Agranulocytosis induced by antithyroid therapy: effects of treatment with granulocyte colony stimulating factor

A 26-year-old woman was admitted to hospital with high fever, severe tonsillitis, and gastroenteritis. Because of Graves' disease she had been treated with methimazole for 18 months. Leukopenia and agranulocytosis in combination with a typical bone marrow, exhibiting a complete arrest of myelopoiesis at the stage of promyelocytes led to the diagnosis of an antithyroid therapy induced agranulocytosis. After 1 week of antibiotic treatment without changes in neutrophil counts, granulocyte colony stimulating factor treatment at a dose of 300 g/day subcutaneously was started. Twenty-four hours after the first administration the neutrophil counts began to rise, to 4389/l, with a maximum after the third administration and stabilizing at normal levels within 10 days. Since agranulocytosis is considered to be a severe and fatal complication of methimazole therapy, treatment with granulocyte colony stimulating factor seems to be useful for this life-threatening condition.Abbreviations ANC absolute neutrophil count - G-CSF granulocyte colony stimulating factor - GM-CSF granulocyte macrophage colony stimulating factor     

Ultrastructural effects of granulocyte colony stimulating factor (G-CSF) and macrophage colony stimulating factor (M-CSF) on rat neutrophils and monocytes

Human granulocyte colony stimulating factor (G-CSF) and macrophage colony stimulating factor (M-CSF) were administered intravenously to rats, and their effects on neutrophils and monocytes were examined by electron microscopy. G-CSF increased the number of cytoplasmic granules in neutrophils. It also enhanced maturation of the nuclear shape in the neutrophils, while chromatin condensation and peroxidase distribution remained immature. M-CSF induced proliferation of monocytes in peripheral blood and bone marrow, but did not affect morphology or distribution of peroxidase reactivity. This study was presented at the 25th Annual Meeting of the Clinical Electron Microscopy Society of Japan, Matsumoto, September 28–30, 1993.     

Urinary bladder carcinoma producing granulocyte colony stimulating factor (G-CSF): A case report with immunohistochemistry

A rare case of urinary bladder carcinoma with granulocyte colony stimulating factor (G-CSF) production was reported. In an 83-year-old female, marked neutrophilia in the peripheral blood decreased from 132,500/mm³ to 3,300/mm³ after tumour resection. The tumour was a transitional cell carcinoma. The serum G-CSF level reduced from 238 pg/ml pre-operatively to normal (60 pg/ml) after the operation. Immunohistochemical investigation of the resected tumour with monoclonal antibody specific for G-CSF revealed positive staining in the carcinoma cells, confirming G-CSF secretion.     

白介素-1和粒细胞集落刺激因子对胎膜早破合并羊膜腔感染预测的意义

目的评价白介素-1(IL-1)和粒细胞集落刺激因子(G-CSF)在预测无症状胎膜早破合并羊膜腔感染中的作用。方法检测72例无症状胎膜早破患者(研究组)血清白介素-1和粒细胞集落刺激因子含量,并与同期72例正常妊娠(对照组)比较。结果研究组血清白介素-1和粒细胞集落刺激因子含量均高于对照组,差异有显著性(152.37±37.86 pg/ml vs 80.52±29.90 pg/ml,109.28±51.67 vs 80.51±23.39,P<0.05)。结论研究组血清白介素-1和粒细胞集落刺激因子含量均较对照组升高,白介素-1和粒细胞集落刺激因子可作为无症状胎膜早破合并羊膜腔感染的预测因子。     

粒细胞集落刺激因子调节T淋巴细胞免疫功能的实验研究

目的研究粒细胞集落刺激因子(G-CSF)对T淋巴细胞免疫功能的调节作用。方法以15例健康供者为对象,在其应用G-CSF前后,用流式细胞仪(FCM)检测外周血T淋巴细胞表面免疫标记、Annexin V及细胞内细胞因子的表达,用淋巴细胞转化试验检测T淋巴细胞的增殖反应。结果供者用G-CSF后,CIM^ 、CD8^ 细胞数无明显变化,CIM^ 、CD8^ 细胞内IFN-γ表达降低,IL-4表达增高;T细胞对丝裂原的增殖反应降低。Th1和Th2细胞表面Annexin V的表达无显著性差异。结论G-CSF制了丝裂原刺激T细胞的功能,使供者T细胞向Th2型细胞转化,细胞凋亡不是Th1／Th2转化的机制。     

Granulocyte colony stimulating factor attenuates hyperoxia-induced lung injury by down-modulating inflammatory responses in neonatal rats

Purpose

Granulocyte colony stimulating factor (G-CSF) has been known to increase neutrophil production and have anti-inflammatory properties, but the effect of G-CSF on pulmonary system is in controversy. We investigated whether G-CSF treatment could attenuate hyperoxia-induced lung injury, and whether this protective effect is mediated by the down-modulation of inflammatory responses in a neonatal rat model.

Materials and Methods

Newborn Sprague-Dawley rats (Orient Co., Seoul, Korea) were subjected to 14 days of hyperoxia (90% oxygen) beginning within 10 h after birth. G-CSF (20 µg/kg) was administered intraperitoneally on the fourth, fifth, and sixth postnatal days.

Results

This treatment significantly improved hyperoxia-induced reduction in body weight gain and lung pathology such as increased mean linear intercept, mean alveolar volume, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling positive cells. Hyperoxia-induced activation of nicotinamide adenine dinucleotide phosphate oxidase, which is responsible for superoxide anion production, as evidenced by upregulation and membrane translocation of p67^phox was significantly attenuated after G-CSF treatment, as were inflammatory responses such as increased myeloperoxidase activity and mRNA expression of transforming growth factor-β. However, the attenuation of other proinflammatory cytokines such as tumor necrosis factor-α and interleukin-6 was not significant.

Conclusion

In sum, G-CSF treatment significantly attenuated hyperoxia-induced lung injury by down-modulating the inflammatory responses in neonatal rats.     

集落刺激因子的生物功能

正常人体中,只有骨髓中存在多能干细胞,它能产生血液循环中所有的成熟红细胞,中性粒细胞,嗜酸性粒细胞,嗜碱性粒细胞,单核细胞,淋巴细胞及血小板。这些成熟细胞的寿命较短,它们持续不断的更替有赖于多能干细胞的大量增殖。血细胞的产生受到复杂而严密的调控,保证健康状态的血细胞水平维持稳定,并在受到外伤或感染时能迅速变化。每个成年人每日大约更新１２００亿个粒细胞,若感染时,其产生数量最少增加十倍。 １９６６年, Ｂｒａｄｌｙ, Ｍｅｔａｃａｆｆ和Ｉｃｈｉｋａｗａ等建立了一种细胞培养系统,允许粒细胞和单核细胞的…     

Lung squamous cell carcinoma producing both parathyroid hormone-related peptide and granulocyte colony stimulating factor

An autopsy case of a 61 year old male with primary squamous call carcinoma of the lung with associated marked leukocytosls and hypercalcemla Is reported. High levels of serum parathyroid hormone-related peptide (PTHrP) and granulocyte colony stimulating factor (GCSF) were detected. The tumor cells distinctly showed positive cytoplasmic knmunoreactions with anti-PTHrP and anti-GCSF antibodies. Marked granulocytosls and thin bony trabeculae lacking osteoblasts were observed in the vertebral bone. Calcium deposits were found In the proximal tubules of the kidneys. Infarcts were seen as a result of fibrin thrombosis of the splenic artery. The tumor was successfully transplanted into nude mice in which the high levels of serum PTHrP and GCSF were reproduced. These results indicate that the tumor simultaneously produced both PTHrP and GCSF causing the paraneoplastic syndromes of hypercalcemia and ieukocytosis.     

Divergent regulation of cell surface protease expression in HL-60 cells differentiated into macrophages with granulocyte macrophage colony stimulating factor or neutrophils with retinoic acid

Taking advantage of the recently demonstrated presence of N-aminopeptidasesand the serine protease dipeptidyi aminopeptidase IV (DPP IV)at the surface of human myeloblastic HL-60 cells, the regulationof these protease activities in HL-60 cell differentiation hasbeen assessed using combined spectrophotometric and flow cytometricassays. Addition of human recombinant granulocyte macrophagecolony stimulating factor (rHu-GM-CSF) to HL-60 cells to inducedifferentiation into macrophages led to a time and dose-dependentincrease in both cell surface N-aminopeptidase and DPP IV activities.Protease up-regulation was due to an enhancement in cell surfaceprotease number, associated with a slight rise in apparent affinitiesof the enzymes for their substrates. In contrast, in HL-60 cellsinduced to differentiate into neutrophils in the presenceofretinoic acid, expression of cell surface N-amlnopeptidaseswas almost completely abolished in a time-and dose-dependentfashion, and this down-regulation was accompanied by a weakbut significant decrease in affinity. However, no noticeabledifference was seen in serine DPP IV expression between retinoicacid-treated and untreated HL-60 cells. Retinoic acid treatmentalso reduced soluble protease activity in vitro indicating thatdown-regulation of membrane aminopeptldases was not due to theirproteolytic clip. No modulation in the activity of any of theenzymes tested was seen with human recombinant tumor necrosisfactor- or retinol which do not induce HL-60 cell differentiation.The up-regulation of cell surface protease expression in HL-60cells differentiated into macrophages was similar to that observedin monocytes isolated from peripheral blood: both DPP IV andN-aminopeptidase activities strictly increased on cells thatundergo macrophage maturation (up to 5-fold) and independentlyof the nature of the differentiation inducer. Thus, the distinctivepatterns of N-aminopeptidase and DPP IV expression that areseen in differentiating neutrophils and macrophages appear tobe relatedto differences in stage of myeloid maturation. Becausecell surface proteases are crucially involved in leukocyte functions,the data presented suggest that alterations in cell surfaceprotease expression are associated with events controlling thedifferentiation of immature cells.     

联合应用粒细胞集落刺激因子和骨髓基质干细胞治疗急性心肌缺血的实验研究

目的： 探讨粒细胞集落刺激因子和自体骨髓基质干细胞移植共同应用对心肌缺血的治疗作用。方法: 采用自体骨髓基质干细胞（MSCs）在体外培养扩增。在结扎冠状动脉造成急性心肌缺血后，把被5溴-2脱氧尿苷（BrdU）标记后的MSCs移植到自体的缺血心肌中，同时腹腔注射粒细胞集落刺激因子5 d。4周后通过激光共聚焦显微镜验证移植后的MSCs是否向心肌细胞分化，通过心脏彩超和多导生理记录仪测定缺血心肌在移植自体MSCs后是否能提高心功能。另外，用Masson 氏3色染色法从左室中断面切片量化心肌梗死范围。结果: 移植4周后，MSCs向肌细胞分化，表达出α-横纹肌肌动蛋白（α-sarcomeric actin）和存在于心肌闰盘中的connexin 43，粒细胞集落刺激因子和自体骨髓基质干细胞移植结合应用的治疗组，左心室收缩功能明显强于单独应用自体骨髓基质干细胞移植的治疗组，移植后心肌左室收缩功能明显强于对照组，并且心肌梗死范围最小。结论: MSCs移植后可以向心肌细胞分化，粒细胞集落刺激因子可以动员机体内自体MSCs到心肌缺血区域，两者共同应用对于缺血心肌有协同治疗作用。     

Haematological response of dogs to canine recombinant granulocyte colony stimulating factor (rcG-CSF)

Three beagle dogs were given 5 g canine recombinant granulocyte colony stimulating factor (rcG-CSF)/kg/day for 42 days. One day after the first dose the neutrophil count exceeded the pretreatment counts by 3- to 4-fold. A steady and rapid increase occurred during days 1 to 12. In two of the dogs the counts continued to increase, but at a slower rate, from day 14 to 28. The neutrophil count in the third dog decreased steadily from day 14 to 28, although the count remained above those of the presample period and exceeded the reference range for dogs established at the Veterinary Medical Teaching Hospital, University of California, Davis. After day 28 that dog had a rapid increase in neutrophil count that reached similar numbers to the other two dogs at 34 days. At no time was a significant left shift found, although an occasional band neutrophil was observed. In addition, a moderate increase in lymphocyte and monocyte counts were found. The degree of these increases was much less than that of the neutrophils. The leukocyte counts decreased rapidly after the last dose, and 10 days later the counts were similar to those of the pretrial period.Concomitant with the marked neutrophilia, bone marrows showed myeloid hyperplasia. The myeloid: erythroid ratios increased from 1.03–1.65 to 2.77–7.03, and the marrow cellularity increased from approximately 30%–50% to about 85%–100%. Clinical evaluation and serum chemistry panels revealed no adverse affects. We conclude that rcG-CSF effectively sustains an increased neutrophil count without producing significant adverse effects.     

巨噬细胞集落刺激因子及其受体mRNA在人卵泡颗粒细胞和胎盘及子宫内膜中的表达

目的： 观察巨噬细胞集落刺激因子（MCSF）及其受体（MCSFR）在人卵泡颗粒细胞、胎盘及子宫内膜细胞中的表达情况，进一步了解它们在生殖生理中的作用。方法： 收集人卵泡颗粒细胞、胎盘及子宫内膜细胞标本。采用逆转录-聚合酶链式反应（RT-PCR）及原位逆转录-聚合酶链式反应（in situ RT-PCR）技术观察MCSF及MCSFR mRNA的表达情况。结果： 在人卵泡颗粒细胞、胎盘及子宫内膜细胞中，RT-PCR显示有MCSF及MCSFR的表达，原位RT-PCR也进一步证实了这一点,并定位于细胞浆。结论： MCSF可能参与了人卵泡颗粒细胞、胎盘及子宫内膜细胞生长、发育的调节，在生殖生理过程中可能起一定的作用。     

重组人粒细胞巨噬细胞集落刺激因子对脓毒症模型小鼠的治疗作用

BACKGROUND: Granulocyte colony-stimulating factor has been shown to be used for the treatment of granulocytopenia, but its effect on sepsis is little reported. OBJECTIVE: To explore the treatment outcomes of recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF) for sepsis mice. METHODS: A sepsis mouse model was established by intraperitoneal injection of lipopolysaccharide, and then given subcutaneous injection of rhGM-CSF at 6 and 30 hours after modeling. The morphological change of mouse lung tissue, CD64 expression in peripheral neutrophils, and serum levels of tumor necrosis factor-α and interleukin-10 were ovserved, respectively. RESULTS AND CONCLUISON:(1) Complete bronchial epithelial, mild stromal hyperplasia, and a few neutrophil infiltration were found after rhGM-CSF treatment. (2) CD64 expression in peripheral neutrophils, and serum level of interleukin-10 in the treatment group were significantly higher than those in the model and control groups at 1, 3 and 7 days after treatment (P < 0.05). The serum level of tumor necrosis factor-α in the treatment group was significantly higher than that in the control group, but lower than that in the model group after 1, 3 and 7 days of treatment (P < 0.05). (4) These results show that rhGM-CSF can enhance neutrophil function and the anti-inflammatory effects in sepsis mice. 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程     

Neutrophil apoptosis: impact of granulocyte macrophage colony stimulating factor on cell survival and viability in chronic kidney disease and hemodialysis patients

Introduction

Altered neutrophil apoptosis might be responsible for recurrent bacterial infections encountered in hemodialysis (HD) and chronic kidney disease (CKD) patients. This work was designed to assess the neutrophil apoptotic activity and the impact of implementation of granulocyte macrophage colony stimulating factor (GM-CSF), as a survival factor, on neutrophil apoptosis among these patients.

Material and methods

Twenty-five patients on regular HD along with 34 CKD patients on conservative treatment, as well as 15 healthy controls, were investigated for apoptotic rate via assessment of neutrophil expression of Annexin-V by flow cytometry, before and after 20 h culture in absence and presence of GM-CSF. Neutrophil viability was determined using light microscopy. The preservation of neutrophil activation in these patients was analyzed by flow cytometric CD18 neutrophil expression. Chronic inflammatory state was evaluated by estimating C-reactive protein (CRP) and soluble intercellular adhesion molecule-1 (sICAM-1). Obtained data were statistically analyzed.

Results

Compared to controls, both HD and CKD groups had a significant increase of Annexin-V and CD18 expression and significant decrease in neutrophil viability. Culture of their neutrophils with GM-CSF showed significant decrease of apoptosis accompanied by improvement of neutrophil viability compared to their cultured cells without GM-CSF. These patients also showed significant elevation of CRP and sICAM-1.

Conclusions

Granulocyte macrophage colony stimulating factor demonstrated an evident impact on improving in vitro neutrophil survival and viability in HD and CKD patients. Therefore, this may represent promising preventive and/or therapeutic strategies against infection frequently observed in these patients and causing morbidity and mortality.     

粒细胞-巨噬细胞集落刺激因子治疗肺泡蛋白沉积症1例

收集1例肺泡蛋白沉积症(pulmonary alveolar proteinosis,PAP)患者临床资料并相关文献复习.患者因气短4个月,加重1个月入院,行纤维支气管镜活检.经过碘酸雪夫(Periodic acid-Schiff,PAS)染色阳性明确诊断为PAP,在经2次全肺灌洗治疗,病情反复后给予皮下注射粒细胞-巨噬细胞集落刺激因子,病情得到改善,提示对特发性PAP患者皮下注射粒细胞-巨噬细胞集落刺激因子是一种可行的治疗方法.     

粒细胞集落刺激因子促进大鼠梗死心肌修复

目的观察粒细胞集落刺激因子G(-CSF)对急性心肌梗死大鼠梗死区的修复和心功能的改善作用。方法29只大鼠随机分为细胞因子组(GAMI,n=9)、心肌梗死对照组(AMI,n=10)和假手术组(SO,n=10)。GAMI组和AMI组结扎左冠状动脉造成心肌梗死,SO组行相似的手术操作但不结扎冠状动脉。GAMI组给予rhG-CSF(50μg.kg-1.d-1),腹腔内注射,共7d。AMI组及SO组则每日腹腔内注射同等体积的生理盐水,共7d。心肌梗死后4周,通过心脏插管测量血流动力学参数,并在显微镜下观察梗死范围和梗死区、梗死周边区小血管数量的变化。结果AMI组大鼠有3只分别于术后第3、7、8天死亡,GAMI组与SO组大鼠至观察结束无一只死亡。与SO组相比较,AMI组LVSP(P<0.01)、dpd/tmax(P<0.01)、dpd/tmin(P<0.05)降低,LVEDP明显升高(P<0.01);GAMI组dpd/tmax(P<0.05)、LVEDP升高(P<0.01),LVSP、dpd/tmin无明显变化(P>0.05)。与AMI组相比较,GAMI组LVSP(P<0.01)、dpd/tmax(P<0.01)、dpd/tmin(P<0.05)升高,LVEDP降低(P<0.05)。与AMI组相比较,GAMI组梗死范围明显缩小(P<0.01),梗死区及梗死周边区小血管密度增加(P<0.05)。结论应用G-CSF可缩小心肌梗死大鼠梗死范围,提高梗死区及梗死周边区小血管密度并改善心功能。     

粒细胞集落刺激因子干预性治疗大鼠心肌梗塞的实验研究

目的：探讨用粒细胞集落刺激因子（G-CSF）干预性治疗心肌梗塞动物模型的作用。方法：健康大鼠随机分为心肌梗塞干预性治疗组和心肌梗塞对照组。干预性治疗组予G-CSF 10 μg·kg^-1·d^-1皮下注射5 d,并于第3 d用盐酸异丙肾上腺素（ISO）5 mg/kg腹腔注射复制大鼠心肌梗塞模型。对照组皮下注射等量生理盐水5 d,也于第3 d用ISO复制心肌梗塞模型。两组均于注射ISO后2周、4周将动物处死,取出心脏,病理检查大鼠心肌梗塞的病理变化情况。结果：注射ISO后2周、4周,干预性治疗组心室肌疤痕组织面积显著小于对照组（P<0.05）,并可见到心肌细胞、平滑肌细胞、血管内皮细胞的再生。结论：G-CSF能促进梗死后心肌组织再生,明显减少心肌梗塞后疤痕组织范围,可用于急性心肌梗塞的干预性治疗。