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PubMed2006

Peripheral blood mobilization of different lymphocyte and dendritic cell subsets with the use of intermediate doses of G-CSF in patients with non-Hodgkin's lymphoma and multiple myeloma.

Between June 2003 and November 2004, we collected mobilized peripheral blood units from 29 patients with non-Hodgkin's lymphoma and multiple myeloma for autologous peripheral blood stem cell transplantation. They received granulocyte colony-stimulating factor (G-CSF) (16 micro g/kg/day) for a total of 5 days. Immediately before and 3 h after the fourth and fifth dose of G-CSF, we performed flow cytometry analysis to quantify: T cells (CD3+CD4+, CD3+CD8+), B cells (CD19+), NK cells (CD3-CD16+CD56+), NKT cells (CD3+CD16+CD56+), type 1 dendritic cells (DC1) (lin-HLA-DR+CD11c+), type 2 dendritic cells (DC2) (lin-HLA-DR+CD123+), regulatory T cells (Tregs) (CD4+CD25+), and activated T cells (CD3+HLA-DR+). All cell subsets were mobilized after G-CSF treatment with the exception of B, NK, and NKT lymphocytes. The median number of Treg cells before and after G-CSF was statistically different (29+/-14.9x10(6)/l vs 70.1+/-46.1x10(6)/l, P<0.02). DCs were mobilized significantly with a 5.9-fold increase in DC2 (15.1+/-30.3x10(6)/l vs 89.8+/-81.0x10(6)/l, P<0.02) and a 2.6-fold increase for DC1 (41+/-42.5x10(6)/l vs 109.5+/-58.0x10(6)/l, P<0.04). Patients received a mean of 3.1+/-1.2x10(7)/kg NK cells, 1.3+/-0.9x10(7)/kg NKT cells, 0.41+/-0.29x10(7)/kg DC1, 0.2+/-0.22x10(7)/kg DC2, and 1.8+/-1.9x10(7)/kg Tregs. In conclusion, intermediate doses of G-CSF induce mobilization of different lymphocyte subsets, with the exception of B, NK, and NKT cells. The mobilization of certain suppressive populations (DC2 and Treg) could be in theory deleterious, at least in patients with cancer.

Adult↗
PubMed2003

Synergistic effects of co-administration of angiotensin 1-7 and Neupogen on hematopoietic recovery in mice.

PURPOSE: Angiotensin 1-7 [A(1-7)] is a seven amino acid peptide that has been shown to increase the proliferation of epidermal stem cells after dermal injury and the number of hematopoietic progenitors in the bone marrow of myelosuppressed mice. In this study, the effect of combining A(1-7) with Neupogen on hematopoietic recovery and bone marrow progenitors was evaluated. MATERIALS AND METHODS: Intravenous 5-fluorouracil (5FU) was administered to induce myelosuppression. Administration of A(1-7) and/or Neupogen was initiated 2 days after chemotherapy. Angiotensin II (AII) and A(1-7) binding were assessed by flow cytometric analysis. Hematopoietic progenitors were counted by colony forming assays. Recovery of formed elements in the blood was evaluated by hemocytometer. RESULTS: Flow cytometric analysis indicated that the number of early hematopoietic progenitors (Lin(-)Sca1(+)cKit(+)) that bind AII or A(1-7) increased 5-7 days after intravenous injection of 150 mg/kg 5FU. Further, administration of A(1-7) led to a slight increase in the number of circulating leukocytes and platelets after this chemotherapeutic regimen. When given in combination with a subclinical dose of Neupogen, a synergistic effect on the number of circulating leukocytes was observed, but there was no further effect on the number of circulating platelets. In myelosuppressed mice, A(1-7) had its most profound effect on the number of hematopoietic progenitors in the bone marrow. The progenitors evaluated in the study included BFU-E, CFU-Meg, CFU-GM and CFU-GEMM. There was an increase in the number of these progenitors in the bone marrow, indicating an effect on all hematopoietic lineages. When given in combination with Neupogen, these effects were synergistic for the numbers of BFU-E and CFU-Meg (Neupogen by itself had no effect) and for the myeloid progenitors at lower doses of A(1-7). CONCLUSIONS: These results suggest that these hematopoietic agents act at different sites within the hematopoietic cascade and that combining these two agents may be of benefit in the treatment of hematopoietic disorders.

Angiotensin I↗
PubMed1997

Parvovirus B19 associated neutropenia. Treatment with Rh G-CSF.

In a 39-year-old patient under follow-up for metastatic adenocarcinoma of the colon, a post-operative fever and a profound neutropenia occurred during hospitalization. A diagnosis of primary infection with parvovirus B-19 was made in view of the association of reticulocytopenia, together with anti-B19 IgM antibodies and positive serum results for viral DNA. The granulocytic lineage appeared to be normal on bone-marrow smears, and anti PMN autoantibodies were not found. Rh G-CSF therapy resulted in a sharp increase in the PMN count. Although the pathophysiology of B19-associated neutropenia remains unclear, Rh G-CSF seems to be effective in this disorder.

Adenocarcinoma↗
PubMed2005

Myelodysplastic syndrome associated with multiple autoimmune disorders.

The association between myelodysplastic syndromes (MDS) and autoimmune manifestations is not uncommon. As a rule, autoimmune abnormalities follow the diagnosis of MDS. We describe here a patient with MDS who developed a striking spectrum of diverse autoimmune disorders, including dermatitis, polyarthritis, and vasculitis, which preceded the clinical appearance of MDS.

Aged↗
PubMed2005

Peculiarities of proliferation of epidermal cambial cells in mouse skin.

The long axis of epidermal cambial cells determining the direction of their division is determined before the beginning of division and is oriented perpendicularly, but not parallel, to the basal membrane, as in other basal cells. As a result, only one of the two newly formed cells adheres to the basal membrane and at the expense of traction forces detaching one cell from the other and elastic force of the basal membrane is formed as a maternal cell and stays in the focus of multiplication. The other cell, turning around under the effect of traction forces perpendicularly to the maternal cell, cardinally changes the direction of its long axis and is polarized parallel to the basal membrane like other cells. This cell becomes the daughter cell, its shape with the "head" and "tail" allows its rapid migration into other rosettes for differentiation.

Animals↗
PubMed2005

Optimization and applications of CDAP labeling for the assignment of cysteines.

PURPOSE: The aim of the study is to provide a methodology for assigning unpaired cysteine residues in proteins formulated in a variety of different conditions to identify structural heterogeneity as a potential cause for protein degradation. METHODS: 1-Cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) was employed for cyanylating free cysteines in proteins and peptides. Subsequent basic cleavage of the peptide bond at the N-terminal side of the cyanylated cysteines provided direct information about their location. RESULTS: CDAP was successfully employed to a wide variety of labeling conditions. CDAP was reactive between pH 2.0 and 8.0 with a maximum labeling efficiency at pH 5.0. Its reactivity was not affected by excipients, salt or denaturant. Storing CDAP in an organic solvent increased its intrinsic stability. It was demonstrated that CDAP can be employed as a thiol-directed probe to investigate structural heterogeneity of proteins by examining the accessibility of unpaired cysteine residues. CONCLUSION: CDAP is a unique cysteine-labeling reagent because it is reactive under acidic conditions. This provides an advantage over other sulfhydryl labeling reagents as it avoids potential thiol-disulfide exchange. Optimization of the cyanylation reaction allowed the utilization of CDAP as a thiol-directed probe to investigate accessibility of sulfhydryl groups in proteins under various formulation conditions to monitor structural heterogeneity.

Affinity Labels↗
PubMed1996

r-metHuG-CSF support to ifosfamide, cisplatin, etoposide chemotherapy in non-small cell lung cancer.

Twenty patients with locally advanced or metastatic non-small cell lung cancer entered a study of recombinant human methionyl G-CSF (r-metHuG-CSF) as an adjunct to ifosfamide, cisplatin and etoposide (IPE) regimen. Chemotherapy consisted of three courses of cisplatin 25 mg/m2, ifosfamide 1.5 g/m2 (with uroprotection) and etoposide 100 mg/m2 given on days 1-4 of a 21-day cycle. r-metHuG-CSF, 5 micrograms/kg, was administered subcutaneously from day 5 to day 14. Eighteen out of 20 patients completed the three courses (57 evaluable cycles). Grade 3-4 neutropenia affected 50, 42 and 22% of the patients during cycles 1, 2 and 3, respectively, whereas thrombocytopenia was observed in 25% of the patients throughout the chemotherapy protocol. Haematological toxic events requiring transfusions and/or antibiotics were responsible for 11 unplanned hospitalizations. Among these only three were exclusively devoted to febrile neutropenia care, the remaining eight being mainly required for blood transfusions. There were no deaths during the study duration. Dose reductions were needed in 65% of the patients and chemotherapy was delayed by thrombocytopenia in five patients. The total relative dose intensity was 84%. Eleven (55%) patients responded (one complete and 10 partial responses). Median survival was 9.5 months. We concluded that IPE combination chemotherapy can be administered safely with the support of r-metHuG-CSF inasmuch as neutropenia appears as mild to moderate and manageable. Optimal delivery of chemotherapy is still limited by other toxicities, mainly thrombocytopenia, but the successful relative dose intensity observed herein deserves further studies designed to analyze a dose intensity-survival relationship in non-small cell lung cancer.

Aged↗
PubMed1996

Effects of granulocyte colony-stimulating factor on the hemostatic system in healthy volunteers.

We previously identified a receptor for granulocyte colony-stimulating factor (G-CSFR) on platelet membranes, and reported that G-CSF enhanced ADP-induced platelet aggregation. Here, we investigated the priming effect of G-CSF on the hemostatic system in healthy volunteers given G-CSF. Following the administration of rhG-CSF (10 micrograms/kg for 30 min div) to 10 healthy volunteers, we found a significant elevation in the maximum platelet aggregation rate induced by ADP or collagen, thromboxane B2 level and amount of thrombin-antithrombin III complex. The D-D dimer and plasminogen activator inhibitor-1 showed no significant changes. These observations indicate that G-CSF administration may induce hypercoagulability in susceptible subjects. Therefore, patients or donors at risk of thrombosis or hypercoagulable state should be followed carefully after G-CSF administration.

Adenosine Diphosphate↗
PubMed2006

Granulocyte growth factors in the treatment of non-small cell lung cancer (NSCLC).

Neutropenia and subsequent infections are common events that limit treatment of non-small cell lung cancer (NSCLC). Granulocyte growth factors (G- and GM-CSF) have been introduced in clinical practice and their use has yielded a reduction of the infection risk related to chemotherapy and a dose increase of drug delivery. Randomized clinical trials have shown that granulocyte colony-stimulating factors and, more recently, the longer-acting pegylated granulocyte colony-stimulating factor (pegfilgrastim) effectively reduce the incidence and severity of neutropenia and of its complications. Recommendations for the use of haematopoietic colony-stimulating factors from the American Society of Clinical Oncology (ASCO) have been published in 1994 and updated in 1996, 1997 and 2000. Recently, moreover, National Comprehensive Cancer Network (NCCN) guidelines for the myeloid growth factors in cancer treatment make available. Chemotherapy-associated myelosuppression is a major limitation of anticancer therapy also in early stage, local advanced and metastatic NSCLC. Recently, dose-dense chemotherapy has been shown to improve the outcome in early stage breast cancer and non-Hodgkin's lymphoma. However, few randomized trials have been reported on chemotherapy with or without granulocyte growth factors as primary prophylaxis in NSCLC. Presently, there is no evidence for a benefit in response rate and survival from the use of granulocyte growth factors as support of chemotherapy, in particular, for locally advanced and metastatic NSCLC. In clinical practice, the role of granulocyte growth factors for NSCLC treatment should be limited following the guidelines. An appropriate use of granulocyte growth factors may reduce the overall cost of treatment and improve the quality of life, important aims in the treatment of patients with local advanced or metastatic NSCLC. In the future, we need to identify patients who can benefit from granulocyte growth factors for optimize the schedule and doses, in advanced disease and also, after the recent positive results of adjuvant chemotherapy, in early stages. This review summarizes the present knowledge on the use of granulocyte growth factors in NSCLC.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed2006

Safety and efficacy of once-per-cycle pegfilgrastim in support of ABVD chemotherapy in patients with Hodgkin lymphoma.

The objective of this prospective study was to examine the safety and efficacy of pegfilgrastim in support of chemotherapy regimens that are administered every 14 d. Patients with Hodgkin lymphoma receiving standard ABVD (doxorubicin, bleomycin, vinblastine, and decarbazine) chemotherapy every 14 d were eligible for the study. All patients received one fixed dose of 6 mg pegfilgrastim approximately 24h after each ABVD infusion. Absolute neutrophil counts (ANCs) were measured weekly while on therapy. After completion of therapy, ANCs were measured every 3-4 months for 2 years, and every 6 months thereafter. Twenty-three patients received 115 courses (230 doses) of ABVD. Nadir ANC below 0.5 x 10(9)/L was observed only in two cases (1.1%), and ANC counts on the day of ABVD therapy below 1 x 10(9)/L was observed only once (0.4%). No neutropenic fever was observed. Two hundred and fifteen (93.5%) doses of ABVD were given on time as scheduled. Grade 3/4 bone pain was observed after 0.4% of ABVD doses, and bleomycin lung toxicity was observed in two patients. With a median follow-up of 29 months after completion of ABVD, no myelodysplastic syndrome was observed. Two patients relapsed and subsequently underwent successful stem cell collection and transplantation. Pegfilgrastim was found to be safe in support of ABVD chemotherapy that is administered every 14 d. Pegfilgrastim was also effective in maintaining ABVD dose intensity, and keeping planned dose of chemotherapy on schedule.

Adult↗
PubMed2006

The effects of granulocyte colony-stimulating factor on the healing of tracheal anastomosis following radiation therapy in rats.

OBJECTIVE: This study evaluates the effects of granulocyte colony-stimulating factor on the healing of tracheal anastomosis following radiation therapy in rats. METHODS: Fifty-six male Wistar rats were divided into four groups. Group 1 underwent tracheal anastomosis. Group 2 underwent radiation therapy followed by tracheal anastomosis. Group 3 underwent radiation therapy followed by tracheal anastomosis and received granulocyte colony-stimulating factor. Group 4 underwent sham radiation therapy followed by sham tracheal anastomosis. At 10 days following radiation therapy, the trachea was dissected for histopathological, mechanical and biochemical evaluation. RESULTS: Median scores for inflammation were three points for Group 1, two points for Group 2, two points for Group 3 and one point for Group 4. Median scores for angiogenesis were four points for Group 1, two points for Group 2, three points for Group 3 and one point for Group 4. Median scores for connective tissue regeneration were four points for Group 1, two points for Group 2, three points for Group 3 and one point for Group 4. Median scores for epithelial regeneration were two points for Group 1, one point for Group 2, one point for Group 3 and one point for Group 4. Mean anastomotic bursting pressures were 853 mmHg for Group 1, 293 mmHg for Group 2, 417 mmHg for Group 3 and 966 mmHg for Group 4. Mean hydroxyproline concentrations were 159 microg/mg for Group 1, 177 microg/mg for Group 2, 120 microg/mg for Group 3 and 117 microg/mg for Group 4. CONCLUSIONS: This study suggests that granulocyte colony-stimulating factor contributes to the healing of tracheal anastomosis following radiation therapy through improved connective tissue regeneration.

Anastomosis, Surgical↗
PubMed2005

Proteomic signature of myeloproliferation and neutrophilia: analysis of serum and plasma from healthy subjects given granulocyte colony-stimulating factor.

OBJECTIVE: Proteomic analysis could improve our understanding of the mechanisms and consequences of myeloproliferation. Healthy subjects treated with granulocyte colony-stimulating factor (G-CSF) were used as a model of myeloproliferation. METHODS: Levels of 80 soluble factors were measured by enzyme-linked immunosorbent assay before and after 5 days of G-CSF. Both serum and plasma levels were measured to generate a comprehensive profile and determine whether serum or plasma best portrays biological and physiological changes. RESULTS: Comparison of samples collected prior to G-CSF demonstrated that 44 factors differed between serum and plasma. Concentrations of several growth factors and chemokines were greater in serum than in plasma, while the opposite was true for several interleukins. Following G-CSF serum levels of 14 factors and plasma levels of 15 factors changed. Eleven increased in both serum and plasma, including cell adhesion molecules (vascular cell adhesion molecule-1, E-selectin, and L-selectin), matrix metalloproteases (MMP-1, -8, and -13), cytokine receptors (tumor necrosis factor receptor 1 and 2, and interleukin-2 receptor), the acute phase reactant, serum amyloid A, and a growth factor, hepatocyte growth factor. CONCLUSION: Some protein levels differ markedly in serum and plasma. Myeloproliferation is associated with changes in the levels of several proteases, adhesion molecules, and cytokines.

Blood Proteins↗