G-CSF Group Research Articles

Granulocyte colony-stimulating factor increases the therapeutic efficacy of bone marrow mononuclear cell transplantation in cerebral ischemia in mice

BackgroundBone marrow mononuclear cell (BMMC) transplantation is a promising therapy for cerebral ischemia; however, little is known if its therapeutic efficacy may be improved by co-administration of potential modulatory factors in vivo. To explore this possibility, the present study examined the effect of BMMCs and G-CSF on cell proliferation, early neuronal development and neurological function recovery in experimental cerebral ischemia relative to controls that received neither treatment.ResultIschemia/infarct area was significantly reduced in BMMCs+G-CSF group relative to animal groups treated with BMMCs only, G-CSF only or saline. Transplanted BMMCs were found to colocalize with the proliferative cell nuclear antigen (PCNA) and the immature neuronal marker doublecortin (DCX). The BMMCs+G-CSF group showed increased numerical density of cells expressing PCNA and DCX, improved performance in adhesive sticker removal test and reduced neurological function severity scores relative to other groups in a time-dependent manner.ConclusionBMMCs and G-CSF co-administration exhibits synergistic beneficial effect over time. This effect could be at least partially related to increased proliferation and differentiation of bone marrow stem cells and enhanced host brain regeneration and functional recovery. The results suggest that G-CSF can increase the therapeutic efficacy of BMMCs transplantation in an experimental mouse model of cerebral ischemia.

Pretreatment of IL-2 influences the proliferation and polarization of mice splenic lymphocyte following mobilization with G-CSF

Objective To explore the influence of IL-2 pretreatment on splenic lymphocyte following mobilization with G-CSF, which may provide a new approach to attenuate acute graft-versus-host disease (GVHD). Methods Splenic cells and na(i)ve CD4+T cells from C57BL/6N mice receiving G-CSF-mobilized were pretreated with IL-2(50 U/ml) , and cocultured with the allogeneic antigens from BALB/c mice. The proliferation responses and the polarization of T cells were determined. C57BL/6N mice were randomly divided into 4 groups: G-CSF + IL-2 group, G-CSF group, IL-2 group , control group. Results Compared with the control group, IL-4 increased obviously while IFN-γ decreased significantly in the group of G-CSF + IL-2. The proliferation responses were also suppressed in vitro. Conclusion The proliferation responses of splenic cells and naive CD4 + T cells from C57BI/6N mice receiving G-CSF-mobilized to the allogeneic antigens were significantly abrogated by the pretreatment with IL-2, T cells were polarized toward the production of type-2 cytokines. The combination of G-CSF and IL-2 is potentially synergetic in the induction of T lymphocyte immune tolerance. Key words: Interleukin-2; Granulocyte colony-stimulating factor; Cell proliferation; Cell polarization; Mixed lymphocyte reaction

Etoposide plus G-CSF priming compared with G-CSF alone in patients with lymphoma improves mobilization without an increased risk of secondary myelodysplasia and leukemia

The use of etoposide (VP-16) for stem cell mobilization has been reported as a significant risk factor for the development of therapy-related myelodysplasia/therapy-related AML (tMDS/tAML) after transplantation. We compared the safety and effectiveness of VP-16+G-CSF (VP+G) to G-CSF alone for PBPC mobilization in patients with non-Hodgkin's lymphoma and Hodgkin's lymphoma who underwent autologous transplantation at the Cleveland Clinic and Ohio State University. In the VP+G group, median total CD34+ cells collected were 9.34 × 10(6) per kg (range 0.97-180.89), with 42% of all patients having adequate (2 × 10(6) cells per kg) CD 34+ collection after 2 days of apheresis compared with a median in the G-CSF group of 3.83 × 10(6) per kg (range, 0.72-50.38), with only 16% patients having adequate collection after 2 days (P<0.001). tMDS/tAML occurred in 15 patients (2.3%) in the VP+G and in 12 patients (3.8%) receiving G-CSF alone. (P=0.62). Increased number of days of apheresis was associated with the risk of tMDS/tAML (hazard ratio (HR) 1.19, 95% confidence interval (CI) 1.08-1.30, P<0.001). Priming regimen was not a significant variable for relapse-free survival or OS. The addition of etoposide significantly improves the effectiveness of mobilization at the cost of an increased incidence of neutropenic fever though with no mortalities. There is no evidence of increased incidence of tMDS/tAML in patients receiving VP+G compared with those mobilized with G-CSF alone.

Neutropenia and G-CSF Use In Patients with Relapsed/Refractory Multiple Myeloma Treated with Lenalidomide Plus Dexamethasone

AbstractAbstract 5034 IntroductionLenalidomide is approved in relapsed/refractory (rel/ref) MM based on 2 large phase III trials (Dimopoulos NEJM 2007; Weber NEJM 2007). In these trials, grade 3–4 neutropenia was common (29-41%) and was a main cause for lenalidomide (len) dose reductions despite mandated use of daily GCSF as first-step neutropenia management. Optimal management for len-induced neutropenia is unclear. At our institution, 216 patients (pts) with rel/ref MM were treated on the MM016, an Expanded Access Program (EAP), using len and dexamethasone (dex) at the same dose/schedule as that of the randomized trials. Due to limited access, GCSF was not mandated for grade 3–4 neutropenia, though we developed an intermittent schedule of GCSF (300ug SC 2–3 doses/week for weeks 3 and 4 of each 28 day cycle), typically continued into subsequent cycles to prevent recurrence. We aimed to avoid len dose reduction to maximize efficacy. In this retrospective analysis, our neutropenia management approach is evaluated, focusing upon recurrent neutropenia, infections, len dose-intensity, response, survival outcomes. MethodsFrom 2005–2008, 216 rel/ref MM pts were treated on the EAP at our center. Similar to the phase III studies, len was initiated at 25mg OD × 21days with dex 40mg days 1–4, 9–12, 17–20 every 28 day cycle. Recurrent neutropenia and infectious complications were reviewed. Comparison of baseline variables and neutropenia/survival was performed between pts receiving GCSF (group 1) and those not (group 2). OS and PFS were estimated by Kaplan-Meier curves and log rank test. ResultsNeutropenia and G-CSF use: Of 216 pts treated with len, 117 pts (54.2%) received GCSF for grade 3–4 neutropenia (group 1); 99 pts (45.8%) did not (group 2). For group 1, the first episode of grade 3–4 neutropenia occurred early (median cycle 2, range 1–20) with GCSF started at median cycle 3 (range 1–19). Although most pts continued GCSF prophylactically into subsequent cycles, almost half (58 pts; 49.6%) recurred with ≥1 episode of grade 3–4 neutropenia, 18 pts (15.4%) ≥3 episodes. Despite GCSF use, pts in group 1 had more grade 3–4 infections (47.0% vs. 25.5%, p=0.002) and hospitalization due to infection (40.2% vs. 25.3%, p=0.021). In addition, len dose reductions were frequent in group 1 (40.2% vs 16.2%; p<0.001). As a surrogate for dose-intensity, the proportion of cycles at full-dose without delay was lower in group 1 (0.67 vs. 0.96, p<0.001). However, group 1 pts received more lines of prior therapy than group 2 (median 3 vs. 2, p=0.021), had lower baseline platelets (<100/uL in 41.0% vs. 23.2%, p=0.006) and neutrophils (<2.0 bil/L in 41.0% vs. 20.2%, p<0.001). All other baseline characteristics were similar between groups. Responses and survival:Pts receiving GCSF remained on len for longer (median duration 10.3 vs 3.7 mos; p=0.01), with primary causes for len discontinuation in both groups due to disease progression (65.2%) and toxicity (15.2%). Increased responses (66.7% vs. 45.5%, p=0.002) and improved quality of response (CR/VGPR 25.6% vs. 14.1%, p=0.03) were seen in the GCSF group. G-CSF use was associated with significantly longer PFS (9.1 vs. 4.0 mos, p=0.048), though the OS between groups was not statistically different (20.9 vs. 13.7 mos; p=0.28). On multivariate analysis, G-CSF support was associated with decreased risk of death (HR 0.46, 95% CI 0.31–0.67, p<0.0001). Conclusions:1. Not unexpectedly, pts requiring GCSF were predisposed to len-induced neutropenia due to impaired marrow reserve from heavy pretreatment. Contrary to our predictions, our intermittent dosing schedule of GCSF did not significantly reduce rates of subsequent neutropenia, severe infections, or lenalidomide dose reduction. More intensive GCSF schedules than that used at our institution may be required.2. However, GCSF use is associated with longer duration on len therapy, likely leading to observed improvements in responses, quality of response, and PFS. This suggests that those pts with compromised marrow reserve benefit from dose reductions and striving for full dose-intensity is not necessary.3. Alternatively, given that GCSF is an independent predictor for prolonged PFS, one may hypothesize that GCSF has effects unrelated to its putative role in reducing neutropenia and may perhaps be related to its anti-inflammatory and immunomodulatory effects. Further studies evaluating the mechanisms and interactions of GCSF with len may provide insights. Disclosures:Reece:Celgene: Honoraria, Research Funding. Trudel:Celgene: Honoraria. Kukreti:Celgene: Honoraria. Chen:Celgene Corporation: Consultancy, Honoraria, Research Funding.

Efficacy of the coadministration of granulocyte colony-stimulating factor and stem cell factor in the activation of intrinsic cells after spinal cord injury in mice

Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic cytokine that induces undifferentiated stem cells from the bone marrow (BM) into the peripheral blood. Stem cell factor (SCF) is also a hematopoietic cytokine that stimulates the differentiation and proliferation of neural stem cells and has neuroprotective effects. In cerebrally infarcted mice, the combination of G-CSF and SCF promotes the differentiation of BM-derived cells into neural cells, stimulates the proliferation of intrinsic neural stem cells, and improves motor function. The object of this study was to investigate the effects of these cytokines on BM stem cells, intrinsic cells, and motor function recovery in spinal cord-injured mice. For marking BM-derived cells, the authors induced contusive spinal cord injury in mice transplanted with BM cells from green fluorescent protein (GFP)-transgenic mice after whole-body irradiation. These mice were treated with G-CSF and SCF in the subacute injury phase. Bromodeoxyuridine (BrdU) was injected into these mice to label proliferating cells. The cell numbers and phenotype of the BM-derived cells were evaluated, and the change in intrinsic cells (proliferation, accumulation, and differentiation) was noted using immunohistological analysis at 4 weeks postinjury (wpi). A behavior analysis was conducted until 12 wpi using the Basso, Beattie, Bresnahan locomotor rating scale. In the SCF + G-CSF group, improvement in hindlimb motor function was significantly greater than in the SCF group, G-CSF group, and sham-treatment (vehicle) group after 8 wpi. At 4 wpi, the number of GFP+ BM-derived cells induced in the lesion did not significantly differ between groups. At 4 wpi, the authors evaluated perilesional GFP− intrinsic spinal cord cells. The number of GFP− and F4/80+ cells was significantly greater in the SCF + G-CSF group than in the other 3 groups. As compared with the sham group, the number of NG2+/BrdU+ cells was significantly increased in the SCF + G-CSF group. In this study, the combined administration of SCF and G-CSF in traumatic spinal cord injury not only improved motor function, but also induced the accumulation of intrinsic microglia and the active proliferation of intrinsic oligodendrocyte precursor cells.

Use of G-CSF in Patients with Severe Aplastic Anemia Treated with ATG and Cyclosporine Increases Neutrophils and Decreases Infection Rates and Hospitalization Days but Does Not Improve Long-Term Outcome: Results of a Prospective, Randomized Clinical Trial of the EBMT.

Abstract 496▪▪This icon denotes an abstract that is clinically relevant.Immunosuppression (IS) with ATG and Cyclosporine (CSA) is the treatment of choice for patients with severe aplastic anemia (AA) not eligible for hematopoietic stem cell transplantation. The role of additional growth factors such as G-CSF on long-term outcome is still a matter of debate. The Aplastic Anemia Working Party of the EBMT conducted a randomized controlled study comparing IS with ATG and CSA with or without G-CSF and with or without early retreatment at day 120 in case of nonresponse. The study was designed to identify a difference of 10% for overall survival (OS) and event free survival (EFS). Three hundred forty subjects were to be enrolled. From 2002 to 2008, 205 patients were randomized, but 13 had to be excluded (1 incorrect diagnosis; 12 no follow-up data); 192 were evaluable for analysis (95 with G-CSF; 97 without G-CSF). The median age of the patients was 46 years (2-81), 95 (49%) were males, 69 (36%) had very severe AA. There was no difference between both groups in respect of age, sex, severity of the AA, and number of transfusions before treatment. Horse ATG (Lymphoglobulin) was administered (15mg/kg/BW/d ×5 days), and CSA given orally at a dose of 5mg/kg/d. Patients randomized to receive G-CSF were given glycosylated rHuG-CSF from day 8 to 120 (150mg/m2/d, sc).OS at 6 years was 75%, it was 82% for patients with severe AA and 66% for patients with very severe AA (P=0.001). Survival decreased with increasing age from 100% (age <20years) and 92% (20-40 years) to 71% (40-60 years) and 56% (>60 years) (P<0.001). There was no difference in OS (P=0.64) and EFS, defined by death, need for transplantation, relapse and non response as events (P=0.358) between the study arms at 6 years. This was the case for the entire cohort as well as when stratified according to age and AA severity (Table).Results of 192 patients treated with or without G-CSFall patients n=192no G-CSF n=95G-CSF n=97P-valueOS at 6 yearsall patients (%)75 ±477 ±576 ±50.64Severity (%)- SAA82 ±485 ±580 ±60.61- vSAA66 ±665 ±868 ±90.84Age groups (%)- < 20 years100100100-- 20 – 40 years92 ±489 ±696 ±40.39- 40 – 60 years71 ±871 ±972 ±130.34- > 60 years56 ±858 ±1157 ±100.78EFS at 6 yearsall patients (%)41 ±438 ±646 ±60.36Severity (%)- SAA44 ±539 ±849 ±70.64- vSAA39 ±638 ±842 ±90.45Age groups (%)- < 20 years58 ±952 ±1367 ± 120.65- 20 – 40 years49 ±946 ±1257 ± 120.98- 40 – 60 years39 ±947 ±1030 ± 150.79- > 60 years29 ±617 ±840 ± 110.10Deaths442321Cause of death- infection241212- bleeding743- MDS/AML2110.31- solid tumor201- cardiovascular422- TRM440- unknown111late deaths (>3y)5/390/205/190.01Infections and hospitalization during first 90 daysPatients with infection per period of 30 days- all patients30%36%24%0.006- severe AA27%24%30%0.30- very severe AA44%52%33%0.01Hospitalization days4261/113102192/56702067/56400.03The median number of neutrophils was significantly higher between day 30 and 240, in the treatment group but this difference did not persist to day 360, when G-CSF was stopped. There were fewer infections (36% no G-CSF; 24% with G-CSF; P=0.006), and less days of hospitalization during the first 90 days (P=0.03) in the group of patients with G-CSF, mainly for patients with very severe AA. During the study period 44 patients died. The most common cause of death was infection (55%).There was no difference in deaths and causes of death between both treatment groups. However, there were more late deaths (>3 years) in the G-CSF group (P=0.01). There was no difference in respect of response rates between both groups. Overall 73% of patients with and 62% without G-CSF did respond to IS (P=0.488). Response rates at days 30, 60, 90 120, 150, 180, 240 and 360 were similar between the treatment groups. Fifty seven patients did not respond to first-line therapy, and 31 patients relapsed during the first year of treatment, without any difference between both groups.In conclusion, G-CSF given together with standard IS increases neutrophil counts, and decreases rate of infections and days of hospitalization, mainly in patients with very severe AA. G-CSF does not improve long-term outcome and has no impact on OS, EFS, remission, death and relapse rates. G-CSF may decrease infection risks when used as an adjunct to IS therapy. This has to be weighed against possibly higher risks of MDS/AML, as suggested by previous studies. Disclosures:Schrezenmeier:Alexion: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.

G-CSF Mobilized Versus Conventional Donor Lymphocytes in the Treatment of Hematological Malignancies.

Abstract 4301 IntroductionDonor lymphocyte infusions (DLI) can reverse relapse of hematologic malignancy following allogeneic hematopoietic stem cell transplant (HSCT) in some cases. G-CSF is used to mobilize progenitor cells for transplantation and at our center excess cells have been cryopreserved and subsequently used as DLIs. Little is known regarding the effectiveness of G-CSF mobilized cells as DLIs in the treatment of hematological malignancies. We conducted a retrospective study comparing the overall survival in patients treated with G-CSF mobilized vs. conventional DLI for hematological malignancies. Patients and MethodsAfter IRB approval, we conducted a retrospective chart review of 67 patients who received DLI treatment of management of hematological malignancies. ResultsMean age was 48.5 (range 2-78). 41 (62%) patients had AML and 26 (38%) had other hematological malignancies. Patients received 1-4 DLIs each; median was 1 DLI per patient. Median no. of CD3+ cells infused was 107/kg body weight (range 1- 265). 17 (25%) patients received a total of 32 G-CSF mobilized DLIs. 50 (75%) patients received 77 conventional DLI. 20 (29%) patients were treated with immunosuppressive agents at the time of DLI and 49 (73%) received chemotherapy in the 90 days prior to DLI. DLIs were administered for relapse or residual disease post-transplant in 62 patients. 5 patients received 7 DLIs because of poor engraftment. In total 38(56%) patients have died. Death was malignant disease related in 31/38(82%) patients, and caused by graft versus host disease (GVHD) in 5/38 (13%). GVHD deaths were 2/17 in the G-CSF and 3/50 in the conventional group (NS). Median survival after DLI was 213 days for the 67 patients and was similar for GCSF and conventional groups. 19/67 patients survived more than 1 year from the initial DLI. Death and GVHD were not predicted by diagnosis, donor type (related of unrelated), use of immunosuppressives at the time and DLI or number of infused donor CD3 cells. Time to first evidence of GVHD was similar between the two groups (p=0.35). Kaplan-Meier estimates of survival suggested that G-CSF mobilized DLI, while not statistically different from conventional DLI, showed a trend towards a survival advantage (p=0.08). This analysis is shown in the figure below. ConclusionIn this series of poor prognosis hematological malignancies, DLI was clearly lifesaving in a number of patients. G-CSF mobilized DLI appears to be at least as effective as conventional DLI for the treatment of patients relapsing after allogeneic blood stem cell transplantation. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Febrile Neutropenia, Mucositis and Duration of Hospitalization Are Reduced After Pegfilgrastim Following Autologous Peripheral Blood Stem Cell Transplantation (PBSCT) for Malignant Lymphopathies: Report of 683 PBSCTs From a Single Institution.

Abstract 3410Poster Board III-298Granulocyte colony-stimulating factor (G-CSF) has been shown to decrease time to neutrophil recovery following PBSCT. Now, 3 products are available: 2 standard G-CSFs (filgrastim and lenograstim), and pegfilgrastim (PegG-CSF). In order to determine whether a single subcutaneous injection of Peg-G-CSF is as effective as a daily injection of standard G-CSF, in terms of haematological recovery, febrile neutropenic episodes (FN), antibiotic usage, hospitalization duration, mucositis, progressive free survival (PFS) and overall survival (OS), we retrospectively analyzed series of 558 patients having myeloma and lymphoma who underwent 683 PBSCT between 2000 and 2008 in our institution. Statistical analysis included univariate, Wilcoxon and χ2 fisher tests, logrank test for PFS and OS, and Cox model for multivariate analysis. From January 2000 and April 2005, 359 patients received standard G-CSF (filgrastim or lenograstim) and from May 2005 to December 2008, 298 patients received PegG-CSF. 26 patients did not receive G-CSF for any reasons. 427 PBSCTs have been performed for multiple myeloma (MM) after high dose melphalan, 197 for Non Hodgkin Lymphoma (NHL) and 59 for Hodgkin lymphoma (HL) with BEAM conditioning regimen. 133 patients underwent 2 or 3 PBSCT (130 MM and 3 HL). The mean of CD34 dose infused was 5.2 106 /kg (1.2-26.9) with 96% of the grafts containing more than 2.5 106 CD34/Kg. The median number of days of standard G-CSF given to reach an absolute neutrophil count (ANC) ≥500/ml was 9 days (0-29). Median time to neutrophil engraftment (ANC of 500/ml) was 11 days (5-30) in each group. The platelet recovery (platelet>20 000/mL) was 10 days (0-54) in each group. The platelet and RBC transfusion requirement are stastitically lower in the PegG-CSF than in the standard G-CSF group. As listed on the table, we have analyzed the following parameters for all patients: number of FN and their beginning and duration, number of antibiotic lines, duration of hospitalization, duration of mucositis, and the percentage of grade III and IV mucositis.Median, (range)Peg G-CSFstandard G-CSFNone G-CSFN298359p26FN rate (%)91.796.4<0.0196.16Duration of FN (days)2 (0-27)2 (0-18)0.622 (0-13)First day of FN (day)5 (-3-16)4 (-5-17)<0.014 (0-11)Number of antibiotic lines1 (1-5)2 (0-5)<0.013 (0-4)Duration of hospitalization (days)17 (8-62)19 (8-65)<0.0124 (18-35)Duration of mucositis (days)0 (0-75)6 (0-60)<0.017.5 (0-10)Mucositis grade III, IV (%)17.740.28<0.01NDThe same significantly differences are observed in MM, NHL and HL patients. The use of standard G-CSF or PegG-CSF did not modify OS at 1 and 5 years for both NHL and HL patients. In MM population PFS was unmodified but OS appeared better in the PegG-CSF group compared to standard G-CSF: respectively 1 y OS at 1 year, 96% versus 92%, OS at 5 years: 79% versus 53% (p= 0.034). Such a difference could be explained by the early use of bortezomib regimen for induction therapy before PBSCT more frequently in PegG-CSF group and this feature has been analyzed. Among patients undergoing autologous stem cell transplantation the use of Peg G-CSF seems to show an advantage in terms of duration of hospitalization and reduce the percentage of grade III, IV mucositis and the number of febrile neutropenic episodes. Disclosures:No relevant conflicts of interest to declare.

IGF-1 and G-CSF complement each other in BMSC migration towards infarcted myocardium in a novel in vitro model

Stem cell capability enhanced with cytokine administration is a promising treatment for myocardial infarction. Bone marrow stem cells (BMSCs) were isolated from C57BL/6 mice (8-12 weeks old) expressing GFP and characterized with c-kit and CD34. Infarcted heart tissue fragments were placed into dishes with BMSCs and medium supplemented with G-CSF, SCF, IGF-1 or combinations thereof were given to the BMSC-infarcted myocardium in vitro model. The IGF-1-G-CSF group showed significantly higher migration (67.7% +/- 2.6) of c-kit(+) BMSCs towards the ischemic tissue and expressed MEF-2 (43.7% +/- 1.7). Of the single treatment groups, the G-CSF group demonstrated significantly higher migration of c-kit(+) BMSCs (60.5 +/- 2.7) with MEF-2 expression (38.7 +/- 1.4). IGF-1 complements G-CSF and was relatively more significant in its effects on BMSC migration and cardiac lineage commitment towards ischemic heart tissue.

Erythropoietin Enhances Arterioles More Significantly Than it Does Capillaries in an Infarcted Rat Heart Model

Erythropoietin (Epo) is a hormone which regulates erythrocyte production. It has recently become known that Epo enhances angiogenesis. However, since shear stress is an initiator of arteriogenesis, this increase with Epo may be due to increased shear stress from erythrocytosis. To clarify this, we compared the effects of Epo on both angiogenesis and arteriogenesis. Myocardial infarction was induced by LAD ligation in Wistar rats (Epo, G-CSF and control). Epo (1,000 IU/kg) was administered immediately after ligation of the LAD. G-CSF was administered at 100 microg/kg/day for 5 days after the coronary ligation. Four weeks later, coronary angiography was performed using synchrotron radiation coronary micro-angiography with a Langendorff apparatus. The number of vessels was investigated by microscopy. The numbers of capillaries and arterioles (> 100 microm in diameter) were measured. Microscopical examination: Capillary density in the twilight zone was 95 +/- 19 in the control group, 126 +/- 24 in the G-CSF group, and 142 +/- 32 in the EPO group (control versus Epo: P < 0.005, control versus G-CSF: P < 0.05). Arteriole numbers were 4.3 +/- 0.2 in the control group, 6.9 +/- 1.0 in the G-CSF, and 11.8 +/- 0.6 in the Epo group (control versus Epo: P < 0.00001, G-CSF versus Epo: P < 0.00001, control versus G-CSF: P < 0.00001). The ratios of arterioles and capillaries were 0.048 +/- 0.013 in the control group, 0.057 +/- 0.016 in the G-CSF group, and 0.088 +/- 0.019 in the Epo group (control versus Epo: P < 0.0005, G-CSF versus Epo: P < 0.05). Angiography: The number of crossing arterioles in the 2 mm lattice was 5.4 +/- 1.7 in the Epo group and 3.8 +/-0.4 in the control group (P < 0.05). The gray scale values for the evaluation of capillaries was 128 +/- 3.7 and 119 +/- 2.1 in the Epo and control groups, respectively (P < 0.00005). Epo enhanced arterioles more significantly than it did capillaries in this infarcted rat heart model.

Mobilization effects of G‐CSF, GM‐CSF, and darbepoetin‐α for allogeneic peripheral blood stem cell transplantation

The effects of GM-/G-CSF and darbepoetin-alpha on stem cell mobilization were investigated. From February 2005 to March 2007, 30 allogeneic sibling donors were randomly assigned to a G-CSF group (5 microg/kg/day for 5-7 days) or triple group (GM-CSF 10 microg/kg/day on 1st and 2nd day, G-CSF 5 microg/kg/day for 5-7 days, and darbepoetin-alpha 40 mg on 1st day). The MNCs and CD34(+) cells were not different between the two groups, although the doses (x10(8)/kg of recipient body weight) of CD3(+) cells (3.64 +/- 1.75 vs. 2.63 +/- 1.36, P = 0.089) and CD8(+) cells (1.07 +/- 0.53 vs. 0.60 +/- 0.30, P = 0.006) were lower in the triple group. The engraftments, frequency of RBC transfusions, and hemoglobin recovery were not different between the two groups. The cumulative incidence of overall and Grades II-IV aGVHD was 64.3% vs. 61.1% and 25.9% vs. 27.1% in the G-CSF and triple regimen group, respectively, whereas the cumulative incidence of cGVHD was 20.8 +/- 1.3% and 24.4 +/- 1.7%, respectively. In conclusion, the triple regimen did not seem to be superior to G-CSF alone in terms of the CD34+ cell dose, hemoglobin recovery, and GVHD. However, the CD8+ cell count was significantly lower in the triple regimen group. The role of a lower CD8+ cell count in the graft may need to be elucidated in the future.

Pegfilgrastim and daily granulocyte colony-stimulating factor: patterns of use and neutropenia-related outcomes in cancer patients in Spain - results of the LEARN Study

Daily granulocyte colony-stimulating factors [(G-CSFs); e.g. filgrastim, lenograstim] are frequently used to reduce the duration of chemotherapy-induced neutropenia (CIN) and the incidence of febrile neutropenia (FN) in cancer patients. A pegylated formulation of filgrastim, pegfilgrastim, which is administered once per cycle, was introduced in Spain in 2003. LEARN was a multi-centre, retrospective, observational study in Spain comparing patterns of use of daily G-CSF and pegfilgrastim, and CIN-related outcomes in adults with non-myeloid malignancies receiving myelosuppressive chemotherapy. Outcome measures were the percentage of patients receiving G-CSF for primary prophylaxis versus secondary prophylaxis/treatment, duration of treatment with G-CSF and incidence of CIN-related complications. Medical records from consecutive patients with documented pegfilgrastim (n = 75) or daily G-CSF (n = 111) use during 2003 were included. The proportion of patients receiving primary or secondary prophylaxis was comparable between the pegfilgrastim (39 and 48% respectively) and daily G-CSF (40 and 48% respectively) groups. However, there was a trend towards less frequent use to treat a neutropenic event such as FN or neutropenia in the pegfilgrastim group (17 versus 30% with daily G-CSF). Chemotherapy-induced neutropenia-related complications were less frequent in patients receiving pegfilgrastim (e.g. FN 11 versus 24% with daily G-CSF). This is the first study to show the potential benefits of pegfilgrastim over daily G-CSF in Spanish clinical practice.

GCSF Alone or in Combination with Cyclophosphamide (C) or GM-CSF for Hematopoietic Stem Cell Mobilization (HSCM) for Autologous Peripheral Blood Stem Cell Transplant (AHPCT): A Retrospective Review of a Single Center Experience

The mechanisms of HSCM are not completely understood and involve the action of multiple cytokines and adhesion molecules. Historically, HSCM for AHSCT has been done using high dose chemotherapy, usually C, with growth factor (G-CSF or GM-CSF). However, the need for the chemotherapy in this instance has not been defined. We, retrospectively compared three HSCM approaches; C and GSCF, G-CSF and GM-CSF, and G-CSF alone in our institution from 2005–2007. Results: C (2.5gm/m2) with G-CSF (10mcg/kg) was used in 25 patients (pts) (n=18 multiple myeloma [MM], n=5 non-Hodgkin's lymphoma [NHL], n=1 Hodgkin's disease [HD] and n=1 systemic sclerosis). G-CSF 5mcg/kg with GM-CSF 5mcg/kg was used for HSCM in 10 pts (n=8 NHL and n=2 MM). G-CSF alone (10mcg/kg for at least 4 days) was used in 49 pts (n=28 MM, n=13 NHL, n=2 HD and n=6 other diagnosis). The age range was 35–72 in the C and G-CSF group, 30–69 in the G-CSF and GM-CSF, and 25–72 in the GCSF alone group. The median CD34+cells/kg collected was 6.4×106 (range 0.97–131), 12×106 (range 0.23–6.7), and 4.59×106 (range 0.66–7.38) in the C and GCSF, G-CSF and GM-CSF, and the G-CSF groups respectively. The collection sessions ranged from a median of 2.0 days (1 day in 44% pts, 2 days in 28%, 3 days in 16% and 4 days in 12% of pts) in the C and G-CSF group; to 2.5 days (1 day in 20% of pts, 2 days in 30%, 3 days in 30% and 4 days in 20% of pts) in the G-CSF and GM-CSF group; and 2 days (1 day in 22% of pts, 2 days in 37%, 3 days in 30% and 4 days in 10% of pts) in the G-CSF alone group. 3 pts (12%) failed HSCM with C plus GCSF, 1 subsequently collected with G-CSF alone. 5 pts (50%) failed HSCM with G-CSF +GMCSF, 2 of these had previously failed G-CSF alone, and 2 were re-mobilized with AMD 3100 protocol and 1 with C and GCSF. 6 pts (12%) failed HSCM with G-CSF alone, 3 subsequently underwent HSCM with higher dose of G-CSF and 2 were enrolled on AMD-3100 study. 2 pts (8%) with C and G-CSF HSCM developed neutropenic fever and needed hospitalization. All pts who had adequate HSCM (at least 2×10e6/kg) and subsequently underwent AHPCT had white blood and platelet engraftment. There was no difference in the day to engraftment based on the HSCM strategy. These data show that HSCM with G-CSF alone results in similar stem cell yield and stem cell collection sessions to C and G-CSF. Although the latter strategy achieved a higher median number of HSC (4.9 vs. 6.4 xe6/kg), this was not statistically significant (p= 0.08) in this group of pts. A higher percentage of pts achieved target stem cell collection in one day with C plus G-CSF (44%), than G-CSF alone (22%). On the other hand, the combination of C plus G-CSF carries risks related to high dose chemotherapy and necessitates hospitalization for its administration and management of neutropenic fever. The G-CSF+GM-CSF combination was the least effective in adequate HSCM. Analysis of DFS in the three mobilization strategies is pending.

Effects of granulocyte-colony stimulating factor on the repair of balloon-injured arteries

The dysfunction of vascular endothelial cells plays a key role in starting arterial restenosis. The circulating endothelial progenitor cells (EPCs) can be mobilised by cytokines and are recruited to sites of injury, where they may participate in intima repair and the recovery of endothelial function. In the present study, we examined the hypothesis that mobilisation of EPCs by exogenous recombinant human granulocyte-colony stimulating factor (rhG-CSF) can promote vascular proliferation, reduce vascular inflammation and decrease the rate of restenosis. Male rats were randomly divided into the rhG-CSF and control groups. The animals were injected daily with 30 microg/kg rhG-CSF or 0.9% NaCl subcutaneously for 7 days, then the animals underwent balloon angioplasty of the common carotid artery. The animals were euthanased at 2 or 4 weeks after injury, and the carotid arteries were harvested and processed for immunohistochemistry, scanning electron microscopy (SEM), morphometric analysis of endothelialisation and neointimal formation at 1 hour, and 3, 7, 14 and 28 days after injury. Immunohistochemistry for proliferation cell nuclear antigen (PCNA) and reverse transcriptase polymerase chain reaction (RT-PCR) for endothelial nitric oxide synthase (eNOS) mRNA were also conducted for evaluating the proliferation of cells of the vessel wall and the possible mechanism of the repairing. Four weeks after balloon damage, SEM showed increased re-endothelialisation of the denuded vessels in the G-CSF treated animals compared with the control animals [(60.6 +/- 7.3)% versus (41.6 +/- 3.3)%, p < 0.01]. Re-endothelialisation was paralleled by a decrease in inflammation in the vessel wall. Histological examination showed that neointimal formation, vascular smooth muscle cells and fibrous tissue of the G-CSF group were less than those of the control group. Morphometry showed the lumen area of the G-CSF group was larger than that of the control group, and the neointimal area and percent of intimal hyperplasia were significantly smaller than those of the control group. Immunohistochemical staining for PCNA positive cells was less in the G-CSF treated animals compared with the control animals (42.9% versus 72.8%, p < 0.01). rhG-CSF-induced mobilisation of EPCs regenerated endothelium and inhibited neointimal hyperplasia after vascular injury. This cytokine therapy may be a feasible strategy for the promotion of re-endothelialisation after angioplasty.

A Randomized Controlled Trial of Granulocyte Colony-Stimulating Factor for the Treatment of Severe Sepsis Due to Melioidosis in Thailand

Melioidosis is a tropical infectious disease associated with significant mortality. Most deaths occur early and are caused by fulminant sepsis. In this randomized, placebo-controlled trial, we assessed the efficacy of lenograstim (granulocyte colony-stimulating factor [G-CSF], 263 mu g per day administered intravenously) in ceftazidime-treated patients with severe sepsis caused by suspected melioidosis in Thailand. Over a 27-month period, 60 patients were enrolled to receive either G-CSF (30 patients, 18 of whom had culture-confirmed melioidosis) or placebo (30 patients, 23 of whom had culture-confirmed melioidosis). Mortality rates were similar in both groups (G-CSF group, 70%; placebo group, 87%; risk ratio, 0.81; 95% confidence interval, 0.61-1.06; P=.2), including among patients with confirmed melioidosis (83% vs. 96%; P=.3). The duration of survival was longer for patients who received G-CSF than for patients who received placebo (33 h vs. 18.6 h; hazard ratio, 0.56; 95% confidence interval, 0.31-1.00; P=.05). Receipt of G-CSF is associated with a longer duration of survival but is not associated with a mortality benefit in patients with severe sepsis who are suspected of having melioidosis in Thailand. We hypothesize that G-CSF may "buy time" for severely septic patients, but survival is more likely to be improved by management of associated metabolic abnormalities and organ dysfunction associated with severe sepsis.

C‐terminal pentapeptide of osteogenic growth peptide regulates hematopoiesis in early stage

Osteogenic growth peptide (OGP) was characterized in regenerating bone marrow, which can increase osteogenesis and hematopoiesis. The carboxy-terminal pentapeptide is a naturally occurring human and murine mitogen equipotent to OGP. In this study, we evaluated the potential role of OGP10-14 in regulation of hematopoiesis in human hematopoietic stem cells and animal model. Our results showed CD34+ stem cells from umbilical cord blood (UBC) were significantly increased in OGP10-14 treated samples, which is nearly equivalent to the results obtained from the combinations of IL3, IL11, G-CSF, and EPO group. OGP10-14 can also stimulate the differentiation of stem cells from bone marrow at the level of noncommitted progenitor stem cells, thus increasing the number of reconstituted red and white cells as well as platelets after injected i.m. everyday continuing for 5 days in hematopoietic function damage mice comparing with the OGP-untreated group. These data implicate that the role of OGP10-14 regulating hematopoiesis is in the early stage of the whole hematopoietic growth factors (HGFs) regulating network, just like the position of interleukin 13 in the hematopoiesis network.

Low doses of GM-CSF (molgramostim) and G-CSF (filgrastim) after cyclophosphamide (4 g/m2) enhance the peripheral blood progenitor cell harvest: results of two randomized studies including 120 patients

The use of a combination of G-CSF and GM-CSF versus G-CSF alone, after cyclophosphamide (4 g/m2) was compared in two randomized phase III studies, including 120 patients. In study A, 60 patients received 5 x 2 microg/kg/day of G-CSF and GM-CSF compared to 5 mug/kg/day of G-CSF. In study B, 60 patients received 2.5 x 2 microg/kg/day G-CSF and GM-CSF compared to G-CSF alone (5 microg/kg/day). With the aim to collect at least 5 x 10(6)/kg CD34 cells in a maximum of three large volume leukapherises (LK), 123 LK were performed in study A, showing a significantly higher number of patients reaching 10 x 10(6)/kg CD34 cells (21/29 in G+GM-CSF arm vs 11/27 in G-CSF arm, P=0.00006). In study B, 109 LK were performed, with similar results (10/27 vs 15/26, P=0.003). In both the study, the total harvest of CD34 cells/kg was twofold higher in G-CSF plus GM-CSF group (18.3 x 10(6) in study A and 15.85 x 10(6) in study B) than in G-CSF group (9 x 10(6) in study A and 8.1 x 10(6) in study B), a significant difference only seen in multiple myeloma, with no significant difference in terms of mobilized myeloma cells between G-CSF and GM-CSF groups.

ELYPSE 2: A prospective randomized trial comparing filgrastim (G-CSF) in primary and secondary prophylaxis in patients (pts) at high risk for febrile neutropenia (FN)

8614 Background: FN is a frequent life-threatening consequence of cytotoxic chemotherapy (CT). G-CSF reduces the risk of FN, but primary (1ry) prophylaxis using G-CSF may be cost efficient only if FN incidence is ≥20%. The identification of pts at high risk for FN with simple criterias would be useful in clinical practice. Here we report a randomized phase II trial comparing G-CSF in 1ry vs 2ry prophylaxis in a high risk group of pts (based on our risk model JCO 1996;14:737, Br J Cancer 2003;88:181). Methods: Pts ≥18 years with solid tumors or NHL at high risk for FN after CT were incluable: these were pts with day 1 or day 5 lymphocytes (Ly) count ≤700/μL AND “high risk CT” (HRCT) In pts with d1 and d5 Ly ≤700/μL, the observed incidence of FN was 40% and 66% respectively G-CSF (300 to 480μg/d from d6–12) was randomized either as in 1ry prophylaxis (Arm 1), or as 2ry prophylaxis after FN (Arm 2). Primary endpoint was the rate of grade 4 FN. Hypothesis was a reduction of 40% of the risk of FN with 1ry prophylaxis. Results: Between 03/97 and 12/04, 137 pts were included in 7 centers. The median age was 53 years (range 18–80) with 54% males. Most frequent tumors were sarcomas (36%), breast carcinomas (18%), lymphomas (15%), head and neck carcinomas (10%), and lung carcinomas (6%). 23% patients had PS&gt;1 at the first line of chemotherapy. No difference was observed in terms of duration of hospitalization or antibiotherapy. Median number of days of G-CSF administration was 14 days (0–24) vs 0 (0–17) days (p&lt;0.0000). After the 1st course, grade 4 FN was 38% in Arm 2 (2ry prophylaxis) and 25% in Arm 1 (1ry prophylaxis), showing a 34% reduction of FN in arm 1 (p=0.14): 1ry prophylaxis was associated with a significant reduction of FN using logistic regression (p=0.04). Incidence of FN after the 2 course in pts receiving 2ry prophylaxis was 22%. Among the subgroup of pts with PS&gt;2 and Ly ≤700/μL, (a group with a reported 20% risk for early death, Br J Cancer 2001;85:816), 2 of 8 patients (25%) died after the 1st course, vs 0/13 in the G-CSF group (p=0.05). This difference was not significant in the whole group. Conclusions: This study confirms that lymphopenic pts receiving HRCT are a high risk group of pts for FN for whom 1ry prophylaxis with G-CSF reduces the incidence of FN. [Table: see text]

Successful Engraftment of Autologous CD34+ Stem Cells after High-Dose Therapy and Fixed Dose (6 mg) Pegfilgrastim.

High-dose (HD) therapy plus Autologous Stem Cell Transplantation (ASCT) is a milestone treatment program for most hematologic malignancies. Daily subcutaneous injections of G-CSF (filgrastim/lenograstim) until ANC > 500/μl are routinely administered following ASCT, in order to accelerate hematopoietic recovery and reduce neutropenic complications. Pegfilgrastim has been shown to have similar efficacy when compared to G-CSF for chemotherapy-induced neutropenia, but little is known about its use in the ASCT setting. We used a 6 mg fixed dose of Pegfilgrastim on day +4 following ASCT in 46 patients (22 M/24 F; median age 56 yrs; r 22-70 yrs) with multiple myeloma (25 pts) and relapsed or refractory Hodgkin's and non-Hodgkin's lymphoma (21 pts). Patients received peripheral CD34+ stem cells (median number 4.4 x 106/Kg; r 1.8 – 11.8) harvested after mobilizing chemotherapy (cytoxan, vinorelbine/cytoxan, R-IEV, IGEV, R-ICE) and G-CSF. Standard conditioning regimens (HD-Melphalan or BEAM) were used. Engraftment results were compared to those from a historical control group of 182 patients (median age 56 yrs; r 16-74 yrs) who had received HD-Melphalan or BEAM and ASCT (median CD34+ cells 7.6 x 106/Kg, r 1.8–4.6) supported by G-CSF (5 μg/kg/day from day +5 until ANC>500/μl). Median number of days to ANC>500/μl were comparable between the Pegfilgrastim (10, r 8-15) and G-CSF (11, r 7–22) groups, as well as the median number of days to PLT>20,000/μl (Pegfilgrastim = 12, r 9–20 vs G-CSF= 12, r 7–29). Overall infectious rates, including FUO and documented infections, were of 48% and 39% for Pegfilgrastim and G-CSF groups, respectively (p=NS). Median number of days on i.v. antibiotics were 0 (r 0–18) and 6 (r 0–13) for the Pegfilgrastim and G-CSF groups, respectively. No significant differences in the incidence of bone pain, intensity of transfusion support and length of hospital stay were documented between the two groups. These data indicate that a fixed (6 mg) single-dose of Pegfilgrastim is safe and effective to accelerate engraftment after ASCT. No significant differences with G-CSF were apparent as to engraftment times and overall infectious complications. Growth factor costs were however in favor of Pegfilgrastim (Euro 690 vs 892 for a median of 10.5 G-CSF doses).

G-CSF Administration Increases Pulmonary Toxicity for Hodgkin's Disease Patients Treated with Bleomycin-Containing Chemotherapy.

Background: In Hodgkin's Disease (HD), G-CSF has been used to maintain maximum dose intensity by avoiding neutropenia and treatment delays. Pre-clinical data and anecdotal reports have shown an association between G-CSF administration and bleomycin pulmonary toxicity (BPT). This complication may cause access morbidity and have a detrimental impact on survival in this otherwise good prognosis population. We embarked on a retrospective analysis to determine if G-CSF increased the incidence of this complication or had an impact on outcomes.Methods: We reviewed all patients managed for HD at Mayo Clinic, Rochester, from January 1986 to February 2003. BPT was defined by the presence of pulmonary symptoms, bilateral interstitial infiltrates, and no evidence for an infectious etiology. Patients who received G-CSF support with one or more of their bleomycin-containing chemotherapy cycles were considered as being treated with G-CSF.Results: A retrospective review identified 141 patients with HD treated with a bleomycin-containing regimen. The mean age of patients at diagnosis was 34 years. Fifty four percent were males. The histology was nodular sclerosing in 85%. The Hasenclever index was 0 in 15%, 1 in 35%, 2 in 24%, 3 in 16%, 4 in 9%, and ≥ 5 in 1%. The G-CSF and non G-CSF groups were balanced for poor prognosis risk factors including Hasenclever index. Frontline chemotherapy included ABVD in 57%, MOPP-ABV(D) in 33%, COPP-ABV in 8%, BEACOPP in 3%, and Stanford V in 2%. Forty one percent of patients received radiation. The 5-year overall survival (OS) and progression free survival (PFS) for all patients were 87% and 79% respectively. G-CSF was administered to 52% (74/141) of patients. BPT occurred in 18% of patients (25/141). A higher rate of BPT was observed in patients receiving G-CSF, 26% (19/74) versus 9% (6/67) in non G-CSF patients (p=0.0141). In this population, G-CSF administration was the only factor associated with an increased risk of BPT. There was no difference between the two groups for BPT associated risk factors including radiation, bleomycin dose, renal insufficiency, smoking history, and underlying lung disease. Additionally these risk factors showed no correlation with the development of BPT. The mortality from BPT was 4.2% (6/141) in all patients and 24% (6/25) in patients who developed the pulmonary syndrome. Of the 6 patients who died acutely, 83% (5/6) received G-CSF. CR rates were equal between the G-CSF and non G-CSF groups at 92% and 94% respectively. Five year OS and PFS were equal between the G-CSF and non G-CSF groups at 82% versus 89% (p=0.473) and 74% versus 84% (p=0.152) respectively.Conclusion: G-CSF administration increases the incidence of pulmonary toxicity in HD patients treated with bleomycin based chemotherapy. The major impact was on treatment related morbidity as 5-year OS and PFS were equal between the G-CSF and non G-CSF groups.