Granulocyte colony stimulating factor treatment for alloimmune neonatal neutropenia.

Granulocyte colony stimulating factor (G-CSF) treatment was successfully used in three preterm infants with alloimmune neonatal neutropenia (AINN). Two infants had persistent neutropenia despite treatment with intravenous immunoglobulin and random donor granulocyte transfusions for presumed sepsis. Neutrophil counts returned to normal with G-CSF treatment; the response was least convincing in one infant with fulminant necrotising enterocolits. It is suggested that treatment with G-CSF be considered early for the treatment of infants with AINN.     

Granulocyte colony stimulating factor in neonatal alloimmune neutropenia: A possible association with induced thrombocytopenia

Neonatal alloimmune neutropenia (NAIN) is a rare cause of congenital neutropenia seen in <1% of births. Significant morbidity, usually infections, may result from this disease. The pathophysiology of NAIN, mediated by maternal antibodies crossing the placenta to destroy fetal cells expressing paternal antigens, is similar to that of neonatal alloimmune thrombocytopenia, as well as Rh/ABO hemolytic disease of the newborn. The use of high‐dose granulocyte colony stimulating factor in patients with NAIN may cause a reversible thrombocytopenia in some patients. Pediatr Blood Cancer 2010;54:1014–1016 © 2010 Wiley‐Liss, Inc.     

The in vivo effect of recombinant human granulocyte-colony stimulating factor in neutropenic neonates with sepsis

The effects of recombinant granulocyte-colony stimulating factor (rhG-CSF) in neonatal neutropenia with presumed sepsis, which has a poor prognosis, were investigated. The study involved 14 neonates with presumed sepsis and neutropenia. Findings were compared with those from 24 historical controls. rhG-CSF (5 μg/kg/day i.v. for 5 days) was administered immediately following diagnosis. Complete blood counts were obtained before and 24, 48, 72, 96 and 120 h after initiation of treatment. Neutrophil storage pool (NSP) was assessed (in 4 patients) before and after treatment. Statistical analysis was performed using one way analysis of variance. Treatment led to an increase in absolute neutrophil count (ANC) levels in 13/14 patients. At the end of treatment, the mean ANC was higher than that of controls (P = 0.007). There was a marked increase in the NSP of between 32% and 65% (P = 0.005). There were two clinical failures, one of whom was considered to have died from his underlying condition. There were no reports of clinical or haematological toxicity during treatment or follow up. Received: 14 June 1996 / Received in revised form and accepted: 15 November 1996     

儿童中性粒细胞减少症研究进展

中性粒细胞是参与宿主抗感染过程的一种重要的免疫细胞。中性粒细胞减少症是表现为外周血循环中成熟中性粒细胞绝对值减少的一组疾病,通常伴随细菌感染风险的增加。根据病因及发病机制可分为先天性和后天获得性。本文综述近期儿童中性粒细胞减少症病因学研究现状及进展。深入了解中性粒细胞减少症的病因,有助于提高中性粒细胞减少症的诊治水平。     

Granulocyte colony-stimulating factor receptor expression on neutrophils of term and preterm neonates with and without signs of infection

Neutrophils are an essential component of the human host defence system against infection. Recombinant human granulocyte colony-stimulating factor induces neutrophilia and enhances effector functions of mature neutrophils. Since the biological effects of granulocyte colony-stimulating factor (G-CSF) are mediated by its receptor, we investigated the expression of G-CSF receptor on the surface of neutrophils of term and preterm neonates (n = 22) with and without signs of infection and of healthy adults (n = 13) by flow cytometry. In healthy adults, the percentage of neutrophils expressing G-CSF receptor was higher compared to cord blood of term and preterm neonates (87% vs 53%, P < 0.05). Between 2 and 32 h of life, neonates with signs of infection showed lower values of G-CSF receptor expression compared to neonates without signs of infection (32% vs 54%, P < 0.05). No correlation was detectable between expression of G-CSF receptor and gestational age. Conclusion Expression of granulocyte colony-stimulating factor receptor on neutrophils is lower than in adults. This may adversely affect granulopoiesis and neutrophil function during the neonatal period. Moreover, granulocyte colony-stimulating factor receptor expression seems to be down-regulated during neonatal infection. Received: 8 June 1998 / Accepted in revised form: 13 October 1998     

Improvement of neutropenia and neutrophil dysfunction by granulocyte colony-stimulating factor in a patient with glycogen storage disease type Ib

Patients with glycogen storage disease type Ib (GSD Ib) suffer from recurrent bacterial infections due to neutropenia and neutrophil dysfunction. To improve the quality of life in a 9-year-old boy with GSD Ib, we subeutaneously administered recombinant human granulocyte colony-stimulating factor (G-CSF). Daily injections of 100 g/m² of G-CSF significantly increased absolute neutrophil counts and augmented neutrophil mobility. The patient was then treated with 70 and 100 g/m² of G-CSF daily and twice-weekly. The treatment maintained absolute neutrophil counts at significantly higher levels than those without treatment for 22 months and markedly decreased the frequency of infections and the necessity for hospitalisation. No adverse effects were observed during treatment. These findings indicate that daily and twice-weekly treatment with G-CSF of long duration are safe and effective for patients with GSD Ib. G-CSF may be a useful therapeutic agent in patients with neutrophilic impairment as a consequence of a metabolic disorder.     

Differential diagnosis and clinical course of autoimmune neutropenia in infancy: comparison with congenital neutropenia

Aim: Autoimmune neutropenia in infancy (AIN) is caused by granulocyte-specific autoantibodies. Clinical presentation and diagnosis have not been well studied, resulting in cumbersome diagnostic investigations and unnecessary treatment such as granulocyte colony-stimulating factor (G-CSF) therapy. Methods: Clinical, laboratory and immunological data of 18 infants with AIN were evaluated. Granulocyte-specific autoantibodies were detected by the direct granulocyte immunofluorescence test (D-GIFT), indirect granulocyte immunofluorescence test (I-GIFT) and immunoblotting. Results: The average age of onset and resolution of neutropenia in AIN was 7.4 ±3.4 mo (mean ±SD) and 20.4 ±4.9 mo, respectively. Sixteen of the 18 patients presented with mild infectious symptoms; the other 2 patients were detected by chance and presented with no infectious symptoms. D-GIFT was positive in all patients, and I-GIFT was positive in 17 of these 18 patients. Most patients showed preferential binding to neutrophils from NA(1 + 2-)-phenotyped donors by I-GIFT and immunoblotting. An antibiotic (sulfamethoxazole-trimethoprim) was given to 15 patients for prophylaxis. G-CSF was given to only one infectious patient.

Conclusion: A combination of diagnostic tests for the detection of granulocyte-specific autoantibodies was useful in diagnosing AIN, thus avoiding unnecessary investigations. Continuous treatment with G-CSF was not necessary for prophylaxis, even if neutrophil counts were extremely low.     

Granulocyte colony-stimulating factor in glycogen storage disease type 1b. Results of the European study on glycogen storage disease type 1

Patients with glycogen storage disease type 1b (GSD-1b) have neutropenia and neutrophil dysfunction that predispose to frequent infections and inflammatory bowel disease (IBD), for which granulocyte colonystimulating factor (GCSF) is given. To investigate the use and the value of GCSF treatment in GSD-1b, a retrospective registry of GSD-1 patients born between 1960 and 1995 in 12 European countries was established. Included were 57 GSD-1b patients. Unglycosylated GCSF was given to 18 patients, median age of starting therapy was 8 years, longest duration of therapy 7 years. Dose varied between 2–10 μg/kg, with a frequency from daily to twice per week. Neutropenia (defined as an absolute neutrophil count <0.5×10⁹/1) was found in 49 patients. In untreated patients, a significant decrease of haemoglobin, platelet counts and leucocyte counts with increasing age (P<0.032,P<0.04 andP<0.001 respectively) was noted, whereas neutrophil counts remained low but stable with increasing age. In nine patients who were treated longer than 1 year, median neutrophil counts increased significantly and simultaneously median leucocyte counts and platelet counts decreased significantly. In all patients treated, the number and severity of infections decreased and the severity of IBD improved subjectively. The most serious complication of GCSF treatment was marked splenomegaly (four patients).Conclusion: in this retrospective study a significant haematological effect was documented and a subjective improvement of infections and inflammatory bowel disease. In view of the uncertainty, prospective controlled trials seem warranted to clarify the indication for the use of granulocyte colony-stimulating factor in this disease. Published online: 17 July 2002     

GM-CSF联合乙肝疫苗治疗宫内HBV感染慢性携带儿童的研究

目的 初步观察重组粒细胞－巨噬细胞集落刺激因子（GM－CSF）或乙肝免疫球蛋白（HBIG）联合酵母重组乙型肝炎疫苗（rHBvac）对宫内感染呈慢性乙型肝炎病毒（HBV）携带儿童的治疗作用。方法 母亲为慢性HBV携带者，出生时乙肝表面抗原(HBsAg）阳性经乙型肝炎疫苗免疫仍发展为慢性HBV无症状携带儿童27名，随机分为两组。Ⅰ组：14名，使用GM－CSF50μg和rHBvac 10μg同一部位肌肉注射，每月1次，共4次。Ⅱ组：13名，HBIG 200IU和rHBvac10μg不同部位肌肉注射，每月1次，共4次。治疗结束后1个月复查HBV－DNA定量及其他HBV标记。结果 两组各有12名儿童完成研究，治疗前丙氮酸转氨酶异常Ⅰ组3例，Ⅱ组4例，治疗后各有2例恢复正常；治疗前乙型肝炎e抗原（HBeAg）阳性Ⅰ组9例，Ⅱ组10例，治疗后各1例发生HBeAg/抗－HBe血清转换。HBV－DNA定量，治疗后和治疗前相比，Ⅰ组有显著下降（符号铁和检验P＝0.023），Ⅱ组变化不明显，两组中无HBsAg转阴病例。结论 GM－CSF联合乙肝疫苗对乙肝疫苗免疫失败的慢性HBV携带儿童有一定治疗作用。     

中性粒细胞缺乏合并脓毒症时前B细胞克隆增强因子水平的研究

目的研究前B细胞克隆增强因子(PBEF)在中性粒细胞缺乏状态合并脓毒症时的水平变化及临床应用价值。方法选取28例急性淋巴细胞白血病(ALL)化疗后处于中性粒细胞缺乏状态的患儿为研究对象,其中无感染14例,脓毒症14例。收集所有患儿的临床资料及血清标本,用酶联免疫吸附法(ELISA)检测PBEF浓度,比较两组PBEF水平,分析PBEF与临床常用感染指标CRP、PCT的相关性,探讨中性粒细胞缺乏时PBEF在脓毒症的临床应用价值。结果脓毒症组PBEF浓度为(5.526±1.548)ng/mL,无感染组PBEF浓度为(5.184±1.295)ng/mL,两组PBEF水平比较无明显差异(p=0.536)。脓毒症组PBEF与其他常见炎性指标CRP、PCT无显著相关(尸值分别为0.159和0.311)。结论中性粒细胞缺乏状态合并脓毒症时PBEF水平无明显改变,可能与外周血中合成和分泌PBEF的细胞减少相关,PBEF是否可作为感染指标应用于临床有待进一步探讨。     

Effect of granulocyte and granulocyte macrophage colony stimulating factors (G-CSF and GM-CSF) on neonatal neutrophil functions

Although there are many studies on the effect of granulocyte and granulocyte-macrophage colony stimulating factors (G/GM-CSF) on adult neutrophil functions, there is little information regarding their influence on neonatal cells. We studied the in vitro effect of G/GM-CSF on neutrophil chemotaxis, polarization, and superoxide anion generation in 47 neonates compared with 35 adults. We found that G-CSF and GM-CSF significantly enhanced the chemotaxis of newborn infants' neutrophils, normalizing their chemotactic defect [from 35 +/- 7 cells/field (mean +/- SE) to 49 +/- 5 cells/field with G-CSF, p < 0.05 and to 55 +/- 4 cells/field with GM-CSF, p < 0.001]. It is notable that the maximal neutrophil response to the cytokines was observed particularly in the newborn infants with severe impairment in their chemotactic activity. Statistical analysis of the data showed a significant inverse correlation, which supported this observation (r = -0.6, p < 0.02 for G-CSF; r = -0.76, p < 0.001 for GM-CSF). The reduced polarization of neonatal compared with adult cells [71 +/- 5% versus 86 +/- 2% (mean +/- SE), p < 0.05], was corrected by CSF-priming (to 87 +/- 4% with G-CSF and to 92 +/- 2% with GM-CSF, p < 0.05). In addition, the neutrophil superoxide generation was significantly improved in both groups following the CSF-priming. GM-CSF and G-CSF gave comparable results in all functions studied except that GM-CSF improved superoxide release to a greater extent. This study shows a significant improvement of the neonatal neutrophil functions following in vitro CSF-priming and contributes to a better understanding of the neonatal neutrophil behavior when treated with G/GM-CSF.     

Protective effects of recombinant human granulocyte colony stimulating factor in a rat model of necrotizing enterocolitis

The role of cytokines and growth factors in the pathophysiology of neonatal necrotizing enterocolitis (NEC) is not defined clearly yet. The aim of this study was to determine the effects of recombinant human granulocyte colony stimulating factor (G-CSF) on intestinal cells in hypoxia-induced experimental NEC in rats. The study was experimented on Sprague Dawley rat pups. Group 1 (untreated, n = 7) rats were subjected to hypoxia–reoxygenation (H/O) and then were returned to standard conditions. Group 2 (G-CSF treated, n = 7) rats were subjected to H/O, and then were treated with G-CSF (100 μg/kg enterally) for 5 days. Group 3 was served as nonhypoxic controls. All animals were killed on day five, and histological examination was performed on intestinal samples. There were no histopathological changes in the control group. The histological findings in untreated rats were similar to those seen in neonatal NEC, with destruction of villi and crypts with extension to the muscularis layer. Intestinal damage was mild in group 2 and these histological changes were better than group 1, and worse than group 3. The mean of histologic grade of group 1 was 2.4 (range 2–3), and in the group 2, it was 1.2 (range 0–2). A difference was found when two groups were compared with each other (P < 0.05). In an experimental model of NEC, G-CSF could have a protective effect on intestinal damage.     

Relationships of cytokine (GM-CSF) serum concentration to blood cell count and the inflammatory parameters in children with malignant diseases

This study examined both the endogenous and exogenous (therapy-related) pharmacokinetics of the cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) in neutropenic children with solid and systemic malignancies. The daily endogenous GM-CSF serum concentration before application was 29 pg/mL. During the 10 days of examination, the concentration rose to an average value of 1351 pg/mL 8 h after application. A significant stimulation of the neutrophils, monocytes, and eosinophils in peripheral blood was documented. No significant correlation between the GM-CSF concentration and peripheral blood cell counts could be documented. Paraclinically, there was no evidence for functional disturbance of either the liver or the kidneys, i.e., under the cytokine therapy. The therapy was well tolerated by all the children involved in the study.     

Plasma erythropoiesis stimulating factor(s) in neonatal mice: in vitro dose response and chromatography studies

High levels of plasma erythropoiesis stimulating factor(s) (ESF) have been found in neonatal WLO-mice during rapid growth. A previous study on hypertransfused neonatal animals indicated that the high ESF could not be due to the concomitant postnatal anemia alone. The present investigation was performed to answer the question: Is the high plasma ESF in neonatal WLO-mice erythropoietin (Ep) alone, or Ep in combination with other factors? The ESF of plasma from 20-day-old animals and standard Ep were compared in a cell culture assay for ESF based on erythroid colony formation, and also by means of gel filtration chromatography and affinity chromatography. Nonfractionated plasma and standard Ep showed parallel dose response curves and additive activity in the ESF assay. After gel filtration the detectable ESF of plasma was eluted in the same position as that of standard Ep, corresponding to an estimated molecular weight range of 34-65,000 daltons. The ESF of intact plasma, fractionated plasma, and standard Ep were identically bound to and eluted from the affinity chromatography column. These results show that the ESF of plasma from 20-day-old animals can neither be separated into several factors, nor distinguished from that of standard Ep by the methods used. It is therefore concluded that the high plasma ESF found in neonatal WLO-mice probably consists of Ep alone.     

Granulocyte-colony stimulating factor levels in cord blood and neonatal peripheral blood

We measured the granulocyte-colony stimulating factor (G-CSF) levels in cord blood and peripheral blood obtained from full-term or pre-term infants during the first 3 days of birth. The mean G-CSF level among cord blood (17.2pg/mL) was similar to that of peripheral blood on day 0 (18.3 pg/mL) and day 1 (13.6 pg/mL), while that of peripheral blood on day 0 was significantly higher than on day 2 (10.9 pg/mL) and day 3 (8.8 pg/mL; both P < 0.05). There was no correlation between neutrophil counts and G-CSF levels. No difference was found in neutrophil counts or G-CSF levels between infants who weighed more or less than 2500 g at birth. These results suggest that the neonatal neutrophil count depends on regulatory factors other than G-CSF.     

心肌疾病抗ADP/ATP载体抗体与粒细胞-巨噬细胞集落刺激因子的临床研究

目的探讨病毒性心肌炎（ＶＭＣ）和扩张型心肌病（ＤＣＭ）的自身免疫发病机制。方法采用免疫印转和放射免疫技术检测了３０例ＶＭＣ、１４例ＤＣＭ患儿血浆中抗心肌线粒体ＡＤＰ／ＡＴＰ运载蛋白（ＡＮＴ）抗体和粒细胞－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）。结果ＶＭＣ和ＤＣＭ患儿抗ＡＮＴ抗体和ＧＭ－ＣＳＦ的阳性率分别为７３％和５７％，而２５例正常健康儿分别为０和１２％（Ｐ均＜０．０１），且抗ＡＮＴ抗体与ＧＭ－ＣＳＦ的血浆水平呈正相关（ｒ＝０．４０９１，Ｐ＜０．０５）。结论小儿ＶＭＣ和ＤＣＭ的发病与心肌的自身免疫损伤有关，抗ＡＮＴ抗体和ＧＭ－ＣＳＦ均参与了这一过程。抗ＡＮＴ抗体可作为小儿ＶＭＣ和ＤＣＭ的一种特异性诊断指标。