Doses Of rhG-CSF Research Articles

Site-selective fatty acid chain conjugation of the N-terminus of the recombinant human granulocyte colony-stimulating factor.

The clinical application of the recombinant human granulocyte colony-stimulating factor (rhG-CSF) is restricted by its short serum half-life. Herein, site-selective modification of the N-terminus of rhG-CSF with PAL-PEG3-Ph-CHO was used to develop a long-acting rhG-CSF. The optimized conditions for rhG-CSF modification with PAL-PEG3-Ph-CHO were: reaction solvent system of 3% (w/v) Tween 20 and 30mM NaCNBH3 in acetate buffer (20mmol/L, pH 5.0), molar ratio of PAL-PEG3-Ph-CHO to rhG-CSF of 6:1, temperature of 20°C, and reaction time of 12h, consequently, achieving a PAL-PEG3-Ph-rhG-CSF product yield of 70.8%. The reaction mixture was purified via preparative liquid chromatography, yielding the single-modified product PAL-PEG3-Ph-rhG-CSF with a HPLC purity exceeding 95%. The molecular weight of PAL-PEG3-Ph-rhG-CSF was 19297Da by MALDI-TOF-MS, which was consistent with the theoretical value. The circular dichroism analysis revealed no significant change in its secondary structure compared to unmodified rhG-CSF. The PAL-PEG3-Ph-rhG-CSF retained 82.0% of the in vitro biological activity of unmodified rhG-CSF. The pharmacokinetic analyses showed that the serum half-life of PAL-PEG3-Ph-rhG-CSF was 7.404 ± 0.777h in mice, 4.08 times longer than unmodified rhG-CSF. Additionally, a single subcutaneous dose of PAL-PEG3-Ph-rhG-CSF presented comparable in vivo efficacy to multiple doses of rhG-CSF. This study demonstrated an efficacious strategy for developing long-acting rhG-CSF drug candidates.

Analysis of Agranulocytosis Time and Its Influencing Factors in Patients with Hematological Malignancies Treated with rhIL-11 combined rhG-CSF

To explore the intervention effect of recombinant human interleukin-11 (rhIL-11) and recombinant human granulocyte-colony stimulating factor (rhG-CSF) on the duration and severity of agranulocytosis in patients with hematological malignancies after chemotherapy, and to analyze the influencing factors. The data of hematological malignancy patients treated with rhIL-11 and rhG-CSF after chemotherapy in the hematology department of The First Hospital of Lanzhou University from July 2017 to July 2020 were collected retrospectively. The duration and differences of agranulocytosis in differeent groups were compared by univariate analysis, and the influencing factors of agranulocytosis duration were further analyzed by multiple regression analysis. The duration of agranulocytosis in 97 patients was 6.47±2.93 days. The results of univariate analysis showed that there were no statistical differences in the duration of agranulocytosis among patients with different sex, age, height, weight, body surface area, body mass index (BMI), dose of rhG-CSF, dose of rhIL-11, spontaneous bleeding after administration of rhG-CSF and rhIL-11, and the duration of agranulocytosis in patients with different red blood cell count (RBC), hemoglobin(HGB) level, platelet count (PLT) and absolute neutrophil count (ANC), before administration of rhG-CSF and rhIL-11. There were significant differences in agranulocytosis time among patients with different disease types, chemotherapy cycle, fever after rhG-CSF and rhIL-11 administration, and different white blood cell count (WBC) baseline level before rhG-CSF and rhIL-11 administration (P＜0.05). Compared with patients with acute lymphoblastic leukemia (ALL) and non-Hodgkin lymphoma (NHL), patients with acute myeloid leukemia (AML) had the longest duration of agranulocytosis, which was 7.07±3.05 d. Compared with patients with chemotherapy cycles of 4－6 and ≥7, patients with total chemotherapy cycle of 1－3 had the shortest duration of agranulocytosis, which was 5.25±2.48 d. Compared with patients without fever, patients with fever within 1 day after administration of cytokines and patients with fever within 2-5 days after administration of cytokines, the duration of agranulocytosis was the longest in patients with fever 6 days after administration of cytokines, which was 8.85±2.85 d. Compared with patients with WBC baseline <1.0×109/L, (1.0－1.9)×109/L and (2.0－3.9)×109/L, patients with WBC baseline ≥4.0×109/L had the shortest duration of agranulocytosis, which was 4.50±2.56 d. Multiple linear regression analysis showed that chemotherapy cycle, different fever after administration of rhG-CSF and rhIL-11, diagnosis of ALL and NHL, and WBC baseline level before administration of rhG-CSF and rhIL-11 were the influencing factors of the duration of agranulocytosis (P＜0.001). The risk of prolonged agranulocytosis is higher in patients diagnosed with AML, with more chemotherapy cycles, lower WBC baseline before cytokines administration and fever later after cytokines administration, which should be paid more attention to.

Analysis of Factors Influencing Autologous Peripheral Blood Stem Cells Mobilization in Patients with Lymphoma and Multiple Myeloma

To analyze the factors influencing the mobilization of autologous peripheral blood stem cells (auto-PBSCs) in patients with lymphoma and multiple myeloma, and provide reference for optimizing the autologous stem cell mobilization regimen. Clinical data of 33 multiple myeloma and lymphoma patients received auto-PBSCs mobilization in our center from January 2015 to December 2018 were collected, the correlation of mobilization failure rate with gender, age, courses of chemotherapy before mobilization, does of recombinant human granulocyte colony stimulating factor (rhG-CSF), type of disease, and chemotherapy regimen were retrospectively analyzed. Type of disease and course of pre-mobilization chemotherapy could affect the mobilization failure rate (P<0.05). The mobilization failure rate of lymphoma patients was 42.1%, which was significantly higher than 7.1% of multiple myeloma patients (P=0.026). The mobilization failure rate was higher in the group with chemotherapy courses≥5 before mobilization (P=0.016). Age, gender, dose of rhG-CSF, and chemotherapy regimen had no significant correlation with mobilization failure rate (P>0.05). Multi-course chemotherapy before collection and lymphoma patients are poor factors negatively impacting on auto-PBSCs mobilization.

Allogeneic Blood Stem Cell Transplantation: Considerations for Donors

AbstractAllogeneic transplantation of cytokine-mobilized peripheral blood stem cells (PBSCs) is now being increasingly performed, but safety considerations for hematologically normal PBSC donors have not been fully addressed. Progenitors are generally mobilized for collection from normal donors using recombinant human granulocyte colony-stimulating factor (rhG-CSF). Although the short-term safety profile of rhG-CSF seems acceptable, experience remains limited and its optimal dose and schedule have not been defined. Minimal data exist regarding long-term safety of rhG-CSF, primarily derived from experience in patients with chronic neutropenia or cancer. An “ad hoc” workshop was recently convened among a group of investigators actively involved in the field of allogeneic stem cell transplantation to discuss the safety issues pertaining to normal PBSC donors. There was agreement on the following points: (1) On the basis of available data, it appears that rhG-CSF treatment and PBSC collection have an acceptable short-term safety profile in normal donors. However, the need for continued safety monitoring was recognized. (2) rhG-CSF doses up to 10 μg/kg/d show a consistent dose-response relationship with the mobilization (and collection) of CD34+ progenitor cells, and this dose is acceptable for routine clinical use. Whether higher doses are superior (or cost effective) remains to be determined, and they may produce more severe side effects. The potential risks of marked leukocytosis (arbitrarily defined as a leukocyte count of more than 70 × 109/L) have been a concern, and rhG-CSF dose reduction is performed by many centers to maintain leukocyte counts below this level. (3) Transient post donation cytopenias, involving granulocytes, lymphocytes, and platelets, may occur and are at least partly related to the leukapheresis procedure. These are generally asymptomatic and self-limited; follow-up blood counts are not necessarily required. Reinfusion of autologous platelet-rich plasma should be considered for donors with expected postdonation thrombocytopenia (platelet count < 80 to 100 × 109/L). (4) Donors should meet the eligibility criteria which apply to donors of apheresis platelets, with the exception that pediatric donors may also be considered. Any deviation from these criteria should have supporting documentation. There is insufficient information at this time to clearly establish definite contraindications for PBSC collection in a hematologically normal donor. Potential contraindications include the presence of inflammatory, autoimmune, or rheumatologic disorders, as well as atherosclerotic or cerebrovascular disease. (5) The creation of an International PBSC Donor Registry is desirable to facilitate monitoring the long-term effects of the procedure. Individual institutions or donor centers are encouraged to establish their own PBSC donor follow-up system, preferably with a standardized approach to data collection.

The prophylactic and therapeutic effects of moxibustion combined with traditional Chinese medicine decoction for treating chemotherapy-induced myelosuppression in early-stage breast cancer: study protocol for a randomized controlled trial

BackgroundTraditional Chinese medicine (TCM) has a long history of use in breast cancer, but lacking systematic evidence to support its clinical benefits. In this study, we evaluated the prophylactic and therapeutic effects of moxibustion combined with decoctions for treating chemotherapy-induced myelosuppression (CIM) in early-stage breast cancer patients.MethodsThis is a randomized controlled clinical trial single-blinded for TCM decoction but not moxibustion. Patients are equally divided into the control group without decoction and moxibustion treatment (control), the decoction+moxibustion group (MD), and the placebo+moxibustion group (MP), according to the following stratification factors: age (below 40s, 40s, 50s, and 60s or above), chemotherapy regimen (anthracyclines, taxanes, anthracyclines+taxane, and others), and chemotherapy strategy (adjuvant and neoadjuvant). The TCM decoction is Wenshen Shengbai Decoction. The anticipated sample size is 462 cases (154 cases in each group). All participants are expected to treat with chemotherapy and recombinant human granulocyte colony-stimulating factor (rhG-CSF). The primary outcomes include the proportion of patients with relief of leukopenia and/or neutropenia, the myelosuppression-associated serious adverse event including grade 3–4 leukopenia and/or neutropenia, and febrile neutropenia, and the dose of rhG-CSF. The secondary outcomes include chemotherapy adherence, stratified analysis, adverse reactions, quality of life by EORTC Breast-Cancer-Specific Quality of Life Questionnaire including EORTC QLQ-C30 (V3.0) and QLQ-BR23, TCM Constitution, and 3-year disease-free survival and overall survival. Baseline information including age, surgical approach, chemotherapy regimen and strategy, pathological stage, and molecular subtype will be recorded.DiscussionThis will be the first randomized controlled trial to evaluate the efficacy of moxibustion combined with TCM decoction in treating CIM in early-stage breast cancer patients, aiming to standardize the TCM decoction and moxibustion method, thus providing evidence for its clinical benefit.Trial registrationchictr.org.cn ChiCTR-INR-16009557. Registered on 23 October 2016.

Bone pain associated with rhG-CSF or PEG-rhG-CSF in oncology patients.

e19186 Background: Recombinant human granulocyte colonystimulating factor (rhG-CSF) and glycoPEGylated G-CSF (PEG-rhG-CSF) are two kinds of drugs in reducing the incidence and duration of neutropenia in oncology patients treated with chemotherapy. Although the acting time is different, bone pain is one of the most common adverse events in rhG-CSF and PEG-rhG-CSF. The purpose of this study is to describe the occurrence and management of bone pain in oncology patients receiving rhG-CSF and PEG-rhG-CSF. Methods: Two hundred and forty patients with malignant tumor received rhG-CSF or PEG-rhG-CSF after chemotherapy were enrolled to finish questionnaires about the side effect of bone pain. The incidence, location, duration, degree and treatment of bone pain after rhG-CSF or PEG-rhG-CSF used were collected and analyzed. Results: A total of 240 patients enrolled and 56.3% (135) patients had bone pain after rhG-CSF or PEG-rhG-CSF delivered. The most common parts of bone pain were thigh (35.6%), back (29.6%), waist (25.9%), shoulder (23.7%) and chest (21.5%). The bone pain of 39.2% patients last for 72h and 27.2% patients last less than 72h. The duration time of bone pain was longer in PEG-rhG-CSF than rhG-CSF. There was no significant difference in the rate of pain occurrence and the proportion of severe pain between rhG-CSF and PEG-rhG-CSF. Only 22.2% patients took drugs to relieve bone pain. Downregulated the doses of rhG-CSF or PEG-rhG-CSF could significantly relieve bone pain. Besides bone pain, 34.0% patients suffered muscle ache which was the second most side effect and occurred more often than runny nose and fever. Conclusions: The percentage of bone pain occurrence are similar in rhG-CSF and PEG-rhG-CSF, but the duration time was longer in PEG-rhG-CSF. For most patients, bone pain need not to deal with, Dose reductions can effectively relieve the mild to severe pain.

Bioassay of Recombinant Human Granulocyte Colony Stimulating Factor (rhG-CSF) for Neutropenia Treatment in Male Sprague Dawley Rats

Background: Recombinant human granulocyte colony stimulating factor (rhG-CSF) is a first line therapy for neutropenia. However, it is less affordable for most patients in developing and poor countries. Therefore, biosimilar products are developed to suppress the cost of treatment, namely with rhG-CSF. This study aimed to explore the establishment of an affordable rhG-CSF that has similar potential to induce neutrophils recovery as the positive control.Materials and Methods: The rhG-CSF was expressed as inclusion body in Escherichia coli NiCo21(DE3). The inclusion body was then solubilized, refolded, purified and characterized prior to be used in the bioactivity assay. Cyclophosphamide-induced male Sprague Dawley rats were used as animal model and administered with rhG-CSF. Blood sample was collected at several points of time, before and after rhG-CSF treatments. Complete blood count and peripheral blood smear were conducted to investigate the activity of the rhG-CSF on each blood cells type, particularly neutrophil.Results: Specific activity on neutrophil proliferation was shown after treatments with our rhG-CSF and positive control. Positive control dose 40 mg/kg BW was statistically similar with that of the rhG-CSF dose 80 and 120 mg/kg BW. However, in neutropenic condition, recovery of neutrophil counts could not be achieved within 4 days of treatments. Thus, a longer treatment is needed to observe the activity of the rhG-CSF as an antineutropenia agent.Conclusion: The rhG-CSF has been proven having specific activity on neutrophil proliferation. However, improvement in the rhG-CSF preparation is still needed and longer administration of the rhG-CSF has to be applied in the future study.Keywords: rhG-CSF, biosimilar, neutropenia, Sprague Dawley rats

A phase I study of different doses and frequencies of pegylated recombinant human granulocyte-colony stimulating factor (PEG rhG-CSF) in patients with standard-dose chemotherapy-induced neutropenia

The recommended dose of prophylactic pegylated recombinant human granulocyte-colony stimulating factor (PEG rhG-CSF) is 100 μg/kg once per cycle for patients receiving intense-dose chemotherapy. However, few data are available on the proper dose for patients receiving less-intense chemotherapy. The aim of this phase I study is to explore the proper dose and administration schedule of PEG rhG-CSF for patients receiving standard-dose chemotherapy. Eligible patients received 3-cycle chemotherapy every 3 weeks. No PEG rhG-CSF was given in the first cycle. Patients experienced grade 3 or 4 neutropenia would then enter the cycle 2 and 3. In cycle 2, patients received a single subcutaneous injection of prophylactic PEG rhG-CSF on d 3, and received half-dose subcutaneous injection in cycle 3 on d 3 and d 5, respectively. Escalating doses (30, 60, 100 and 200 μg/kg) of PEG rhG-CSF were investigated. A total of 26 patients were enrolled and received chemotherapy, in which 24 and 18 patients entered cycle 2 and cycle 3 treatment, respectively. In cycle 2, the incidence of grade 3 or 4 neutropenia for patients receiving single-dose PEG rhG-CSF of 30, 60, 100 and 200 μg/kg was 66.67%, 33.33%, 22.22% and 0, respectively, with a median duration less than 1 (0-2) d. No grade 3 or higher neutropenia was noted in cycle 3 in all dose cohorts. The pharmacokinetic and pharmacodynamic profiles of PEG rhG-CSF used in cancer patients were similar to those reported, as well as the safety. Double half dose administration model showed better efficacy result than a single dose model in terms of grade 3 neutropenia and above. The single dose of 60 μg/kg, 100 μg/kg and double half dose of 30 μg/kg were recommended to the phase II study, hoping to find a preferable method for neutropenia treatment.

Effect of recombinant human granulocyte colony-stimulating factor on Nogo receptor expression in the brain tissues of neonatal rats after hypoxic-ischemic brain damage

Objective To observe the different effects of 2 doses recombinant human granulocyte colony-stimulating factors(rhG-CSF)on Nogo receptor(NgR)expression in the brain tissue of neonatal rats after hypoxic-ischemic brain damage(HIBD)at different times in order to reveal the neuroprotective effects of rhG-CSF. Methods Seven-day neonatal Sprague-Dawley(SD)rats were randomly divided into 4 groups by drawing method: sham operation group, model group, low-dose rhG-CSF group and high-dose rhG-CSF group, 24 rats in each group.Then each group was divided into 4 subgroups(6 rats in each subgroup)and all rats were exterminated at different times after HIBD(1 d, 3 d, 7 d and 14 d). In the low-dose rhG-CSF group and high-dose rhG-CSF group, the rats were given daily doses of rhG-CSF 50 μg/kg, 100 μg/kg respectively for 7 days by subcutaneous injection immediately after the molding(total 7 injections). In model group, rats received an injection of same amount of 9 g/L saline.In sham operation group, rats received no special treatment.Brain tissues of rats from each group were collected at different time points.The expressions of NgR protein and NgR mRNA in the left brain tissue were detected by immunohistochemistry and real-time fluorescent quantitative polymerase chain reaction(PCR). Results Immunohistochemistry: NgR proteins were constitutively expressed in the cerebral cortex in sham operation group at each time point; compared with sham operation group, the expressions of NgR in model group were increased markedly at each time point (135.67±16.63, 173.98±17.82, 234.00±14.70, 319.59±25.22), and the differences were statistically significant(all P<0.01); compared with model group, the expressions of NgR in the cerebral cortex in low-dose rhG-CSF group (134.35±8.89, 109.04±12.62, 75.99±13.39)and high-dose rhG-CSF group (81.38±12.25, 80.14±10.50, 72.58±13.66) on the 3rd, 7th, 14th day were reduced significantly(all P<0.01). Compared with low-dose rhG-CSF group, the protein expressions of NgR in the high-does rhG-CSF group were decreased faster, and had the marked difference on the 3rd, 7th day (P<0.05). Real-time fluorescent quantitative PCR: compared with the sham operation group, the expressions of NgR mRNA increased gradually in the cerebral cortex in the model group (1.34±0.24, 1.88±0.27, 2.88±0.84, 4.26±0.86), the differences in NgR mRNA expression were statistically significant at different times(all P<0.05); compared with model group, the expressions of NgR mRNA in low-dose rhG-CSF group on the 7th (1.08±0.30), 14th day (0.93±0.26) and high-dose rhG-CSF group on the 3rd(0.61±0.10), 7th(0.56±0.28), 14th day (0.47±0.12) were significantly different(all P<0.05). The expressions of low-dose group and high-dose group were reduced gradually.The NgR mRNA expression reduced more quickly in the high-dose group than in the low-dose rhG-CSF group and had substantial difference between two groups in 3 days (P<0.05). Conclusions The findings suggest that rhG-CSF intervention can reduce the expressions of NgR in the brain tissues of neonatal rats after HIBD, and low-dose rhG-CSF also has neuroprotective effect, but it could be weaker than high-dose rhG-CSF. Key words: Recombinant human granulocyte colony-stimulating factor; Hypoxic ischemic brain damage; Nogo receptor

Recombinant Human Granulocyte Colony-Stimulating Factor Promotes Preinvasive and Invasive Estrogen Receptor-Positive Tumor Development in MMTV-erbB2 Mice.

PurposeWe investigated whether recombinant human granulocyte colony-stimulating factor (rhG-CSF) could promote the development of preinvasive and invasive breast cancer in mouse mammary tumor virus (MMTV-erbB2) mice with estrogen receptor-positive tumors.MethodsMMTV-erbB2 mice were randomly divided into three experimental groups with 20 mice in each group. MMTV-erbB2 mice were treated with daily subcutaneous injections of vehicle or rhG-CSF (low-rhG-CSF group, rhG-CSF 0.125 µg; vehicle-rhG-CSF group, normal saline 0.25 µg; and high-rhG-CSF group, rhG-CSF 0.25 µg) at 3 months of age. Cellular and molecular mechanisms of G-CSF action in mammary glands were investigated via immunohistochemistry and reverse transcription polymerase chain reaction.ResultsLow, but not high, rhG-CSF doses significantly accelerated mammary tumorigenesis in MMTV-erbB2 mice. Short-term treatment with rhG-CSF could significantly promote the development of preinvasive mammary lesions. The cancer prevention effect was associated with reduced expression of proliferating cell nuclear antigen, cluster of differentiation 34, and signal transducers and activators of transcription 3 in mammary glands by >80%.ConclusionWe found that G-CSF was regulated by rhG-CSF both in vitro and in vivo. Identification of G-CSF genes helped us further understand the mechanism by which G-CSF promotes cancer. Low doses of rhG-CSF could significantly increase tumor latency and increase tumor multiplicity and burden. Moreover, rhG-CSF effectively promotes development of both malignant and premalignant mammary lesions in MMTV-erbB2 mice.

Physiologically-Based Mathematical Modelling of Neutrophil Dynamics during Concurrent Chemotherapy and Filgrastim Support

A common and dose-limiting side effect of chemotherapy is the development of neutropenia (a reduction in neutrophil numbers). To avoid or mitigate drops in absolute neutrophil counts (ANCs), patients are typically given recombinant human granulocyte colony-stimulating factor (rhG-CSF/filgrastim) to minimise the myelosuppressive nature of anti-cancer treatments. Dosing recommendations for filgrastim after chemotherapy suggest treatment begin one day post-chemotherapy and continue for a given amount of time or until ANCs rise sufficiently. Indeed, filgrastim support in a given chemotherapy cycle can sometimes reach seven to ten days in a 14-day period. Due to the intricacy of neutrophil production from the hematopoeitic stem cells in addition to the complexity of the interactions of cytokines and their receptors, a complete understanding of the mechanisms underlying myelopoiesis remains elusive. Mathematical modelling of these processes is a method which provides a global view of the dynamics of blood cell production and helps to elucidate the implications of concurrent chemotherapy and rhG-CSF support upon the blood production system. Moreover, the mathematical treatment of myelopoiesis can suggest novel dosing regimens that may be more beneficial than current schedules by supporting currently-held hypotheses and/or revealing previously unstudied relationships and dynamics.In this study, we construct a physiologically-based model of myelopoiesis which incorporates an up-to-date understanding of the production of neutrophils with our group’s previously published model of blood cell dynamics. This model is combined with pharmacokinetic and pharmcodynamic (PKPD) models of Zalypsis (PM00104), an anti-cancer drug currently in phase II clinical trials, and filgrastim, a myelostimulant. The physiological model of myelopoiesis directly relates observable delays in neutrophil production to temporal lags in the model through the use of delay differential equations. All parameters are comprehensively defined for an average patient by utilising previously published physiological and PKPD studies. The model is numerically implemented and simulated to compare its predictions to ANC time series of patients undergoing the CHOP14 protocol. Able to recreate previously published data, we then investigated the optimal timing of filgrastim administrations post-chemotherapy during 14-day periodic chemotherapy and examined the number of filgrastim administrations necessary to ward off neutropenia using this optimised timing.Our results indicate that delaying rhG-CSF administrations by six or seven days after the administration of chemotherapy lessens the myelosuppressive impact of anti-cancer treatment. In addition, we found that if filgrastim administration are started seven days post-chemotherapy, as few as three or four doses of rhG-CSF during a 14-day cycle would improve the ANC nadir experienced by an average patient during myelosuppressive chemotherapy. In all, our results suggest that it is possible to lessen the hematopoietic burden of chemotherapy on patients and that detailed physiological modelling of myelopoiesis is a useful tool to clinicians and researchers alike. DisclosuresOff Label Use: We look at optimal dosing regimens of filgrastim during periodic chemotherapy in the context of physiological mathematical models. No clinical trials were undertaken and no patients underwent any regimen changes..

The effect of gradual increment in rhG-CSF dose on stem cell yields in patients with multiple myeloma mobilized with intermediate dose cyclophosphamide plus rhG-CSF

Cyclophosphamide along with recombinant human granulocyte-colony stimulating factor (rhG-CSF) is a commonly used strategy for mobilization. However, the optimal timing for rhG-CSF initiation after cyclophosphamide has not been determined as conclusively as has the G-CSF dose. In this paper, we aimed to present gradual dose increment of rhG-CSF between the third day of mobilization and time to apheresis that is started with 5 μg/kg (from day 3 to day 7) and continued with 10 μg/kg (from day 8 to time to apheresis) for peripheral blood stem cell (PBSC) mobilization in multiple myeloma (MM) patients and its effect on stem cell yield and mobilization success. Data from 30 consecutive patients with MM who underwent PBSC mobilization between October 2011 and June 2013, were retrospectively reviewed. While twenty-eight of 30 patients (93.3%) were successfully mobilized, 2 patients (6.7%) had mobilization failure. The final median CD34+ cell dose harvested from the patients was 9.5×10(6)/kg. The median number of apheresis was 2.5 (range, 0-3). Twenty-four patients (80%) yielded >2×10(6) CD34+ cells/kg in one apheresis procedure. In conclusion, our regimen might be used to decrease the mobilization failure regarding the low dose rhG-CSF use and provide a cost effective strategy.

Successful haploidentical PBSCT with subsequent T‐cell addbacks in a boy with HyperIgM syndrome presenting as severe congenital neutropenia

HIGM syndrome is a group of primary immunodeficiency disorders characterized by recurrent bacterial and opportunistic infections; it is also associated with normal to elevated serum IgM levels and a concomitant deficiency of IgG, IgA, and IgE. In this report, we give account of a boy with X-linked HIGM and a novel Y172C mutation within his CD40LG gene. He presented with severe neutropenia as the dominating symptom. His bone marrow showed maturation arrest at the promyelocyte/myelocyte stage, typical of congenital neutropenia. This boy suffered from life-threatening infections and required high doses of rhG-CSF, and a haploidentical PBSCT was also successfully performed, thus leading to reconstitution of CD40L expression on activated CD4+ T cells (as assessed with flow cytometry six months after the procedure). Two low-dose T-cell addbacks were required to re-establish full donor chimerism and clear CMV reactivation. The report demonstrates that in select cases, alternative donor allogeneic HSCT supported by DLI may be effective in correcting the defect in X-linked HIGM, and HSCT in HIGM children is not necessarily limited to matched sibling donor transplantation.

Mobilization of PBSCs with chemotherapy and recombinant human G-CSF: a randomized evaluation of early vs late administration of recombinant human G-CSF

The optimal timing for recombinant human (rh)G-CSF administration after chemotherapy for PBSC mobilization has not yet been determined. In this study, we compared two different time schedules of rhG-CSF; 4th (early) vs 7th day (late), in 48 consecutive patients with multiple myeloma and lymphoma undergoing PBSC mobilization with CE (CY 4 g/m(2) on day 1 and etoposide 200 mg/m(2) on days 1-3). The rhG-CSF dose was 10 microg/kg/day for all patients. Both groups were comparable in terms of sex, age and number of previously given different chemotherapy regimens. Duration of neutropenia, CD34(+) cell count on the first day of apheresis and numbers of aphereses were not statistically different between the two arms. However, the number of doses of rhG-CSF up to the first cycle of apheresis procedures was significantly lower in the late group than in the early group (P=0.005). The median number of total CD34(+) cells collected was 10.54 x 10(6)/kg (range 0.11-37.27) in the early group and 10.81 x 10(6)/kg (range 0.17-49.83) in the late group of rhG-CSF (P=0.781). We conclude that PBSC mobilization after late use of rhG-CSF is an effective approach and therefore, in routine clinical practice, late rhG-CSF may be used for PBSC collections after chemotherapy-based mobilization regimens in this cost-conscious era.

Recombinant human G-CSF increases invasiveness of breast cancer cells.

Abstract Abstract #4057 Background: Recombinant human granulocyte-colony stimulating factor (rhG-CSF) is commonly given to breast cancer patients to prevent neutropenia during chemotherapy. It binds to the G-CSF receptor (G-CSFR) on neutrophil precursors, promoting their proliferation within the bone marrow. We observed breast cancer growth in patients with advanced breast cancer during rhG-CSF administration. The G-CSFR is present on some solid tumors, but has not been examined in breast cancer. We asked whether breast cancer cells express the G-CSFR, and whether rhG-CSF has any direct effects on breast cancer proliferation or invasion in vitro.&amp;#x2028; Material and Methods: MDA MB231 is a highly aggressive ER–PR–Her2/neu– breast cancer cell line derived from a human pleural effusion. We analyzed this cell line for the presence of the G-CSFR by immunohistochemistry, reverse-transcriptase PCR (RT-PCR), Western Blot analysis and flow cytometry. We sequenced DNA products of RT-PCR to verify results. We examined the effects of physiologic doses of rhG-CSF on breast cancer cell proliferation and invasion using MTT proliferation and Matrigel transwell assays, respectively.&amp;#x2028; Results: MDA MB231 breast cancer cells express G-CSFR protein and mRNA. Of seven known G-CSFR isoforms, we found evidence of isoforms 1 and 4. Treating MDA MB231 breast cancer cells with rhG-CSF at 50ng/ml did not increase cell proliferation, but did increase invasion across a Matrigel-coated transwell membrane. Cells treated with rhG-CSF appeared more stellate than untreated cells. Studies are ongoing to evaluate which signal transduction pathways and genes are involved in this increased invasion response. The effects of rhG-CSF on breast cancer growth in vivo are also being examined in murine models.&amp;#x2028; Discussion: rhG-CSF is given routinely as part of many adjuvant chemotherapy regimens for breast cancer. Although dose-dense regimens using rhG-CSF have shown equivalent or improved survival when compared to standard dosing regimens, it is possible that rhG-CSF may increase the growth or invasive potential of some breast cancers. It is also possible that rhG-CSF could mobilize breast cancer cells from the bone marrow, making these cells more susceptible to chemotherapy. These possibilities should promote thoughful use of rhG-CSF in the clinical setting, and should factor into the design and implementation of clinical trials using rhG-CSF. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 4057.

PO2-43 PROBUCOL DOSE-DEPENDENTLY PROMOTES REVERSE TRANSPORT OF CHOLESTEROL FROM MACROPHAGES TO FECES IN VIVO

Background and objectives: Donor lymphocyte infusions (DLI) have become widely used for prevention or treatment of relapse after allogeneic hematopoietic stem cell transplantation. Increasing use of reduced intensity conditioning regimens (RICR) and subsequent application of DLI forced the hemapheresis centers to collect donor lymphocytes in certain quantity and quality. The place of growth factors especially granulocyte colony stimulating factor (rhG-CSF, filgrastim) in allogeneic hemapoietic stem cell (HSC) collection is established, but there is no consensus about the role of rhG-CSF. We aimed to clarify the dose effect of rhG-CSF on lymphocyte subpopulations (CD3+, CD3+4+, CD3+8+, CD19+, CD3−16+56+) cells and CD34+ HSC.Donors and methods: Major indications for DLI (mean volume: 180 ± 52 ml) were for relapse or transplants using RICR mainly in patients with acute leukemia (n=20) or chronic myeloid leukemia (n=15). In four years we performed 40 lymphocyte apheresis (LA) on 30 healthy (med. age 28, M/F 21/9) donors using continuous flow cell separators by processing 2–2.5 times of their total blood volume (TBV). The apheresis data is divided into three groups according to rhG-CSF dose used for priming. Donors in Group I (n=18), Group II (n=9) and Group III (n=13) received no rhG-CSF (steady state), rhG-CSF 5 μg/kg/d sc × 5 days and rhG-CSF 10 μg/kg/d sc × 5 days, respectively. There was no difference within groups concerning TBV processed and recipient body weight.Results: A total of 11,565 ml (±3700) of blood was processed in 216 min (±36.5) at an inlet of 56.8 ml/min (±10.6) using 999 ml (±307) ACD. The CD34+ HSC increased with increasing rhG-CSF dose as expected. Median CD3+ lymphocyte yield per recipient body weight in Group I, II and III were 0.9 × 10e8/kg (range: 0.1–2.1), 2.9 × 10e8/kg (range: 1.6–4.3) and 2.1 × 10e8/kg (range: 0.6–6.9), respectively. The primed donors T lymphocyte yield was 2–3-fold more in comparison to Group I. This gain was most significant between Group I and III in terms of mean CD3+ (1.09 × 10e8/kg vs 2.41 × 10e8/kg, p=0.02), CD3+4+ (0.64 × 10e8/kg vs 1.44 × 10e8/kg, p=0.02) and CD3+8+ (0.42 × 10e8/kg vs 0.89 × 10e8/kg, p=0.03) cells, respectively.Conclusion: Though the yield of lymphocyte subsets in G-CSF primed donors exceeds the non-primed donors, the target range of 1 × 10e7–1 × 10e8/kg CD3+ lymphocytes could be achieved in the majority of the apheresis procedures without rhG-CSF priming. The yield of T and B lymphocyte subsets are increased by G-CSF stimulation but not on a logarithmic scale, which did not correlate into a clinical relevance.

Therapeutic Angiogenesis With Intramuscular Injection of Low-Dose Recombinant Granulocyte-Colony Stimulating Factor

In vivo administration of granulocyte colony-stimulating factor (G-CSF) has been shown to facilitate regeneration of cardiovascular tissues. However, G-CSF causes marked leukocytosis that potentially induces adverse cardiovascular events. Earlier studies showed that G-CSF had direct stimulatory actions on mature endothelial cells, resulting in promotion of angiogenesis. We thus examined whether low doses of recombinant human G-CSF (rhG-CSF) locally injected into ischemic tissues would stimulate angiogenesis without inducing severe leukocytosis. Reverse-transcription polymerase chain reaction (PCR) revealed expression of G-CSF receptor in human umbilical vein endothelial cells (HUVECs). rhG-CSF (100 ng/mL) enhanced migration and tube formation but not proliferation of HUVECs in vitro. We then examined the effects of rhG-CSF on angiogenesis in a rat model of hindlimb ischemia. Nude rats received in their ischemic skeletal muscles either rhG-CSF (2, 10, 20 microg/kg per day) or saline (control) for 6 days. Laser Doppler blood flowmetry (LDBF) revealed an augmented ischemic/normal limb LDBF ratio and an increased capillary density in the rhG-CSF-treated groups compared with the control at days 14, 21, and 28 (P<0.05). These doses of rhG-CSF induced only mild leukocytosis ( approximately 1.4-fold increases versus baseline). rhG-CSF promoted endothelial migration and tube formation in vitro. Local injection of low doses rhG-CSF effectively augmented ischemia-induced angiogenesis in vivo. This treatment regimen of low-dose rhG-CSF may become a new and safe modality for therapeutic angiogenesis.

Significance of donors treatment with combination of recombinant human interleukin-11 and recombinant human granulocyte-colony stimulating factor in reduction of incidence of lethal grafts versus host disease after allogeneic bone marrow transplantation

To investigate the effects of donor treatment with combination of recombinant human interleukin-11 (rhIL-11) and recombinant human granulocyte-colony stimulating factor (rhG-CSF) on incidence of lethal graft versus host disease (GVHD) after allogeneic bone marrow transplantation (BMT). Forty C57BL/6 mice, as donors, were treated with phosphate buffered saline (PBS), or optimal doses of rhG-CSF, rhIL-11, or rhG-CSF + rhIL-11 for five consecutive days. Fifty BALB/C mice, as recipients, received a lethal dose of 8.0 Gy total body irradiation (TBI) 4-6 h before the transplantation. Ten C57BL/6 mice without transplantation were used as pure radiation control group (group A). The other 40 C57BL/6 mice received the allogeneic bone marrow from the donors treated with PBS (group B), rhG-CSF (group C), rhIL-11 (group D), or rhG-CSF + rgIL-11 (group E) 14 and 30 days after the transplantation, suspensions of bone marrow cells were prepared from the 4 groups. Monoclone antibody of PE-H-2D(b) was added. The percentage of the H-2D(b) positive cells was observed by flow cytometry to detect the chimerism to determine the proportion of the cells of donor origin. The liver, spleen, intestine, and skin tissues of the mice presenting symptoms of GVHD were taken out before the diseased mice died. The long-surviving mice were killed 60 days after the transplantation and the above-mentioned tissues were taken out. Light microscopy was used to observe the pathological changes. 14 days after transplantation, the spleen cells of the different groups were obtained to undergo mixed leucocyte reaction (MLR), and 14 days after transplantation another spleen cells of groups B, C, D, and E were obtained to prepare suspension of mononuclear cells to test the levels of IL-4, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha by ELISA. The survival time of group E was 38 d +/- 19 d, significantly longer than the other groups (all P < 0.01). The survival times of groups C and D were 23 d +/- 8 d and 25 d +/- 15 d respectively, both significantly longer than that of group B (14 d +/- 4 d, both P < 0.01). The mice of group A all died within 8 days with a survival time of 6.2 d +/- 1.3 d. The GVHD pathological changes were milder in group E than in the other groups. 14 and 30 days after transplantation, the percentage of H-2D(b) antibody positive cells was over 95% in all surviving transplanted mice. All the mice of group B developed GVHD, and the earlier GVHD appeared the severer the pathological changes. In group E 3 mice developed GVHD more than 20 days after transplantation, 7 mice survived more than 30 days, and 4 survived 60 days without any sign of GVHD. The pathological changes in the tissues of the mice that died within 20 days were more remarkable than those of the mice that died 20 days or more after transplantation. The levels of IFN-gamma of group C, D, and E were significantly lower than that of group B, and the levels of IL-4 of group C, D, and E were significantly higher than that of group B (all P < 0.01). The level of IFN-gamma of group E was significantly lower than those of groups C and D, and the level of IL-4 of group E was significantly higher than those of groups C and D (all P < 0.05). The levels of group D and E were significantly lower than those of group B and C (all P < 0.01). The MLR 14 days after transplantation showed that the proliferation to host alloantigen of group E was 0.22 +/- 0.08, significantly lower than those of groups B, C, and D (0.87 +/- 0.14, 0.54 +/- 0.21, and 0.49 +/- 0.18 respectively, all P < 0.01). Donor treatment with the combination of rhIL-11 and rhG-CSF shows a synergic function in inducing immune tolerance, and significantly reduces the incidence of lethal GVHD after BMT.

Clinical study on diyushengbai tablet to prevent myelosuppression of patients with breast cancer by neoadjuvant chemotherapy

目的 观察地榆升白片对乳腺癌新辅助化疗后外周血象的影响.方法69例乳腺癌患者分为观察组和对照组,两组术前接受CAF方案化疗2个疗程,化疗后均给予重组人粒细胞集落刺激因子(G-CSF,商品名:瑞白)治疗.末次化疗后2～3周患者血象恢复正常后接受乳腺癌改良根治术.观察组加服地榆升白片.观察两组外周血象变化及瑞白用量等指标.结果观察组Ⅲ度及Ⅳ度骨髓抑制发生率12.5%,显著低于对照组的40.5%(x2=3.95,P 0.05),对照组外周血WBC、PLT、Hb治疗后较治疗前明显下降,其中PLT差异有显著意义(P<0.05);观察组、对照组人均瑞白(100 μg)用量分别为(2.12±0.64)支和(2.52±0.86)支,差异有显著意义(f=2.16,P<0.05).结论地榆升白片可预防乳腺癌化疗的骨髓抑制作用,提高PLT水平,降低WBC下降幅度,减少升白药的用量,值得推广。

RhG-CSF does not affect the phenotype of adult donor peripheral blood NK cells

Considerable evidence in preclinical models as well as in human transplantation now suggests that donor-derived natural killer (NK) cells can contribute to alloimmune recognition of recipient residual tumour cells. This makes the NK cell population an attractive target for in vitro or in vivo manipulations, in order to improve the antitumour effect of allogeneic transplantation. However, conditions in which allogeneic donor cells are collected vary; several reports have emphasised the different phenotypic and functional properties of T cells derived from marrow, cord blood or mobilised peripheral blood grafts; others have demonstrated different clinical outcomes following blood or marrow transplantation after myeloablative conditioning regimens. NK cells have been examined in this setting; the availability of new tools to study the expression of a variety of surface antigens that are involved in the control of NK cell activity offered us an opportunity to extensively characterise the phenotypic properties of NK cells from donors, before and after administration of pharmacological doses of rhG-CSF used for haematopoietic progenitor mobilisation. Our study suggests that rhG-CSF does not reproducibly alter blood NK cell phenotype in normal individuals, and thus that donor-derived cells are fully equipped to exert their potential antitumour effect.