OBJECTIVE: To investigate the influence of granulocyte colony stimulating factor (G-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF) on preimplantation development and implantation in mouse embryos. MATERIAL AND METHODS: Eight-cell stage mouse embryos were cultured for 96 hours with G-CSF or GM-CSF at concentrations of 10 pg/ml, 100 pg/ml, 1 ng/ml and 10 ng/ml. Embryos not treated with G-CSF or GM-CSF were served as control. The percentages of embryos which developed to expanded, hatched blastocyst stage and in vitro implantation at 96 hours were determined. Results were analyzed with Kolmogorov-Smirnov test and analysis of variance (ANOVA). The statistical significance was defined as p<0.05. RESULTS: The percentages of fully expanded blastocysts in all G-CSF and GM-CSF treatment groups were not significantly different from the control. The percentages of hatched blastocysts were significantly higher in 100 pg/ml and 10 ng/ml of G-CSF treatment group compared to the control (p<0.05, p<0.05, respectively). The percentages of hatched blastocysts were significantly lower in 1 ng/ml of GM-CSF treatment group compared to the control, 10 pg/ml, and 100 pg/ml of GM-CSF treatment group (p<0.05, p<0.05, p<0.05, respectively), and the percentages of hatched blastocysts were also significantly lower in 10 ng/ml of GM-CSF treatment group compared to the control and 100 pg/ml of GM-CSF treatment group (p<0.05, p<0.05, respectively). The percentages of implanted blastocysts in vitro were significantly higher following incubation with all concentrations of G-CSF compared to the control and, especially in 100 pg/ml and 10 ng/ml of G-CSF treatment groups compared to the control and other treatment groups. The percentages of implanted blastocysts in vitro were significantly higher in 10 pg/ml of GM-CSF treatment group than the control and 100 pg/ml of GM-CSF treatment groups (p<0.05, p<0.05, respectively). CONCLUSION: G-CSF and GM-CSF might influence on embryonic development and implantation in mouse embryos.
Animals ; Blastocyst ; Colony-Stimulating Factors* ; Embryonic Development ; Embryonic Structures* ; Female ; Granulocyte Colony-Stimulating Factor ; Granulocyte-Macrophage Colony-Stimulating Factor* ; Granulocytes* ; Mice* ; Pregnancy

Granulocyte macrophage-colony stimulating factor (GM-CSF) occupies a central position in the regulation of hematopoietic responses. GM-CSF not only signals proliferations of granulocyte-macrophage but also drives these cells into differentiation and activates mature cells of the GM-CSF sensitive lineage. Myelosuppression that is induced by M-VAC (methotrexate, vinblastine, doxorubicin, cisplatinum) chemotherapy brings many problems in successful treatment such as sepsis, dose reduction, delaying the schedule. Granulocyte-macrophage colony stimulating factor is introduced hopefully as a new solution for these problems. So we evaluated the efficacy and safety of GM-CSF in leukopenia induced by M-VAC chemotherapy in patients with urothelial cancer. GM-CSF was administered at 200ug subcutaneously in 10 M-VAC cycles of 6 patients on 5th and 6th day after M-VAC therapy. Sixteen cycles, by which only M-VAC chemotherapy was administered without GM-CSF. of the other 6 patients served as control group. Mean white blood cell count in peripheral blood at M-VAC 2nd day and 15th day was 5,630/mm3 and 4,240/mm3 in GM-CSF administered cycles, 6,58l/mm3and 3,613/mm3 in non GM-CSF administered cycles. There was no delayed cycle in administration of MTX and vinblastine at M-VAC 15th day in the cycles with GM-CSF. There was no significant side effects caused by GM- CSF. The result indicates that GM-CSF can be used safely and effectively against leukopenia after M-VAC chemotherapy of urothelial cancer.
Appointments and Schedules ; Colony-Stimulating Factors ; Doxorubicin ; Drug Therapy* ; Granulocyte-Macrophage Colony-Stimulating Factor* ; Granulocytes ; Humans ; Leukocyte Count ; Leukopenia* ; Sepsis ; Vinblastine

BACKGROUND: As an attempt to develop a strategy to improve the protective immune response to GM-CSF-secreting CT26 (GM-CSF/CT26) tumor vaccine, we have investigated whether the apoptogenic treatment of GM-CSF/CT26 prior to vaccination enhances the induction of anti-tumor immune response in mouse model. METHODS: A carcinogen- induced mouse colorectal tumor, CT26 was transfected with GM-CSF gene using a retroviral vector to generate GM-CSF-secreting CT26 (CT26/GM-CSF). The CT26/GM-CSF was treated with gamma-irradiation or mitomycin C to induce apoptosis and vaccinated into BALB/c mice. After 7 days, the mice were injected with a lethal dose of challenge live CT26 cells to examine the protective effect of tumor vaccination in vivo. RESULTS: Although both apoptotic and necrotic CT26/GM-CSF vaccines were able to enhance anti-tumor immune response, apoptotic CT26/GM-CSF induced by pretreatment with gamma-irradiation (50,000 rads) was the most potent in generating the anti-tumor immunity, and thus 100% of mice vaccinated with the apoptotic cells remained tumor free for more than 60 days after tumor challenge. CONCLUSION: Apoptogenic pretreatment of GM-CSF-secreting CT26 tumor vaccine by gamma-irradiation (50,000 rads) resulted in a significant enhancement in inducing the protective anti-tumor immunity. A rapid induction of apoptosis of CT26/GM-CSF tumor vaccine at the vaccine site might be critical for the enhancement in anti-tumor immune response to tumor vaccine.
Animals ; Apoptosis ; Colony-Stimulating Factors* ; Colorectal Neoplasms ; Granulocyte-Macrophage Colony-Stimulating Factor ; Mice ; Mitomycin ; Vaccination ; Vaccines ; Zidovudine

Nutrigenomics refers to research that investigates the interaction between nutrition and the human genome. Caffeine in tea and coffee is widely and routinely consumed by people. This study was performed to confirm the effect of caffeine treatment on the gene expression and cytokine profiling in 3T3-L1 adipocyte cells using microarray and protein array methodology. Treatment of caffeine in 3T3-L1 adipocyte cells increased expression of several genes related with obesity including adipocyte C1Q and collagen domain containing (ACDC), Adipsin (ADN), uncoupling protein 3 (UCP3), while glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which is known as lipid storage enzyme, was decreased by caffeine treatment. Furthermore, cytokines, such as interleukin-3 (IL-3), interleukin-12 (IL-12), interleukin-13 (IL-13), granulocyte colony stimulating factor (GCSF), granulocyte macrophage colony stimulating factor (GM-CSF) and vascular endothelial growth factor (VEGF), were decreased in caffeine treated 3T3-L1 adipocyte cells. These results provided interesting information about the genes related with caffeine and cytokine expression profiling in obesity.
Adipocytes* ; Caffeine* ; Coffee ; Collagen ; Colony-Stimulating Factors ; Complement Factor D ; Cytokines ; Gene Expression ; Genome, Human ; Granulocyte-Macrophage Colony-Stimulating Factor ; Granulocytes ; Humans ; Interleukin-12 ; Interleukin-13 ; Interleukin-3 ; Nutrigenomics ; Obesity ; Oxidoreductases ; Protein Array Analysis ; Tea ; Vascular Endothelial Growth Factor A

OBJECTIVES: To investigate the effect of granulocyte colony stimulating factor (G-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF) on expression of matrix metalloproteinase-2, 9 (MMP-2, 9) mRNA in mouse embryos. Materials and METHOD: From October 1997 to December 1998, morula stage mouse embryos were cultured for 48 hours with G-CSF and GM-CSF at concentrations of 0.1 pg/ml, 1 pg/ml, 10 pg/ml, 100 pg/ml, 1 ng/ml and 10 ng/ml, respectively. Embryos not treated with G-CSF or GM-CSF were served as control. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of MMP-2, 9 mRNA in developed blastocysts. Following reverse transcription, strategically designed nested primers, optimized for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Results were analyzed with Kolmogorov-Smirnov test and analysis of variance (ANOVA). The statistical significance was defined as p< 0.05. RESULTS: The relative quantities (relative volume x intensity) of MMP-2 mRNA expressed in embryos of all G-CSF treatment groups were significantly increased than in the control, especially in 10, 100 pg/ml and 1 ng/ml treatment groups. The relative quantities of MMP-2 mRNA in all GM-CSF treatment groups were also significantly increased than in the control, especially in 100 pg/ml treatment group. The relative quantities of MMP-9 mRNA of all GM-CSF treatment groups except 10 ng/ml group were significantly increased than in the control, especially 10, 100 pg/ml and 1 ng/ml treatment group. However, the relative quantity of MMP-9 mRNA was significantly increased in only 10 ng/ml G-CSF treatment group than in the control and other treatment groups. CONCLUSION: This study suggests that G-CSF and GM-CSF may increase the m-RNA expression of MMP-2 or 9 in mouse blastocysts with the concentration-specific manner.
Animals ; Blastocyst ; Colony-Stimulating Factors* ; Digestion ; DNA, Complementary ; Embryonic Structures* ; Granulocyte Colony-Stimulating Factor ; Granulocyte-Macrophage Colony-Stimulating Factor* ; Granulocytes* ; Matrix Metalloproteinase 2* ; Mice* ; Morula ; Reverse Transcription ; RNA, Messenger ; Sensitivity and Specificity ; Sequence Analysis

BACKGROUND: Transplantation of allogeneic peripheral blood stem cells (PBSCs) may have advantage over bone marrow transplantation with regards to the speed of hematopoietic and immunologic recovery. Recently to overcome the need for multiple leukaphereses to collect enough PBSCs for autologous transplantation, large volume leukaphereses (LVL) are used to process multiple blood volumes per session. Experience with this technique in healthy individuals after mobilization with colony stimulating factor (CSF) is limited. We have investigated the efficacy and safety of LVL for the collection of G-CSF and GM-CSF mobilized PBSCs from healthy donors. METHODS: This study was done on 40 healthy donors who were mobilized with G-CSF and GM-CSF for allogeneic peripheral blood stem cells transplantation (allo-PBSCT). After 5 days of mobilization treatment, PBSCs were collected by LVL with Fenwal CS-3000 Plus (Baxter Co, USA). In LVL, heparin was administered in addition to ACD-A. Patients underwent of LVL procedures daily to obtain a target cell dose of >or= 4x10(8)/kg MNCs and >or= 6x10(6)/kg CD34+ cells. RESULTS: 66 LVL procedures were done on 40 donors. Of these donors, 31 (77.5%) reached the collection target with one leukapheresis. The product per LVL contained a mean 5.79+/-2.47 10(8)MNCs/kg and 11.6+/-10.62x10(6) CD34+ cells/kg respectively. Mean percentages of MNC were 79.88+/-22.15% and collection efficiencies of MNCs were inversely related to the starting MNC count (r=-0.536, P<0.001). After LVL, although none of the donors exhibited bleeding complications, platelets decreased from 187.4+/-52.68x10(3)/microL to 74.88+/-13.7x10(3)/microL and activated partial thromboplastin time (APTT) prolonged from 29.13+/-3.77 seconds to 67.51+/-54.26 seconds. CONCLUSION: We conclude that LVL after mobilization treatment with G-CSF and GM-CSF in normal healthy donors was tolerable and efficient methods for PBSCs collection, but long-term risk of adverse effects in normal donors needs to be carefully addressed by individual institutions as well as national and international registries.
Autografts ; Blood Volume ; Bone Marrow Transplantation ; Colony-Stimulating Factors ; Granulocyte Colony-Stimulating Factor ; Granulocyte-Macrophage Colony-Stimulating Factor ; Hemorrhage ; Heparin ; Humans ; Leukapheresis* ; Partial Thromboplastin Time ; Registries ; Stem Cells* ; Tissue Donors ; Transplantation, Autologous

BACKGROUND: The use of colony stimulating factor (CSF) has increased to mobilize hematopoietic progenitor cells for allo-PBSCT. The most effective mobilization regimen has not yet defined. The authors analyzed the results of the mobilized PBSC collection through large volume leukapheresis from 38 normal healthy donors using three different regimens, namely, a single regimen with GM-CSF (Leucogen ), a concurrent use of GM-CSF and G-CSF (Leucostim ), and a sequential regimen with GM-CSF followed by G-CSF. METHODS: This study was done on 38 healthy donors from Sep. 1998 to Jan. 2001. One donor was mobilized with G-CSF alone, 9 with GM-CSF alone, 20 with concurrent regimen and 8 with sequential regimen. After 5 days of mobilization treatment, PBSCs were collected by large volume leukapheresis through femoral vein catheter. We compared the results of each collected progenitor cells and observed the side effects. RESULTS: The average WBC count before apheresis was 22.6+-11.0x103/uL and circulating CD34+cell percent was 1.31+-2.24%. Total 66 times with an average of 1.46+-0.61 of largevolume leukapheresis were performed for the 37 donors. The mean collected MNC count was 4.61+-2.77x108/kg, CD3+cell count was 2.95+-1.82x108/kg and CD34+cell count was 9.76+-12.42x106/kg. A significant side effect observed after large volume leukapheresis was thrombocytopenia showing decrease from 199.1+-52.2 to 80.7+-25.2x103/uL without any bleeding tendency. The mean collected MNC counts provoed to be significantly higher in combination groups with GM-and G-CSF than GM-CSF alone (P<0.05). The CD34+cell counts showed to be statistically higher in a sequential group compared to the concurrent and single regimen groups (P<0.05). CONCLUSION: A mobilization protocol with combination regimens of GM-CSF and G-CSF seemed to be superior to a single regimen with GM-CSF. Large volume leukapheresis through femoral vein catheter after mobilization with combination regimens of GM-and G-CSF in normal healthy donors was safe and proved to be an excellent. method to harvest stem cells.
Blood Component Removal ; Catheters ; Colony-Stimulating Factors ; Femoral Vein ; Granulocyte Colony-Stimulating Factor* ; Granulocyte-Macrophage Colony-Stimulating Factor* ; Hematopoietic Stem Cells ; Hemorrhage ; Humans ; Leukapheresis* ; Stem Cells* ; Thrombocytopenia ; Tissue Donors*

PURPOSE: Neutrophils are central to the defences against bacterial infection, and in neonates the number of neutrophils are decreased due to inhibited production and phagocytic function. This induces high mortality rates in infants suffering from neonatal sepsis. Exogenous GM-CSF can increase the number of neutrophils and improve the phagocytic function. To establish the most cost effective dose of exogenous granulocyte-macrophage colony-stimulating factor in infected neonates, we divided infected patients into two groups. The serum level of granulocyte-macrophage colony stimulating factor, white blood cell count and absolute granulocyte count were compared. METHODS: This study included 22 infants with infection, admitted to the neonatal intensive care unit of Chungnam National University Hospital, between February 1998 and September 1999. Infected infants were divided into two treatment groups with exogenous GM-CSF 3 microgram/kg/day & 10 microgram/kg/day. The total WBC count, the absolute granulocyte count and the serum GM-CSF concentration of peripheral blood before use of GM-CSF, and those of 2nd, 5th and 7th day after use were compared. RESULTS: In 3 microgram/kg/day group, WBC count and the absolute granulocyte count and the serum GM-CSF concentration reached a peak after the 2nd injection of GM-CSF. In 10 microgram/kg/day group, the WBC count and the absolute granulocyte count increased gradually until 7th day. There was tendency for the total WBC counts, the absolute granulocyte count and the serum GM-CSF concentration of peripheral blood in 10 microgram/kg group to increase more than those of 3 microgram/kg group, after exogenous GM-CSF treatment. CONCLUSION: In neonates, we propose the exogenous GM-CSF treatment 10 microgram/kg/day as being more effective than 3 microgram/kg/day protocol, which is one of the safest and most effective methods to increase the total WBC count, the absolute granulocyte count and the serum GM-CSF concentration of peripheral blood.
Bacterial Infections ; Chungcheongnam-do ; Colony-Stimulating Factors ; Granulocyte-Macrophage Colony-Stimulating Factor* ; Granulocytes ; Humans ; Infant ; Infant, Newborn ; Intensive Care, Neonatal ; Leukocyte Count ; Leukocytes ; Mortality ; Neutrophils ; Sepsis

Idiopathic pulmonary alveolar proteinosis (PAP) is a rare disorder characterized by surfactant component accumulation in the alveolar space. Idiopathic PAP has recently been recognized as a autoimmune disease of impaired alveolar macrophage function caused by autoantibodies against granulocyte-macrophage colony-stimulating factor (GM-CSF). While whole lung lavage has been the standard treatment, not every patient shows a complete response. Subcutaneous injection or inhalation of GM-CSF is another promising treatment option for PAP. A 45-year-old patient visited our hospital for dyspnea, he was diagnosed as PAP and underwent whole lung lavage. Eighteen months later, the patient had not achieved complete remission in despite of initial response. After then he was administered with GM-CSF (5 microgram/kg/day, subcutaneous injection) for fivetimes a week during 2 months. Nine months later, the abnormal shadows in high-resolution computed tomography (HRCT) decreased and the patient fully recovered in forced vital capacity. After 60 months, the HRCT scan showed complete remission of PAP.
Autoantibodies ; Autoimmune Diseases ; Bronchoalveolar Lavage ; Colony-Stimulating Factors ; Dyspnea ; Granulocyte-Macrophage Colony-Stimulating Factor ; Humans ; Inhalation ; Injections, Subcutaneous ; Lung ; Macrophages, Alveolar ; Middle Aged ; Pulmonary Alveolar Proteinosis ; Vital Capacity

OBJECTIVE: Granulocyte-macrophage colony stimulating factors known to be secreted in murine and human reproductive tract. The development of human, bovine and murine embryos could be promoted by addition of GM-CSF in culture medium. However, the pregnancy and implantation rate of embryos cultured in GM-CSF have not been evaluated. The aim of this study was to assess the effect of GM-CSF in embryo development, pregnancy and implantation rate. METHODS: A total of 191 IVF cycles were divided into control and GM-CSF supplement group (control =96, GM-CSF=95). The embryos were cultured for three day with or without 2 ng/ml of recombinant human GM-CSF. The quality of embryo, developmental velocity, pregnancy and implantation rates were compared. RESULTS: There was no difference in age, number of gonadotropin ampules used, number of oocytes and fertilization. The number of ICSI cycle was higher in GM-CSF group. In GM-CSF group, G-1 grade embryos were the highest in proportion (56.4%), while G-2 grade embryos were highest (44.3%) in control group. The developmental velocity of embryos were not different between GM-CSF and control group. The pregnancy and implantation rates were significantly higher in GM-CSF group than control (47.4% vs. 33.3%, 17.0% vs. 11.1% respectively). CONCLUSION: By adding GM-CSF in culture medium, the quality of embryo, pregnancy and implantation rate could be improved.
Colony-Stimulating Factors ; Embryonic Development ; Embryonic Structures ; Female ; Fertilization ; Gonadotropins ; Granulocyte-Macrophage Colony-Stimulating Factor* ; Humans* ; Oocytes ; Pregnancy ; Sperm Injections, Intracytoplasmic