粒细胞-巨噬细胞集落刺激因子在肺部疾病的研究进展

粒细胞-巨噬细胞集落刺激因子（granulocytemacrophage colony-stimulating factor, GM-CSF）是一种多功能造血因子,属于糖蛋白因子家族。主要由活化的T细胞、树突状细胞、巨噬细胞、角质细胞、内皮细胞和成纤维细胞等分泌,并通过自分泌和旁分泌方式作用于这些细胞,从而刺激粒细胞和巨噬细胞增殖、成熟和分化,具有极重要的免疫调节功能。现就GM—CSF的生理、生化特性及它在肺部疾病中的研究进展作阐述。     

重组人粒细胞巨噬细胞集落刺激因子临床应用现况及前景

重组人粒细胞巨噬细胞集落刺激因子临床应用现况及前景林榕，楼方定ＪｏｎｅｓＴ．Ｃ１９９３年９月９～１１日在瑞士卢塞恩（Ｌｕｃｅｒｎｅ）召开了重组人粒细胞巨噬细胞集烙刺激因子（ｒｈＧＭ－ＣＳＦ，商品名ｌｅｕｃｏｍａｘ，生白能）的国际研讨会，来自美国、瑞士...     

粒细胞-巨噬细胞集落刺激因子与神经保护

粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage clony-stimulating factor,GM-CSF)是一种血液细胞因子,与粒细胞和巨噬细胞生成有关.最近的研究提示,脑缺血后GM-CSF在神经系统的表达增高,提示其在脑缺血后神经保护中起着重要作用.GM-CSF主要是通过抗凋亡、促进侧支循环和抑制能量消耗来保护神经元.     

粒细胞-巨噬细胞集落刺激因子受体的研究进展

集落刺激因子（Colony Stimulating Factor）是一组控制粒细胞、单核一巨噬细胞和某些有关的造血细胞增殖和分化的糖蛋白，根据不同的分化阶段和造血祖细胞，可分为粒细胞-巨噬细胞集落刺激因子（GM—CSF）、粒细胞集落刺激因子（GCSF）、巨噬细胞集落刺激因子（M-CSF）以及多潜能集落刺激因子（Multi—CSF）叉称白细胞介素-3（IL-3）、红细胞生成素（Epo）、血小板生成素（Tpo）及干细胞因子（SCF）等。GM—CSF是一个多效性细胞因子，它促进多种造血细胞的存活，增殖、分化，     

粒细胞-巨噬细胞集落刺激因子治疗缺血性脑血管病

粒细胞-巨噬细胞集落刺激因子(granulocytc macrophage-colony stimulating factor,GM-CSF)是一种多功能生长因子,可刺激造血祖细胞(hemopoietic progenitor cell,HPC)增殖、分化和成熟并从骨髓向外周转移,诱导多种细胞增殖和分化.近年来的研究表明,GM-CSF在抗凋亡、诱导神经元分化和血管生成方面发挥着重要的作用,将是缺血性脑血管病治疗的新补充.文章就GM-CSF在缺血性脑血管病治疗中的作用做了综述.     

粒细胞-巨噬细胞集落刺激因子与不稳定型心绞痛的关系

目的　观察不稳定型心绞痛 (UAP)患者血清粒细胞 -巨噬细胞集落刺激因子 (GM- CSF)水平的变化 ,并进一步探讨其与血小板激活及内皮损伤之间的关系。方法　采用液相竞争放射免疫法检测 2 0例不稳定型心绞痛患者(U AP组 )、2 0例稳定型心绞痛患者 (SAP组 )及 2 0例正常对照组 (NC组 )血清 GM- CSF浓度 ,用酶联免疫双抗体夹心法测定其血浆血管性血友病因子 (v WF)及 P-选择素含量。结果　 U AP组血清 GM- CSF、血浆 P-选择素、v WF的水平明显高于 SAP组及正常对照组 (P均 <0 .0 1)。SAP组与对照组比较上述指标虽均升高 ,但无统计学意义 (P>0 .0 5 ) ;U AP患者血清 GM- CSF分别与血浆 P-选择素、v WF呈正相关 (r=0 .5 83及 r=0 .5 74 ;P均 <0 .0 1)。结论　不稳定型心绞痛患者血清 GM- CSF浓度升高 ,其可能与不稳定型心绞痛患者血小板激活和内皮损伤有关     

粒细胞-巨噬细胞集落刺激因子与支气管哮喘的研究进展

粒细胞一巨噬细胞集落刺激因子(GM-CSF)可以由支气管哮喘(简称哮喘)炎症部位的巨噬细胞、嗜酸粒细胞和上皮细胞产生.反之,GM-CSF也同样作用于巨噬细胞、嗜酸粒细胞、免疫细胞、中性粒细胞等.GM-CSF的高表达增强了机体对抗原的敏感性,导致气道炎症反应增加,在哮喘发病机制中起着重要作用.     

粒细胞-巨噬细胞集落刺激因子膀胱灌注治疗骨髓移植术后出血性膀胱炎的疗效观察

目的：评估粒细胞-巨噬细胞集落刺激因子(GM-CSF)治疗单倍体骨髓移植术后并发Ⅲ～Ⅳ级出血性膀胱炎的临床效果。方法：对7例出血性膀胱炎患者应用GM-CSF进行膀胱灌注300μg／d，连用5天。结果：7例患者中有6例完全缓解，1例无效，获得完全缓解的中位时间为11．3d(范围5～16d)。结论：膀胱灌注GM-CSF治疗骨髓移植术后出血性膀胱炎有效，未见不良反应发生。     

巨噬细胞集落刺激因子对DEL细胞增殖作用的研究

目的 研究巨噬细胞集落刺激因子对动脉粥样硬化病变处伴随血管平滑肌细胞共同积聚的巨噬细胞的增殖作用及机制。方法 以能分化为巨噬细胞表型的ＤＥＬ细胞为离体细胞实验模型。将富含巨噬细胞集落刺激因子的１９２９细胞条件培养液或重组的人巨噬细胞集落刺激因子加入体外培养的处于静止期的ＤＥＬ细胞,以氚标胸腺嘧啶核苷或５－溴－悄嘧啶 核苷掺入法分别检测ＤＥＬ细胞ＤＮＡ的合成,另以Ｎｏｒｔｈｅｒｎ ｂｌｏｔ法检测ＤＥ     

粒-巨噬细胞集落刺激因子在肺泡蛋白沉积症患者中的表达

目的 研究粒－巨噬细胞集落刺激因子（GM－CSF）蛋白及mRNA在4例肺泡蛋白沉积症（PAP）患者中的表达,探索PAP的发病机制。方法 采用ELISA法测定GM－CSF蛋白,采用RT－PCR检测外周血单个核细胞及肺泡巨噬细胞中GM－CSFmRNA表达水平。GM－CSFcDNA测序采用双脱氧链终止反应法。结果 4例PAP中3例外周血单个核细胞和肺泡巨噬细胞均检测不到GM－CSF释放,但mRNA表达水平均正常。cDNA测序发现1例PAP患者GM－CSFcDNA第382位碱基发生点突变（T→C）,致117位氨基酸序列发生改变（异亮氨酸→苏氨酸）。结论 GM－CSF蛋白表达异常与PAP发病可能有关,GM－CSFcDNA点突变可能是导致GM－CSF蛋白表达异常的原因之一。     

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。     

Serum neutralizing capacity of GM-CSF reflects disease severity in a patient with pulmonary alveolar proteinosis successfully treated with inhaled GM-CSF

Existence of anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) neutralizing antibody and treatment with recombinant GM-CSF are new topics in idiopathic pulmonary alveolar proteinosis (PAP). We have hypothesized inhaled GM-CSF is effective and neutralizing capacity of GM-CSF, not concentration of anti-GM-CSF antibody in serum reflect disease severity. A 57-year-old female smoker with idiopathic PAP was treated with inhaled GM-CSF. The response to the treatment was evaluated by diffusing capacity for carbon monoxide (DLCO), alveolar-arterial oxygen gradient ([A-a]DO2). Conventional serum markers, including KL-6, surfactant apoprotein (SP)-A, SP-D, carcino-embryonic antigen and cytokeratin fragment 19 (CYFRA), and concentration of anti-GM-CSF antibody were examined. The neutralizing capacity of GM-CSF in serum was evaluated using a GM-CSF dependent cell line, TF-1. Ground glass opacity disappeared at the end of the treatment. Her DLCO, [A-a]DO2 remarkably improved after treatment. The neutralizing capacity of GM-CSF declined in line with disease remission and it correlated significantly with DLCO (P = 0.0137). The concentration of anti-GM-CSF antibody had no significant relation with disease severity and serum markers including neutralizing capacity. Conventional serum markers other than CYFRA showed no significant correlation with Inhaled GM-CSF was effective for idiopathic PAR Serial measurement of neutralizing capacity of GM-CSF was useful to evaluate disease severity and the anti-GM-CSF antibody was proved to be a causative factor for PAR In the future, inhaled GM-CSF may replace whole lung lavage and response to GM-CSF and its optimal amount may be decided by the capacity.     

The potent antitumor effects of combined p16 gene and GM-CSF gene therapy through efficient induction of antitumor immunity

Purpose: Tumor suppressor gene therapy and cytokine gene therapy have limited antitumor effects when used alone. Thus, in the present study, we investigated the antitumor potentials of the combined transfer of the p16 tumor suppressor gene and the murine granulocyte-macrophage colony-stimulating factor (GM-CSF) gene. Methods: The adenovirus-harboring p16 gene (Adp16) and adenovirus-harboring GM-CSF (AdGMCSF) gene were utilized for the treatment of established tumors in vivo. The mice were inoculated s.c. with Renca renal carcinoma cells and 3 days later received an intratumoral injection of Adp16 in combination with AdGMCSF. Results: The results demonstrated that tumor-bearing mice treated with Adp16 and AdGMCSF showed more potent inhibition of tumor growth and a prolonged survival period than mice treated with Adp16, AdGMCSF, adenovirus-expressing β-galactosidase or PBS (P < 0.01). Treatments of the mice with Adp16 alone or AdGMCSF alone also showed obvious antitumor effects as compared with those mice treated with PBS (P < 0.05). After combined p16 and AdGMCSF gene therapy, the expression of H-2K^d and Fas molecules on freshly isolated tumor cells increased markedly, and more CD₄ ⁺ T cells and CD₈ ⁺ T cells infiltrated in the tumor sites. The cytotoxicity of natural killer cells and specific cytotoxic T lymphocytes increased more significantly after the combined therapy. Conclusions: Our results demonstrated that combination p16 gene and GM-CSF gene therapy could inhibit the growth of established tumors in mice more significantly through efficient induction of antitumor immunity. Received: 11 April 2000 / Accepted: 19 July 2000     

Bone marrow myeloid cell kinetics during treatment of small cell carcinoma of the lung with chemotherapy not associated and associated with granulocyte-macrophage colony-stimulating factor

Summary Information on the kinetics of bone marrow (BM) myeloid precursors (BMMP) is required for integrating cancer chemotherapy with granulocyte-macrophage colony-stimulating factor (rhGM-CSF), with the aim of reducing neutropenia. Using bivariate flow-cytometric analysis of the in vivo incorporation of bromodeoxyuridine (BUDR) vs DNA content we have studied the kinetics of BMMP in 21 patients with SCLC during the first of six chemotherapy courses (etoposide, epirubicin, andcis-platinum, days 1–3, every 21 days), given alone (eight patients) or followed by rhGM-CSF (10g/kg/day s.c, days 4–14) as BM rescue (eight patients) or both preceded (days -17 to -7, as BM priming) and followed by rhGM-CSF (five patients). At 11–14 days after the start of these therapies there was an increase in the baseline proliferative activity of proliferating BMMP and a shortening in the time needed by the metamyelocyte to mature and to leave the marrow. Both effects were greater and were maintained to a significantly greater degree a week later in patients who received chemotherapy plus rhGM-CSF rescue than in those who received chemotherapy alone or rhGM-CSF priming alone. At day 11–14 the pretreatment median cell production rate of pBMMP was increased by 340%, 150%, and 183% and the maturation time was reduced by 80%, 45%, and 57%, respectively, in the three groups. A week later, the corresponding figures were 206%, 111%, and 157% and 50%, 18%, and 45%. Hence, an identical rhGM-CSF schedule is more effective in increasing the neutrophil production by BMMP when given following chemotherapy as BM rescue than before it as BM priming. In both the rescue and the priming schedule, the increase in proliferative activity of BMMP just at the end of rhGM-CSF stimulation was linked to both an increase in the labeling index and a reduction in duration of S-phase (TS), while a week later it was linked solely to reduction in TS. This could actually reduce one of the two kinetic targets of subsequently administered cytostatics, i.e., a high LI and a long time spent in S phase. From this study, accurate kinetic data can be obtained with the in vivo BUDR technique that are useful in scheduling rhGM-CSF.     

人重组粒细胞集落刺激因子动员的外周血采集物和稳态骨髓采集物免疫学特性的对比研究

目的研究人重组粒细胞集落刺激因子(rhG-CSF)动员的健康供者外周血采集物(G-PB)和稳态骨髓采集物(SS-BM)在免疫学特性方面的异同。方法对2003年4月至10月北京大学血液病研究所接受异基因造血干细胞移植的15名亲缘供者,采用流式细胞仪测定两种移植物的T细胞亚群、树突细胞(DC)亚群、单核细胞及共刺激分子CD28的表达,并用四甲基偶氮唑盐(MTT)法和酶联免疫吸附法(ELISA)检测G-PB和SS-BM中T淋巴细胞增殖能力和白细胞介素-4(IL-4)、干扰素-γ(IFN-γ)的分泌情况。结果G-PB中淋巴细胞,CD3+、CD3+CD4+、CD3+CD8+T细胞,单核细胞的含量以及CD4与CD8比值均高于SS-BM(P<0.01);G-PB的T细胞增殖能力与SS-BM相比差异无显著性意义(P>0.05);每微升G-PB移植物中T细胞分泌细胞因子IFN-γ、IL-4的量均高于SS-BM(P<0.05、P<0.01),G-PB中IL-4与IFN-γ比值与SS-BM比较差异无显著性意义;G-PB中DC1、DC2的含量高于SS-BM(P<0.01),DC2与T淋巴细胞的比值低于SS-BM(P<0.01),单核细胞和T淋巴细胞的比值显著高于SS-BM(P<0.01);CD3+、CD4+、CD8+细胞上CD28表达的百分比和总体表达均高于SS-BM(P<0.01);CD8+CD28-T细胞与T淋巴细胞的比值和SS-BM的差异无显著性意义(P>0.05)。结论G-PB与SS-BM在免疫特性方面的异同可能是二者在移植后具有相似的急性移植物抗宿主病(GVHD)发生率以及G-PB移植后较高的慢性GVHD发生率和较低复发率的免疫学基础。     

AA和MDS患者骨髓CD+34细胞表面GM-CSFR的表达及意义

目的 探讨再生障碍性贫血（AA）和骨髓增生异常综合征（MDS）患者骨髓CD34^＋细胞表面粒细胞-巨噬细胞集落刺激因子受体（GM—CSFR）的表达情况。方法 用流式细胞术（FCM）和单克隆抗体技术检测14例AA、23例MDS和8例非血液病患者骨髓CD34^＋细胞表面的GM—CSFR表达率。结果 MDS组骨髓CD34^＋细胞表面GM-CSFR的表达率明显高于对照组及AA组（P〈O．05）,而AA组与对照组间差异无统计学意义（P〉0．05）。结论 AA患者造血干细胞没有质的缺陷;造血干细胞异常可能是MDS的主要发病机制之一。     

丝裂霉素诱导凋亡癌细胞致敏树突状细胞后的免疫应答

目的 观察树突状细胞(DC)从丝裂霉素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 用粒细胞-巨噬细胞集落刺激因子(GM-CSF)加IL-4从人外周血分化、诱导DC,丝裂霉素在体外诱导培养的人胆管癌细胞凋亡,将DC、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DC、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的DC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淋巴细胞特异性地杀伤胆管癌细胞。 结论 丝裂霉素诱导凋亡癌细胞可以致敏rhGM-CSF加rhIL-4从人外周血单个核细胞诱导、扩增出的DC,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。     

Ratio of complement receptor over Fc-receptor III expression: A sensitive parameter to monitor granulocyte-macrophage colony-stimulating factor effects on neutrophils

Summary In vitro activation of human granulocytes leads to altered expression of distinct surface antigens. Compared with the changes observed with classic activating reagents such as the phorbol ester PMA similar, but less pronounced alterations of surface antigen expression were observed upon granulocyte activation with human recombinant granulocyte-macrophage colony-stimulating factor (hrGM-CSF). In particular, stimulation with hrGM-CSF is followed by an enhanced expression of the complement receptors CD35 (CR1) and CD11b (CR3) while the low affinity Fc- receptor CD16 (FcRIII) is downregulated. In order to investigate whether there are similar effects under in vivo conditions, we studied the granulocytes from patients undergoing rhGM-CSF therapy before, during, and after treatment. We found a marked increase in CD35 (CR1) and CD11b (CR3) expression and a substantial decrease or even loss of CD16 (FcRIII) on these granulocytes. These changes correlated well with our in vitro data and occurred extremely rapidly after therapy onset. Furthermore, therapy monitoring using ratios calculated by the mean fluorescence channel numbers of CR and FcRIII stainings may combine the advantage of high sensitivity with high reproducibility as a result of the contrasting change in CR and FcRIII expression during granulocyte activation. Being nonparametric values, such ratios are not influenced by individual flow cytometry standardization. Taken together, these activation-associated changes of surface receptor expression and especially of CR over FcRIII ratios are useful parameters for monitoring the in vivo effects of rhGM-CSF.