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。      

自体恶性黑色素瘤抗原激活的树突状细胞体外诱导免疫试验

树突状细胞(dendritic cells,DC)是迄今为止所知的抗原呈递能力最强的抗原呈递细胞(antigen presenting cells,APC).肿瘤患者因DC免疫功能低下致宿主抗肿瘤免疫无能.研究表明,人外周血单核细胞在体外用粒/巨噬细胞集落刺激因子(granulocyte/macrophage colony-stimulating factor,GM-CSF)及白介素4(IL-4)诱导后,可培养出表达高水平MHCI、MHCII类抗原和CD80、CD86等因子的DC.本实验拟应用黑色素瘤患者外周血单核细胞,经GM-CSF及IL-4诱导后获得DC,再经自体肿瘤细胞冻融物冲击,在体外激活自体T淋巴细胞产生细胞毒性T淋巴细胞(CTL),观察其体外抗肿瘤作用.     

粒细胞集落刺激因子及其受体在肺癌中表达与作用的研究进展

粒细胞集落刺激因子(granulocyte colony-stimulating factor,G-CSF)被广泛应用于治疗肿瘤放、化疗所致的白细胞减少.研究发现,多种实体肿瘤中有G-CSF及粒细胞集落刺激因子受体(G-CSF receptor, G-CSFR)的表达,且G-CSF可通过G-CSF/G-CSFR的自分泌/旁分泌通路与环氧化酶通路等影响肿瘤细胞增殖、凋亡和微血管形成,但其确切机制尚需要进一步研究.本文就G-CSF 及G-CSFR在肺癌中的表达及其作用机制的研究进展作综述.     

JAK-STAT信号通路与巨噬细胞炎症调控的研究进展

JAK-STAT信号通路是近年来备受关注的一条细胞因子信号转导通路，在细胞的增殖、凋亡、分化和免疫应答中发挥重要调控作用。巨噬细胞是机体内最主要的炎症效应细胞，在不同的环境和因素刺激下，可分化为促炎和抗炎两种类型，并参与多种疾病的发生发展过程。然而，在炎症因子刺激下JAK-STAT信号是如何激活的，JAK-STAT又是如何调控巨噬细胞极化分型和炎症走向，其机制并不明确。本文综述了JAK-STAT与巨噬细胞炎症反应调控的最新进展，旨在阐明JAK-STAT在巨噬细胞炎症调控中的关键作用，以期为机体炎症疾病的防治提供新的依据和策略。     

JAK-STAT信号通路与巨噬细胞炎症调控的研究进展

JAK-STAT信号通路是近年来备受关注的一条细胞因子信号转导通路，在细胞的增殖、凋亡、分化和免疫应答中发挥重要调控作用。巨噬细胞是机体内最主要的炎症效应细胞，在不同的环境和因素刺激下，可分化为促炎和抗炎两种类型，并参与多种疾病的发生发展过程。然而，在炎症因子刺激下JAK-STAT信号是如何激活的，JAK-STAT又是如何调控巨噬细胞极化分型和炎症走向，其机制并不明确。本文综述了JAK-STAT与巨噬细胞炎症反应调控的最新进展，旨在阐明JAK-STAT在巨噬细胞炎症调控中的关键作用，以期为机体炎症疾病的防治提供新的依据和策略。     

重组人粒细胞巨噬细胞集落刺激因子治疗羟基脲致皮肤顽固性溃疡1例报道

目的:观察重组人粒细胞巨噬细胞集落刺激因子(recombinant human granulocyte/macrophage colony-stimulating factor(rhGM-CSF))治疗慢性粒细胞白血病服羟基脲导致皮肤顽固性溃疡的疗效,探讨rhGM-CSF促进溃疡愈合的作用机制。方法:对常规治疗与在常规治疗基础上局部应用rhGM-CSF进行比较,观察溃疡愈合情况。结果:局部应用rhGM-CSF疗效明显。结论:rhGM-CSF能促进溃疡愈合。     

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。     

造血细胞因子临床应用

本文对几种造血细胞因子，包括重组人粒／巨噬细胞集落刺激因子(Recombinant human granulocytemacropbage colony-stimulating factor，rhGM-CSF)、重组人粒细胞集落刺激因子(Recombinant human granulocyte colony-stimulating factor。rhG-CSF）、重组人红细胞生成素（Recombinant human erythropoietin，rhEPP)-干扰素(Interferon，IFN)、白介素-2(interleukin-2）与LAK细胞(Lymphokine activated killer cell)的临床应用机理、适应症、毒副作用，根据我们几年来的实践体会．结合国内外文献复习作了介绍。近年细胞因子的研究进展迅速，新的细胞因子不断被发现并推向临床，对已用于临床的细胞因子的适应症、用药时机、剂量和毒副作用的防治等临床经验也越来越丰富。用细胞因子治疗一些过去被认为难治的疾病已展示出可喜的前景，但目前也还存在一些问题，如价格昂贵、市场品种繁多、效应不一等等，给临床应用带来困难。     

皮下注射粒细胞-巨噬细胞集落刺激因子预防多发性骨髓瘤患者侵袭性真菌病效果分析

目的探讨皮下注射粒细胞-巨噬细胞集落刺激因子(GM-CSF)对多发性骨髓瘤(MM)患者侵袭性真菌病(IFD)的预防效果。方法回顾性分析2015年1月至2021年6月哈尔滨医科大学附属第二医院收治的接受诱导化疗的222例初治MM患者的临床资料。患者均于中性粒细胞(ANC)≤1.5×109/L时应用GM-CSF(3～5 μg·kg-1·d-1, GM-CSF组)或粒细胞集落刺激因子(G-CSF, 2～5 μg·kg-1·d-1, G-CSF组), 白细胞计数(WBC)≥10.0×109/L时停用。比较两组患者IFD(包括确诊、临床诊断和拟诊)及突破性侵袭性真菌感染发生情况。结果所有患者中IFD发生率为8.1%(18/222)。GM-CSF组和G-CSF组IFD发生率分别为3.5%(3/85)和10.9%(15/137), 两组差异有统计学意义(χ2=3.88, P=0.049)。GM-CSF组接受真菌感染预防的9例及G-CSF组接受真菌感染预防的15例患者中, 发生突破性侵袭性真菌感染分别为0、7例, 两组差异有统计学意义(P=0.022)。结论 MM患者应用GM-CSF可减少IFD和突破...     

肿瘤化疗联合粒细胞-巨噬细胞刺激因子的辅助治疗

正多年来,粒细胞刺激因子(G-CSF)和粒细胞-巨噬细胞刺激因子(GM-CSF)广泛应用于治疗放、化疗后的骨髓抑制,及骨髓移植、再生障碍性贫血、骨髓增生异常综合征等相关的白细胞减少症。本文主要综述肿瘤化疗后联合使用GM-CSF辅助治疗骨髓抑制、真菌感染、口腔黏膜炎及免疫抗肿瘤     

Priming effects of GM-CSF, IFN-gamma and TNF-alpha on human neutrophil inflammatory cytokine production

Identifying and evaluating the priming agents for cytokine release by neutrophils might be helpful in controlling the innate immune response of the host. In the present study we examined the role of granulocyte/macrophage colony-stimulating factor (GM-CSF), interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) as priming agents for interleukin (IL)-1beta, IL-6 and TNF-alpha production by stimulated neutrophils from control subjects and malignant melanoma patients. When the cells from controls and patients were preincubated with primer agents, opsonized zymosan-stimulated inflammatory cytokine production was enhanced. The major neutrophil-priming factor for IL-6 secretion by polymorphonuclear leukocytes (PMNs) in the control and patient groups was TNF-alpha. However, GM-CSF and IFN-gamma are also significant primers. GM-CSF priming was critical for the release of TNF-alpha from PMNs in control and melanoma patients. The ability of GM-CSF, IFN-gamma and TNF-alpha to serve as effective priming agents for inflammatory mediator production by PMNs revealed a new role for these cytokines in the innate immune response of the melanoma-bearing host.     

Clinical phase I intratumoral administration of two recombinant ALVAC canarypox viruses expressing human granulocyte-macrophage colony-stimulating factor or interleukin-2: the transgene determines the composition of the inflammatory infiltrate

Immunotherapy employs cytokines for modifying local inflammatory reactions. Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been shown to activate dendritic cells, macrophages, and granulocytes leading to clinical trials using GM-CSF-based cancer vaccine approaches. Interleukin-2 (IL-2) is an important T cell stimulatory cytokine approved as exogenous antitumor agent. The ALVAC viral vector system uses a recombinant canarypox virus for local gene expression. We report a phase I clinical trial using intratumoral administration of ALVAC GM-CSF or ALVAC IL-2 in skin metastases of melanoma or leiomyosarcoma. ALVAC GM-CSF and ALVAC IL-2 were injected at 107.12 and 106.92, 50% cell culture infectious dose in eight metastases with acceptable tolerability. Local and systemic inflammatory reactions were observed. The transgene determined the local infiltrate: GM-CSF induced monocyte and macrophage enrichment of the peritumoral inflammatory infiltrate, whereas IL-2 increased local T lymphocytes. Stable disease of injected lesions was seen after ALVAC GM-CSF application, whereas ALVAC IL-2 treatment led to partial regression in three out of eight injected tumors, accompanied by decreased expression of melanocytic antigens. Local GM-CSF expression could be induced. In summary, ALVAC GM-CSF and ALVAC IL-2 injections are safe and can mediate local biologic and immunologic effects.     

Recombinant human TNF-alpha stimulates the secretion of granulocyte colony-stimulating factor in vivo.

Tumor necrosis factor (TNF) is a macrophage-derived cytokine that causes hemorrhagic necrosis of several human tumors in vitro. It has a wide range of biologic effects including stimulation of secretion of both granulocyte colony-stimulating factor (G-CSF) and granulocyte/macrophage colony-stimulating factor (GM-CSF) by normal adult lung fibroblasts in culture. No in vivo data are available on the effect of exogenously administered TNF on cytokine production. In the studies reported here, we show that G-CSF accumulates in the serum in vivo in response to recombinant TNF (rTNF) administration. At the peak of the response circulating levels of 2-6 ng/ml of biologically active G-CSF are detectable. Surprisingly, circulating levels of GM-CSF, interleukin-3 as well as a number of other cytokines were not detectable within the limits of the assays. The results indicate that the levels of GM-CSF or interleukin-3 are minimally 100-fold lower than the peak levels of G-CSF. These data illustrate the complex interplay that cytokines have in vivo. Understanding these interactions in humans is crucial to the correct use of this new class of agents in the clinic.     

Identification of colony-stimulating factor activity in patients with malignant tumors associated with excessive leukocytosis.

We have tried to demonstrate and identify colony-stimulating factor (CSF) activity in the plasma, pleural fluid, ascites or culture supernatant of tumor cells in 11 patients with malignant tumors associated with unexplained persistent leukocytosis. The specimens were treated with anti-granulocyte (G)-CSF or anti-granulocyte/macrophage (GM)-CSF monoclonal antibodies, then added to GM-progenitor (CFU-GM) cultures without exogenous CSFs. In all patients, untreated specimens generated CFU-GM-derived colonies, and colony formation was clearly inhibited by only one of the two antibodies, indicating the presence of either G-CSF or GM-CSF in the specimens. Furthermore, we measured the concentrations of G-CSF or GM-CSF in the specimens using an enzyme-linked immunosorbent assay, and confirmed the results by CFU-GM assay. Two patients were shown to have GM-CSF-producing tumors, while the other patients were G-CSF-producing. These assays are useful in identifying CSF activity in patients with CSF-producing tumors.     

Granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor promote malignant growth of cells from head and neck squamous cell carcinomas in vivo

Granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) are used to ameliorate cancer therapy-induced neutropenia and mucositis. Yet, first data in head and neck squamous cell carcinoma (HNSCC) indicate an impaired long-term prognosis on G-CSF treatment, and previous studies showed a contribution of both factors to the progression of human epithelial tumors. Therefore, we investigate the role of G-CSF and GM-CSF in progression of tumor cells from human HNSCC. Both factors stimulated proliferation and migration of tumor cell lines established from patient tumors expressing G-CSF and GM-CSF and/or their receptors. Blockade of G-CSF and GM-CSF inhibited tumor cell invasion in a three-dimensional organotypic culture model. The contribution of both factors to tumor malignancy was further confirmed in nude mouse transplants in vivo. Invasive and malignant growth yielding a similar tumor phenotype as the original patient tumor was exclusively observed in G-CSF- and GM-CSF-expressing tumors and was associated with enhanced and persistent angiogenesis and enhanced inflammatory cell recruitment. Although factor-negative tumors grew somewhat faster, they were characterized by lack of invasion, reduced and transient angiogenesis, and large necrotic areas. These data provide evidence for a progression-promoting effect of G-CSF and GM-CSF in human HNSCC and suggest further detailed evaluation of their use in the therapy of these tumors.     

Hematopoietic growth factors: biology and clinical application.

A number of recently identified cytokines have been implicated in the development of blood cells and their functional activation. These include granulocyte colony-stimulating factor (G-CSF), granulocyte/macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), and interleukins-1, -3, and -6. The two that have been studied most extensively clinically are G- and GM-CSF. Biologic differences between these two agents have important implications for their use in particular clinical settings. Whereas G-CSF for the most part has demonstrated lineage specificity in stimulating production of neutrophil granulocytes, GM-CSF stimulates production of all types of granulocytes and is a potent activator of monocytes and macrophages as well. In addition, GM-CSF has been found to induce other cytokines, such as tumor necrosis factor and interleukin-1. Functional differences such as influencing neutrophil migration have also been noted; GM-CSF can be a potent inhibitor of neutrophil migration. The interaction of these and other cytokines in inflammation and injury is the determinant of the ultimate clinical outcome. Potential clinical applications for these growth factors are discussed, and specific clinical studies during which recombinant human G-CSF was used are reviewed.     

Sustained expression of the pim-1 kinase is specifically induced in myeloid cells by cytokines whose receptors are structurally related.

We have examined the effects of myeloid growth factors on expression of the pim-1 kinase protein in human and murine myeloid cells. pim-1 protein was identified in K562 cells by immunoblotting as a 33 kDa protein. In the human factor-dependent myeloid leukemia cell line M07E, pim-1 protein was induced by interleukin 3 (IL-3) or granulocyte-macrophage colony-stimulating factor (GM-CSF), with maximum expression by 4 h. Expression continued for the duration of growth factor exposure, but declined rapidly when cytokines were removed. GM-CSF induced pim-1 protein in a dose-dependent manner, with expression being proportional to the proliferative effect of the cytokine. To examine the specificity of pim-1 protein induction, we compared pim-1 protein levels in myeloid cells which demonstrated different GM-CSF response phenotypes. We also examined the effects on pim-1 protein expression of different growth factors which induced similar response phenotypes. GM-CSF induced pim-1 protein in several myeloid cell lines, most of which demonstrated a proliferative response, but did not induce pim-1 protein expression in neutrophils or monocytic cells. In contrast, the murine cell line Mac-11 expressed pim-1 message in response to IL-3 and GM-CSF, but not in response to bryostatin or M-CSF, which were equivalent mitogens. In human U937 myeloid cells sustained expression of pim-1 protein was induced by GM-CSF, G-CSF and IL-6, but not by bryostatin. Expression of the pim-1 kinase protein in response to myeloid cytokines depends on both the nature of the growth factor and the response phenotype. The pim-1 kinase may be an important intermediate in transmembrane signaling or response phenotype induced by IL-3, GM-CSF and other cytokines whose receptors are structurally similar. Its constitutive expression in some myeloid leukemia cell lines suggests activation of signal cascades utilized by myeloid growth factors.     

Effect of human recombinant granulocyte/macrophage colony-stimulating factor and native granulocyte colony-stimulating factor on clonogenic leukemic blast cells

The effects of human recombinant granulocyte/macrophage colony-stimulating factor (GM-CSF) and human native purified granulocyte colony-stimulating factor (G-CSF) on the growth of clonogenic leukemic blast cells from eight Japanese patients with acute myeloblastic leukemia were studied, using an in vitro leukemic blast colony assay. The results showed that GM-CSF stimulated leukemic blast colony formation in all cases examined, whereas G-CSF stimulated colony formation in four of the eight cases. The maximum stimulating activity of GM-CSF on the growth of clonogenic leukemic blast cells was higher than that of G-CSF in the majority of cases, while sometimes GM-CSF and G-CSF worked synergistically. Thus, the clonogenic leukemic blast cell populations seemed to be heterogeneous with respect to their in vitro response to growth regulators.     

Blood and bone marrow eosinophilia in malignant tumors. Role and nature of blood and tissue eosinophil colony-stimulating factor(s) in two patients

Eosinophilia of the blood and bone marrow may be encountered in patients with disseminated malignancies. It is unrelated to the histologic type of the tumor but usually reflects its aggressiveness and prognostic outlook. Its pathogenesis is controversial. The presence of eosinophil colony-stimulating factor(s) in serum and malignant tissue extracts was evaluated in two cases of malignancies accompanied by marked blood and bone marrow eosinophilia. When compared with controls, serum and tumor tissue extracts stimulated the growth of G, M, and Eo colonies in semisolid cultures of human bone marrow. Such stimulating effect was inhibited by the addition of antibodies to granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3. Thus, the colony-stimulating factor(s) play a role in the pathogenesis of the eosinophilia associated with some malignant tumors, and these factors appear to include GM-CSF, interleukin-3, and probably others.     

Growth regulation of the AML-193 leukemic cell line: evidence for autocrine production of granulocyte-macrophage colony-stimulating factor (GM-CSF), and inhibition of GM-CSF-dependent cell proliferation by interleukin-1 (IL-1) and tumor necrosis factor (TNF alpha)

The human leukemic cell line AML-193 was tested for its proliferative response to endogenously produced autocrine factors and to a variety of cytokines and colony-stimulating factors. Cells grown in the absence of GM-CSF incorporated tritiated thymidine, and this was partially reversed by adding neutralizing anti-GM-CSF antibodies to the culture medium, suggesting that it was due, at least in part, to autocrine GM-CSF production. This was confirmed by immunopurification of a GM-CSF-like activity from cell supernatant of AML-193 cells grown in serum free medium in the absence of exogenous GM-CSF. When AML-193 cells were cultured with GM-CSF in combination with other cytokines, Interleukin-1 alpha and beta (IL-1 alpha and beta), Interleukin-3 (IL-3), Interleukin-6 (IL-6), granulocyte colony-stimulating factor (G-CSF) and tumor necrosis factor alpha (TNF alpha), none of them affected the concentration of GM-CSF required to induce 50% of maximum proliferation (D50). However, the maximum proliferation induced by GM-CSF alone was drastically decreased by IL-1 alpha, IL-1 beta and TNF alpha. Inhibition caused by exposure of the AML-193 to IL-1 for up to 24 hr was reversible, ruling out a direct cytotoxic effect.