺激因子对脐带血和骨髓造血祖细胞体外培养的影响

分别应用单层琼脂培养中甲基纤维素培养方法，同期观察了重组人粒细胞集落刺激因子，重组人粒单细胞集落刺激因子，重组人白细胞介素和红细胞生成素，对脐带血和骨髓造血祖细胞的影响。结果显示：脐带血中含有丰富的造血祖细胞，其早期的造血祖细胞数量与骨髓相似，脐带血的造血祖细胞的增殖能力更强。     

粒细胞集落刺激因子的临床应用

目前在国内市场上销售的粒细胞集落刺激因子(G-CSF)出自三家公司(见表1)。 从表1可以看出同样是G-CSF,商品名称不同,结构上也稍有差异,但具有同等的生物学活性,即能刺激骨髓组织产生嗜中性粒细胞,可有效地治疗一切伴有造血细胞障碍或衰竭的疾病。是肿瘤辅助治疗的有效药物之一。目前已在临床上广泛应用。     

应用重组人粒细胞集落刺激因子的经验与教训专题讨论

重组人粒细胞集落刺激因子（rhG-CSF）在我国已是治疗血液系统疾病非常重要的手段之一;近些年,其对非血液系统疾病的治疗也取得了很大进展。rhG-CSF的应用在临床已积累了许多丰富的经验,收到了是好的疗效。但个别医生,由于对G-CSF缺乏全面了解和认识,招致一些原可以避免的严重并发症,如诱发肺、脑、心肌梗死或促进和诱发粒细胞白血病等。如何把握G—CSF应用的指征,扬其优越效果,避其毒副作用,确保医疗安全,是值得我们深入探讨的问题,也是一个现实而有意义的工作。 鉴于此,我刊在本期约请了田丁教授和其他5位专家,对上述问题进行了专题讨论。作者们结合自己丰富的临床经验和渊博的学识,除了介绍G—CSF治疗疾病的成功经验外,着重提出一些如何避免或减少并发症的见解。所有这些,对血液学同道特别对年轻的医生们,无疑是值得借鉴的。[编者按]     

重组人集落刺激因子对脐带血和骨髓造血祖细胞体外培养的影响

分别应用单层琼脂培养和甲基纤维素培养方法，同期观察了重组人粒细胞集落刺激因子（ｒｈＧ－ＣＳＦ）、重组人粒单细胞集落刺激因子（ｒｈＧ－ＣＳＦ）、重组人白细胞介素－３（ｒｈＩＬ－３）和红细胞生成素（ＥＰＯ），对脐带血和骨髓造血祖细胞的影响。结果显示：脐带血中含有丰富的造血祖细胞，其早期的造血祖细胞数量与骨髓相似，脐带血的造血祖细胞的增殖能力更强。     

粒细胞集落刺激因子动员外周血造血干细胞致急性肺损伤1例

患者,女,32岁,确诊非霍奇金淋巴瘤Ⅳ期(非特异性外周T细胞型)2个月,于2002年12月7日入院行自体外周血造血干细胞移植。既往体健。入院后查血常规、胸片、心电图、肺功能等均正常。遂于12月10日开始外周血干细胞动员,方案为NAP(米托蒽醌 阿糖胞苷 泼尼松)化疗及G-CSF(粒细胞集落刺激因子)联合,G-CSF自化疗结束后24h始用,     

集落刺激因子在小细胞肺癌化疗中的辅助作用

骨髓抑制是小细胞肺癌（ＳＣＬＣ）化疗的主要毒副作用。集落刺激因子（ＣＳＦ）对促进中性粒细胞恢复、防治粒细胞缺乏症和增加剂量敏感性都具有良好作用。本文综述ＣＳＦ治疗ＳＣＬＣ的理论基础和临床应用新进展。     

粒细胞集落刺激因子治愈化学性皮肤坏死1例

化疗药物渗漏到皮下组织,轻者可引起局部红肿、疼痛,严重可损伤神经、肌腱,甚至造成皮肤组织坏死、肢体功能障碍。我们采用以粒细胞集落刺激因子（G-CSF）为主的综合治疗治愈表阿霉素外渗导致皮肤坏死1例,现报告如下。     

粒细胞集落刺激因子的免疫调节作用研究进展

粒细胞集落刺激因子（G-CSF）主要用于各种原因引起的粒细胞减少症及外周血干细胞的动员。近年来关于G-CSF免疫调节作用及应用成为了研究热点。全文综述了G-CSF对T细胞、树突状细胞、Treg、NK细胞、细胞因子等的调节作用及其在免疫相关性疾病治疗中的应用。     

粒细胞巨噬细胞集落刺激因子作为肿瘤疫苗佐剂的研究进展

研制肿瘤疫苗通过激活患者自身免疫系统以清除或控制肿瘤是当前的研究热点。然而肿瘤疫苗研制中存在的一个主要问题是大多数肿瘤的免疫原性很弱,不能正常地呈递抗原以激活细胞毒性T淋细胞（CTL）,为了提高其免疫原性,常需加入佐剂。大量研究表明,粒细胞巨噬细胞集落刺激因子（GM-CSF）以其显著的免疫调节作用和低毒性成为最具吸引力的免疫佐剂。     

粒细胞巨噬细胞集落刺激因子作为肿瘤疫苗佐剂的研究进展

研制肿瘤疫苗通过激活患者自身免疫系统以清除或控制肿瘤是当前的研究热点。然而肿瘤疫苗研制中存在的一个主要问题是大多数肿瘤的免疫原性很弱,不能正常地呈递抗原以激活细胞毒性T淋细胞(CTL),为了提高其免疫原性,常需加入佐剂。大量研究表明,粒细胞巨噬细胞集落刺激因子(GM-CSF)以其显著的免疫调节作用和低毒性成为最具吸引力的免疫佐剂。