Granulocyte Colony-stimulating Factor Preparations Research Articles

Nested Case-Control Study Utilizing MID-NET® on Thrombocytopenia Associated With Pegfilgrastim in Patients Treated With Antineoplastic Agents.

Although several spontaneous case reports on the occurrence of thrombocytopenia in patients treated with human granulocyte colony‐stimulating factor (G‐CSF) preparations have been accumulated, its actual causality is still unclear. To investigate the association between G‐CSF preparations (filgrastim, nartograstim, lenograstim, and pegfilgrastim) available in Japan and thrombocytopenia in patients treated with antineoplastic agents, a nested case‐control study was conducted using the Medical Information Database NETwork (MID‐NET®) with the cohort of the Japanese population taking antineoplastic agents between 2009 and 2018. A case of thrombocytopenia was defined as a patient who had decreased platelet counts (< 50,000/mm3). We identified a maximum of 10 controls for each case matched on the index date. Adjusted odds ratios (aORs) and their 95% confidence intervals (CIs) of thrombocytopenia for the use of G‐CSF preparations compared with nonuse were estimated using conditional logistic regression. From the cohort in which 33,124 patients were included, 733 cases and 5,592 controls were identified. Compared with the nonuse of G‐CSF preparations, the use of any G‐CSF preparations increased the risk of thrombocytopenia (aOR: 5.7, 95% CI: 4.3‐7.5). More detailed analysis showed that a distinctive increased risk was observed when pegfilgrastim was prescribed at 2–7 days before the index date (aOR: 7.4 95% CI: 2.0–28.1). Associations of the other G‐CSF preparations with thrombocytopenia were unclear due to the inconsistent results among different analyses. A significantly increased risk of thrombocytopenia associated with pegfilgrastim was identified, leading to a revision of precautions in the package inserts of pegfilgrastim as a regulatory safety action.

Factors for the optimal selection of granulocyte colony-stimulating factor preparations and predictors for R-CHOP dose reductions/delays among patients with non-Hodgkin B-cell lymphoma (STOP FN in NHL 2 subanalysis)

BackgroundA classification tree was used to analyze background factors for granulocyte colony-stimulating factor (G-CSF) preparation selection for febrile neutropenia (FN) prophylaxis in Japanese patients with non-Hodgkin B-cell lymphoma receiving the first R-CHOP cycle.MethodsThis was a subanalysis of the retrospective observational study STOP FN in NHL 2 (UMIN000029534). Patient characteristics, changes in neutrophil count, incidence and severity of neutropenia, and risk factors for dose reduction/delay of R-CHOP were assessed by G-CSF formulation.ResultsAmong 234 patients in cycle 1, 25.6% received no G-CSF preparation, 52.1% received daily G-CSF, and 22.2% received pegfilgrastim. Pegfilgrastim use was most frequent among patients aged ≥ 80 years, while that of daily G-CSF was most frequent in patients with lymphocyte count (LC) < 1000 cells/μL. Changes in neutrophil count were more marked with pegfilgrastim compared with daily G-CSF and no G-CSF. Relevant factors for G-CSF preparation selection in the first R-CHOP cycle were age ≥ 80 years and LC < 1000 cells/μL; for chemotherapy dose reduction were FN onset in cycle 1 and female sex; and for dose delay was hemoglobin (< 12 g/dL). After cycle 2 and onward, pegfilgrastim use increased markedly (72.6%) compared with cycle 1 (22.2%), with significantly greater proportions continuing pegfilgrastim use and switching from daily G-CSF.ConclusionRelevant factors for G-CSF preparation selection were age ≥ 80 years and LC < 1000 cells/μL. The use of pegfilgrastim increased markedly after cycle 2. These results may be useful for selecting appropriate G-CSF preparations in the first R-CHOP cycle.Trial registrationUMIN000029534; registered on 13 October 2017, https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000033733.

Use of pegylated granulocyte colony-stimulating factor in dose-adjusted EPOCH-R therapy

Dose-adjusted (DA)-EPOCH-R causes profound neutropenia requiring relatively long hospital stays with multiple doses of granulocyte colony-stimulating factor (G-CSF). A single-dose pegylated G-CSF (PEG-G-CSF) has been used for the treatment of chemotherapy-induced neutropenia. We retrospectively examined 15 patients (median age 61, range 33-75 years) treated with DA-EPOCH-R. In the first cycle of the DA-EPOCH-R therapy, a G-CSF preparation was used, and since the second cycle, the G-CSF and PEG-G-CSF use groups were divided. The median length of hospitalization after starting chemotherapy in the second-cycle DA-EPOCH-R was significantly shorter with PEG-G-CSF group (n=9) of 9 (7-13) days compared with G-CSF group (n=6) of 18 (15-22) days (P<0.001). Risk factors of febrile neutropenia, such as bone marrow invasion, performance status, serum albumin, and history of febrile neutropenia at the first DA-EPOCH-R cycle or previous chemotherapy were not significantly different for both groups, and the incidence of febrile neutropenia in PEG-G-CSF and G-CSF groups was 2.6% and 46.9%, respectively. These analyses suggest that PEG-G-CSF can be combined with DA-EPOCH-R without compromising treatment outcomes as compared with the daily dose of G-CSF.

Enhancing granulocyte colony-stimulating factor expression in Pichia pastoris through fusion with human serum albumin

ABSTRACTProtein fusion technology has emerged as one of the important strategies to increase the level of expression and half-life of therapeutic proteins in heterologous expression systems. Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic growth factor and is clinically used against neutropenia. Enhanced expression and stability of G-CSF were achieved in Pichia pastoris by the way of constructing a fusion protein with human serum albumin (HSA). The strategy involved polymerase chain reaction (PCR) amplification of fragments corresponding to codon-optimized G-CSF and domain 3 of HSA. Overlapping PCR was used to obtain the full-length fused gene (1,184 bp) with a 15-bp linker sequence comprising of 4 Gly and 1 Ser residues. Extracellular expression was carried out downstream of α-factor secretion signal sequence under the control of alcohol oxidase 1 promoter using pPICZαB. Excreted protein in the range of 110–380 mg L−1 was observed among the transformants. Effect of aeration and temperature was investigated in one of the transformants (35) overexpressing fusion protein and levels of G-CSF enhanced by 1.8-fold and 2.3-fold, respectively. Assay of biological activity indicated the fusion protein to retain similar cell proliferation activity as the commercial G-CSF preparation.

Hematologic Recovery in Patients Who Are Treated With Autologous Stem Cells Transplantation Taken From Bone Marrow After Granulocyte–Colony-Stimulating Factor Stimulation

Background We sought to compare hematologic recovery between patients who did or did not receive granulocyte–colony-stimulating factor (G-CSF)-stimulated bone marrow (rich bone marrow [RBM]). Materials and Methods The study subjects were 20 patients whose bone marrow was taken without prior stimulation with G-CSF and 15 patients in whom bone marrow was taken after previous G-CSF mobilization. The bone marrow harvest took place on the fifth day after G-CSF initiation. The bone marrow aliquot was 20 mL/kg. Results The median value of nucleated cells obtained from patients without G-CSF preparation was 3.65 × 10 8/kg. The median value of nucleated cells from RBM patients was 4.83 × 10 8/kg. The median value of stem cells obtained from patients without G-CSF preparation was 0.96 × 10 6/kg versus 1.9 × 10 6/kg from RBM patients. The median time to recovery of the hematopoietic system based on an increase in PLT value >20 g/L was 12.6 days for RBM versus 18.8 days without G-CSF preparation. The median time to recovery of the hematopoietic system based on assessment of growth ANC>0.5 g/L was 13.0 days for RBM versus 17.8 days without G-CSF stimulation. Significantly higher values of nucleated cells and increased stem cells were observed among RBM patients compared with those whose bone marrow was harvested without any stimulation ( P = .01). There was faster recovery of the hematopoietic system in cases where bone marrow was collected after G-CSF: PLT >20 g/L ( P = .015) and ANC >0.5 g/L ( P = .01). We also observed that the use of stimulated bone marrow shortened hospital stay after the administration of hematopoietic cells to 17.3 days compared with 23.1 days among patients receiving hematopoietic cells from nonstimulated bone marrow. The number of complications during transplantation was comparable in both cases, the most frequent ones being febrile neutropenia and grade III and IV mucositis. Conclusion RBM is a better method to obtain stem cells from bone marrow. Stimulated bone marrow shows faster engraftment compared with nonstimulated bone marrow helping patients who fail to generate are adequate number of stem cells from peripheral blood.

Superior mobilisation of haematopoietic progenitor cells with glycosylated G‐CSF in male but not female unrelated stem cell donors

Granulocyte colony-stimulating factor (G-CSF) effectively mobilises haematopoietic stem cells to the peripheral blood. It is unclear whether the mobilisation of stem cells with lenograstim (glycosylated G-CSF) or filgrastim (non-glycosylated G-CSF) leads to a higher cell number of collected engraft able progenitor cells. Thus, we investigated harvesting efficiency of the licensed G-CSF preparations in mobilising peripheral stem cells in a randomised study. A total of 501 healthy unrelated donors, including 339 males and 162 females received either lenograstim (n = 261) or filgrastim (n = 240) at 10 microg/kg body weight (BW) per day. Aphaeresis was performed on day 5 and, if necessary, on day 6 of mobilisation. The number of CD34+ cells collected was 11.5% higher in the lenograstim group (7.19 x 10(6) vs. 6.44 x 10(6)/kg BW donor; P < 0.03). Univariate variance analysis revealed that this effect was caused by male donors: more progenitors cells per kg BW of the donor (7.73 x 10(6) vs. 6.88 x 10(6); P < 0.017) and of the recipient (10.1 x 10(6) vs. 8.88 x 10(6), P < 0.029) could be harvested. There was no significant difference in the percentage of donors in whom a second aphaeresis was required (9.6% vs. 11.6%). Lenograstim mobilises progenitor cells into the peripheral blood more effectively in males than filgrastim.

Are Granulocyte Colony–Stimulating Factors Beneficial in Treating Diabetic Foot Infections?

To assess the value of granulocyte colony-stimulating factor (G-CSF) as adjunctive therapy for diabetic foot infections. We systematically searched the medical literature (including Medline, Embase, LookSmart, and the Cochrane Library) for prospective randomized studies that used G-CSF as an adjunct to standard treatment for diabetic foot infections. Using a conventional meta-analysis, we pooled the relative risks (RRs) for outcomes of interest, including resolution of infection, wound healing, duration of antibiotic therapy, and need for various surgical interventions, using a fixed-effects model. Five randomized trials, with a total of 167 patients, met our inclusion criteria. The methodological quality of the studies was satisfactory. The investigators administered various G-CSF preparations parenterally for between 3 and 21 days. The meta-analysis revealed that adding G-CSF did not significantly affect the resolution of infection or the healing of the wounds but was associated with a significantly reduced likelihood of lower extremity surgical interventions (RR 0.38 [95% CI 0.20-0.69], number of patients who needed to be treated: 4.5), including amputation (0.41 [0.17-0.95], number of patients who needed to be treated: 8.6). There was no evidence of heterogeneity among the studies or of publication bias, suggesting that these conclusions are reasonably generalizable and robust. Adjunctive G-CSF treatment does not appear to hasten the clinical resolution of diabetic foot infection or ulceration but is associated with a reduced rate of amputation and other surgical procedures. The small number of patients who needed to be treated to gain these benefits suggests that using G-CSF should be considered, especially in patients with limb-threatening infections.

Discrimination of granulocyte colony-stimulating factor isoforms by high-performance capillary electrophoresis

Granulocyte colony-stimulating factor (G-CSF) is a glycoprotein which acts primarily to stimulate the proliferation, differentiation and activation of committed progenitor cells of the neutrophil–granulocyte lineage into functionally mature neutrophils. The traditional biological assays employed to detect G-CSF are a myeloid bone marrow colony assay, a factor-dependent cell line specific for G-CSF and commercially available immunoassays. However, these methods will not distinguish between glycosylated and non-glycosylated forms of the molecule. In this study high-performance capillary electrophoresis (HPCE) was used to analyse glycosylated and non-glycosylated recombinant human granulocyte colony-stimulating factor (r-met-hG-CSF). Glycosylated G-CSF preparations contained human serum albumin (HSA), added as a protein carrier. Glycosylated and non-glycosylated G-CSFs were prepared in 40 m M Na 2HPO 4 buffer, pH 2.5, containing hydroxypropylmethylcellulose (HPMC) or 50 m M Na 2HPO 4 buffer, pH 9.0. Glycosylated G-CSF could be separated into two distinct glycoform populations at the lower pH studied. Differences in migration time and peak shape between glycosylated and non-glycosylated G-CSF were demonstrated. HPCE analysis of G-CSF produced using a baculovirus expression vector system revealed a further distinct G-CSF glycoform and demonstrated the resolving power of the technique.

Renal and Hematological Complications in Long-Term Treatment with Granulocyte Colony-Stimulating Factor 779

The value of granulocyte colony-stimulating factor (G-CSF) in the treatment of conditions associated with bone marrow aplasia/suppression is well appreciated. However, accruing experience is bringing to light adverse side effects that appear to be associated with long-term administration of G-CSF. We report 2 siblings with severe congenital neutropenia (SCN) (Kostmann's disease) who featured G-CSF (Filgrastim) -induced major renal and hematological complications. FA, a 12yr old girl and AA, her 17yr old brother, received daily G-CSF therapy for the last 9yrs. Both patients developed persistent microscopic hematuria with normal kidney function 2 mo after starting G-CSF. However, 4 yrs later, FA showed proteinuria, macroscopic hematuria and reduced kidney function. Renal biopsy revealed membranoproliferative glomerulonephritis type I. Discontinuation of G-CSF therapy for 4 months resulted in improvement of renal function. Persistent severe neutropenia and bacterial infections required resumption of G-CSF, which has led to deterioration of renal function and partially steroid-responsive hematuria and proteinuria. AA, who continues to show microscopic hematuria, has lately developed pancytopenia, with positive Coombs test and antiplatelet antibodies. Serology for parvovirus B19 was negative. Bone-marrow aspirates revealed maturation arrest of neutrophil precursors at the promyelocyte/myelocyte stage, and also megaloblastic changes and diminution of red cell progenitors and of megakaryocytes. Myeloid dysplasia or acute myeloid leukemia were not found. Each attempt to increase G-CSF dosage failed to raise the neutrophil count but aggravated the pancytopenia, requiring blood transfusions. In contrast, discontinuation of G-CSF resulted each time in a prompt reversal of pancytopenia and in disappearance of the Coombs and antiplatelet antibodies. AA was scheduled for bone marrow transplant and received during 33 days a glycosylated G-CSF (Lenograstim) preparation without adverse hematological phenomena.

Ｇ−ＣＳＦ製剤の持続静注における輸液セットへの吸着

The change in the adsorption to infusion sets, the adsorption ratio and the recovery ratio of recombinant human granulocyte-colony stimulating factor (G-CSF) were investigated in vitro by administering a continuous intravenous infusion of G-CSF.After 250μg of G-CSF (nartograstim) preparation was dissolved with 100 ml of sterile saline in a commercially produced polypropylene container, operating the pump (8 ml per hrs), the G-CSF levels in the container, in the upper part of the final filter, in the lower part of the final filter, and in the collecting vessels, were determined by enzyme-linked immunosorbent assay (ELISA) for GCSF at various time (0, 0.5, 1, 1. 5, 2, 3, 6, and 10 hrs)In the first day's experiment, that is, using a new infusion set (container, line and filters), the GCSF levels in the upper and lower part of filter and in the collecting vessels were very low during to 0.5 to 1.5 hrs after operating. However their levels roughly recovered until the container's during 3 to 6 hrs. A marked adsorption of G-CSF was also observed not only into the container and filters but also into the line. Notably the recovery of G-CSF was improved in the second day's experiment.It is necessary for physicians to give consideration to these adsorptions and to the loss of G-CSF when a small amount of G-CSF is dissolved for treatment by continuous intravenous infusion.

G‐CSF enhances the immunoglobulin generation rather than the proliferation of human B lymphocytes

The effect of granulocyte-colony stimulating factor (G-CSF) on human B-cell function was studied in in vitro cultures. G-CSF alone had no effect on the proliferative response of resting B cells, but it slightly enhanced the proliferative response of these cells in the presence of polyclonal B-cell mitogen, Staphylococcus aureus Cowan strain I (SAC) at concentrations of 0.2 to 25 micrograms/ml (1.5-fold increase in the DNA synthesis). In contrast, immunoglobulin (Ig) secretion of activated B cells was increased approximately three-fold to four-fold by adding G-CSF to the cultures. The neutralization of G-CSF bioactivity with anti-G-CSF antibody abrogated this effect. Though cytoplasmic Ig-positive cells or plasma cell marker-positive cells did not change, the expression of IgM mRNA in antibody-producing B cells increased in the presence of G-CSF in the cultures. Interestingly, human B lymphocytes are shown to express the binding to biotin-conjugated G-CSF preparation, but not to biotin-conjugated GM-CSF preparation when examined by flow cytometry. These data suggest that G-CSF may influence B-cell function in special circumstances.