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.     

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     

Treatment with granulocyte colony stimulating factor is associated with improvement in endothelial function

Primary objective: Granulocyte-colony stimulating factor (G-CSF) is used for the mobilization of bone marrow and endothelial progenitor cells, though G-CSF-induced inflammation may cause endothelial dysfunction. We examined the effects of G-CSF on endothelium, C-reactive protein (CRP), tumour necrosis factor-α (TNF-α) and anti-inflammatory cytokines namely interleukin 10 (IL-10).

Research design: We studied 60 women with breast cancer, who were randomized to either subcutaneous G-CSF (5 μg/kg), o.d. for 5 days after adjuvant chemotherapy (n = 40) or placebo (n = 20).

Experimental interventions: We measured flow-mediated dilatation (FMD%) of the brachial artery by ultrasonography, CRP, TNF-α, IL-10 and the ratio TNF-α/ IL-10 blood levels before, 2-h and 5-days after the G-CSF or placebo treatment.

Main outcomes and results: There was a greater increase of FMD, IL-10 and reduction of TNF-α/ IL-10, 2 h and 5 days after the G-CSF treatment compared to placebo. Although, CRP and TNF-α were higher, TNF-α/IL-10 was lower at the end of G-CSF treatment compared to placebo. Improvement of FMD was related to changes of IL-10 and TNF-α/IL-10.

Conclusions: Treatment with G-CSF improves endothelial function in vivo, possibly by shifting the balance between the pro- and anti-inflammatory cytokines.     

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.     

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。     

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。     

Granulocyte-macrophage colony-stimulating factor (GM-CSF) but not granulocyte colony-stimulating factor (G-CSF) induces plasma membrane expression of proteinase 3 (PR3) on neutrophils in vitro

The theoretical risk of triggering vasculitis resulting from administration of G-CSF and GM-CSF to patients with anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitides (AAV), such as Wegener's granulomatosis (WG), who develop agranulocytosis due to cytotoxic therapy, is unknown. Since there is strong evidence that activation of polymorphonuclear neutrophils (PMN) induced by binding of ANCA to PR3 or myeloperoxidase (MPO) expressed on their plasma membrane is involved in the pathogenesis of systemic vasculitides (SV), we studied the surface expression of PR3 and MPO on PMN from healthy donors in response to G-CSF and GM-CSF in vitro by flow cytometric analysis. Increasing doses of G-CSF did not alter PR3 expression on either untreated or tumour necrosis factor-alpha (TNF-alpha)-primed donor PMN significantly. In contrast, GM-CSF significantly increased PR3 membrane expression on both intact PMN and neutrophils primed with TNF-alpha. MPO expression was not significantly altered by either G-CSF or GM-CSF. In summary, these data demonstrate that GM-CSF, but not G-CSF, induces plasma membrane expression of PR3 on PMN in vitro. Since in AAV accessibility of the antigen (PR3 or MPO) to the antibody (ANCA) on the plasma membrane of PMN is thought to be essential for neutrophil activation by ANCA, the results of the present study suggest that administration of GM-CSF to patients with WG with neutropenia implies a definite theoretical risk of deterioration of vasculitis via this mechanism.     

Hypopharyngeal squamous cell carcinoma producing both granulocyte colony-stimulating factor and parathyroid hormone-related protein

A 57-year-old woman was admitted to Hokkaido University Hospital because of dysphagia. Laryngoscopy indicated hypopharyngeal tumor histologically diagnosed as squamous cell carcinoma (SCC). A combination of radiotherapy and chemotherapy was performed for 2 months, and the hypopharyngeal lesion completely regressed. After 4 months, fever, anorexia, and malaise appeared, and chest X-ray and CT indicated two large tumors in the right lung. Transbroncheal lung biopsy (TBLB) specimens were diagnosed as SCC. Laboratory data showed high levels of serum granulocyte colony-stimulating factor (G-CSF) and parathyroid hormone-related protein (PTHrP). Subsequently, positron emission tomography (PET) showed multiple metastases to several organs including the liver, spine, skull, and thigh. Two months after readmission, the patient died with no success of chemotherapy. At autopsy, the lung tumor was clearly positive for both G-CSF and PTHrP on immunohistostaining. Retrospectively, examination showed that the primary pharyngeal tumor was focally positive for these two cytokines. Thus, a certain population of hypopharyngeal cancer producing G-CSF and PTHrP, spread to various organs and contributed to the rapid progression and poor prognosis. This case is presented with a discussion of several other cases in the literature.     

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.     

Granulocyte colony stimulation factor (G-CSF) suppresses interleukin (IL)-12 and/or IL-2 induced interferon (IFN)-gamma production and cytotoxicity of decidual mononuclear cells

PROBLEM: The placenta is one of the few non-hematopoietic tissues to express granulocyte colony stimulation factor (G-CSF). Placental G-CSF production is considered to be one of the major causes of granulocytosis during pregnancy although its physiological role in pregnancy has not yet been examined. METHOD OF STUDY: The effects of G-CSF on interleukin (IL)-2 and/or IL-12 induced interferon (IFN)-gamma production of magnetic cell sorting (MACS) sorted decidual lymphocytes was examined by enzyme-linked immunosorbent spot-forming cell assay (ELISPOT). The effect of G-CSF on cytotoxicity of decidual lymphocytes against the choriocarcinoma cell line JEG-3 was examined by lactate dehydrogenase (LDH) release assay. RESULTS: As previously reported by us, IL-2 and/or IL-12 activated decidual mononuclear cells were capable of killing choriocarcinoma cells. We observed that G-CSF abolished IFN-gamma production and cytotoxicity of decidual mononuclear cells and MACS sorted CD56+ cells. CONCLUSIONS: In addition to its well-known trophic effects on hematopoiesis, our results suggest about new roles of G-CSF in reproductive immunology.     

Granulocyte colony‐stimulating factor‐producing pancreatic anaplastic carcinoma in ascitic fluid at initial diagnosis: A case report

Granulocyte colony‐stimulating factor (G‐CSF)‐producing pancreatic tumors are extremely rare. These tumors have an aggressive clinical course and no established treatment. Here, we report an autopsy case of G‐CSF‐production in pancreatic anaplastic carcinoma (PAC). A 72‐year‐old woman presented with a large pancreatic head mass and multiple liver metastases. Laboratory data showed leukocytosis (leukocyte count 113.3 × 10³/µL) and high serum G‐CSF levels (441 pg/mL; normal range: <39.0 pg/mL). The ascitic fluid was submitted to our pathology laboratory at initial diagnosis. Cytopathology showed that smears from the ascitic fluid were highly cellular and contained numerous malignant cells, mainly in loose groupings. Occasional pseudoglandular formations and giant cells were also present. The malignant cells were round, and no spindle‐shaped cells were visible. The nuclei were round to ovoid with coarsely granular chromatin and large prominent nucleoli. Upon immunocytochemistry, tumor cells were positive for G‐CSF and vimentin; there was no E‐cadherin expression. Histopathological examination of the tumor showed a mixed composition of adenocarcinomatous and sarcomatous regions. Upon immunohistochemistry, both components were positive for G‐CSF. Few CD34‐positive myeloblasts were observed in the bone marrow. Thus, we diagnosed this as a case of G‐CSF production in PAC with leukocytosis. To the best of our knowledge, this is the first report on G‐CSF expression immunocytochemically confirmed in PAC. Diagn. Cytopathol. 2017;45:463–467. © 2017 Wiley Periodicals, Inc.     

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.     

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。     

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.     

Micropapillary component of urothelial carcinoma detected in transurethral resection of bladder tumor (TUR-BT) tissues: a case report

A case of urothelial carcinoma (UC) containing a micropapillary carcinoma (MPC) component in the urinary bladder of an 83-year-old man is reported. The MPC component of UC has been reported to be a variant featuring poor prognosis and rapid progression. In the present case, a characteristic MPC component with micropapillary growth, in association with a fine meshwork-like stroma, was observed in less than 10% of fragmented cancer tissues of UC, G3, obtained by transurethral resection of a bladder tumor (TUR-BT). Lymphatic invasion was also detected. UC cancer cells had invaded the prostatic glands and replaced the original epithelial cells. The unique "insideout" feature of the MPC component was immunohistochemically obvious on staining with antibody to epithelial membrane antigen (EMA). On immunohistochemical study, cancer cells of both UC and MPC components were positive for pancytokeratin AE1/AE3 and cytokeratins 7 and 20. Carcinoembryonic antigen (CEA) and CAM5.2 were only focally positive in UC cells. MIB-1(Ki-67) labeling index was high, at 80%-90%, in cancer cells of UC. This was a case of UC, G3 with invasion to the muscularis propria layer of the urinary bladder and also to the prostate. MPC and MPC components in cancers should be recognized as a marker of poor prognosis, even when detected in less than 10% of UC within TUR-BT tissues, as in the present case.     

Plasmacytoid urothelial carcinoma of the urinary bladder: A case report and immunohistochemical study

We report a rare case of plasmacytoid urothelial carcinoma (PUC) of the urinary bladder. A 50-year-old man complained of pollakiuria and urinary incontinence. MRI detected a bladder tumor invading the rectum and bilateral hydroureteronephrosis. Radical cystectomy with partial resection of the rectum was performed, and ileus due to peritoneal dissemination occurred 2 years after surgery. He died of the disease 42 months after the initial presentation. Histologically, urothelial carcinoma in situ with a focal invasive urothelial carcinoma (IUC) component and widely spread PUC was observed. There was no lymph node metastasis. PUC cells had eccentrically placed nuclei and eosinophilic cytoplasm resembling plasmacytoma cells, and proliferated with a single-cell infiltrative pattern to the outside of the bladder. IUC cells with intracytoplasmic lumina were focally intermingled with PUC cells. Immunohistochemically, PUC cells were positive for cytokeratin 7, epithelial membrane antigen, and CA19-9, but negative for cytokeratin 20, E-cadherin, p63, and lymphoid markers. The Ki-67 labeling index of PUC cells was 9.3%. IUC containing intracytoplasmic lumina showed intermediate features of conventional IUC and PUC morphologically and immunohistochemically. PUC is a distinct entity of bladder cancer with a high propensity for invasion and poor prognosis.     

Primary urinary bladder adenosquamous carcinoma complicated with lower limb deep venous thromboses: a case report

Primary urinary bladder adenosquamous carcinoma is extremely rare and only a few cases have been reported in English literatures. Its biological behavior remains unclear. Here we reported a 60-year-old male patient with lower limb deep venous thromboses associated with primary urinary bladder adenosquamous carcinoma. A color ultrasonography showed right stock total venous thrombosis and right great saphenous vein thrombosis of lower limb. Contrast-enhanced computed tomography (CT) scan confirmed a 3.17 × 3.33 × 3.84 cm enhancing mass within the urinary bladder along the right lateral and posterior wall. Histopathological examination revealed adenosquamous carcinoma of urinary bladder, with extensive infiltration of the muscle layer. To the best of our knowledge, this is the first report of primary urinary bladder adenosquamous carcinoma complicated with deep venous thromboses in lower limb.     

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.     

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.