• BMB Reports
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• 제46권4호
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• pp.213-218
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• 2013

Members of the colony stimulating factor cytokine family play important roles in macrophage activation and recruitment to inflammatory lesions. Among them, granulocyte-macrophage colony stimulating factor (GM-CSF) is known to be associated with immune response to mycobacterial infection. However, the mechanism through which Mycobacterium tuberculosis (MTB) affects the expression of GM-CSF is poorly understood. Using PMA-differentiated THP-1 cells, we found that MTB infection increased GM-CSF mRNA expression in a dose-dependent manner. Induction of GM-CSF mRNA expression peaked 6 h after infection, declining gradually thereafter and returning to its basal levels at 72 h. Secretion of GM-CSF protein was also elevated by MTB infection. The increase in mRNA expression and protein secretion of GM-CSF caused by MTB was inhibited in cells treated with inhibitors of p38 MAPK, mitogen-activated protein kinase kinase (MEK-1), and PI3-K. These results suggest that up-regulation of GM-CSF by MTB is mediated via the PI3-K/MEK1/p38 MAPK-associated signaling pathway.

• Radiation Oncology Journal
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• 제13권1호
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• pp.79-85
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• 1995

Purpose : To assess the efficacy of recombinant human granulocyte-macrophage colony-stimulating factor(GM-CSF) in the neutropenia by radiotherapy. Materials and Methods : Eleven patients with various solid tumor were treated with a daily subcutaneous dose of GM-CSF(3-7microgram/kg) for 5days during the radiotherapy. Before and during the course of the study all the patients were monitored by the recording of physical examination, the complete blood count with differential and reticulocyte count and liver function test. Eight patients received prior or concurrent chemotherapy. Results : In 10 patients, the neutrophilic nadir was significantly elevated and the lenght of time that Patients had a neutrophil count below $10^3/mm^3$ a threshold known to be critical to acquiring infective complications was shortened following GM-CSF injection. A significant rise (two fold or greater) of neutrophil count was seen in 10 of 11 patients. In most patients, discontinuation of GM-CSF resulted in a prompt return of granulocyte counts toward baseline. However the neutrophil count remained elevated over $10^3/mm^3$ during radiation therapy, and radiotherapy delays were avoided. Other peripheral blood components including monocytes and platelets also increased after GM-CSF treatment. No significant toxicity was encountered with subcutaneous GM-CSF treatment. Conclusion : GM-CSF was well tolerated by subcutaneous route and induced improvement in the neutropenia caused by radiotherapy.

• Biomolecules & Therapeutics
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• 제5권3호
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• pp.316-322
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• 1997

CJ-50001 is a recombinant granulocyte-colony stimulating factor (rG-CSF) synthesized by recombi-nant DNA technology using E. coli as an expression system. The general pharmacological properties of CJ-50001 were evaluated in mice, rats, dogs and isolated guinea pig ileum. The doses are 100, 300 and 1, 0007g/kg, i.v. for mice and rats, 1, 10 and 100$\mu$g/kg, 1.v. for dogs and 1 and 10$\mu$g/ml for isolated guinea pig ileum. Intravenous administration of CJ-50001 at this dose range did not affect general behavior, central nervous system, smooth muscles, gastrointestinal system, cardiovascular and respiratory system and water and electro-lytes excretion. In summary, CJ-50001 had no harmful pharmacological erect in these studies even up to the 200-fold expected clinical dose, 2507g/man.

• Biomolecules & Therapeutics
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• 제2권3호
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• pp.281-285
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• 1994

Neutropenia is a major dose-limiting factor in cancer chemotherapy diminishing its usefulness and increase patient's susceptibility to infectious disease. Some recombinant human granulocyte colony stimulating factors(rhG-CSFs) are in use to reduce the risk of this serious side effect. In this study, we examined the pharmacological properties of DA-3030, a rhG-CSF expressed in E. coli. DA-3030 100 and $\mu\textrm{g}$/kg, i. v., had no significant effect on the central nervous, gastrointestinal system in mice and cardiovascular system in rabbits, but it slightly inhibited the spontaneous motility of isolated nonpregnant uterus in rats. It also had no influence on excretion of urinary electrolytes. DA-3030 administered for successive 3 days increased the blood WBC count in zymosan air pouch inflammed rats and in normal rats. These results indicate that DA-3030 has little side effects in animals.

• Biomolecules & Therapeutics
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• 제2권3호
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• pp.256-259
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• 1994

This study was performed to evaluate the acute toxicity of DA-3030(granulocyte-colony stimulating factor, G-CSF) in mice and rats via intragastrical and intravenous routes. DA-3030(G-CSF) in the acute toxicity study did not induce any toxic signs in the mice and rats in mortalities, clinical findings, body weights and gross findings. It is suggested that LD$_{50}$ values in mice and rats would be >13, 800 $\mu\textrm{g}$/kg in the oral route and >6, 900 $\mu\textrm{g}$/kg in the intravenous route.e.

• Archives of Pharmacal Research
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• 제23권3호
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• pp.252-256
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• 2000

A thymic stromal cell line, TFGD, was established from a thymic tumor mass developed spontaneously in p53 knock out mouse, and was found to produce cytokines that could induce bone marrow hematopoietic stem cells (HSCs) to differentiate into macrophages. The cytokines produced by the TFGD line were assessed by immunoassays. High level of macrophage-colony stimulating factor (M-CSF) and interleukin (IL)-6 was detected in the TFGD-culture supernatant, whereas granulocyte/macrophage-colony stimulating factor (GM-CSF), IL-3, IL-4, IL-5, IL-13, or interferon (IFN)-$\gamma$ was undetectable. Blocking experiments showed that anti-M-CSF monoclonal antibody could neutralize the differentiation-inducing activity shown by the TFGD-culture supernatant. Dot blot analysis of the total RNA isolated from the cultured fetal thymic stromal cells showed that M-CSF transcripts were expressed in the normal thymus. These observations, together with the earlier finding that M-CSF plus IL-6 is the optimal combination of cytokines for the induction of macrophage differentiation from HSCs in vitro, may indicate that thymic macrophages could be generated within the thymus by cytokines involving M-CSF.

• Clinical and Experimental Reproductive Medicine
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• 제30권1호
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• pp.5-14
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• 2003

Objective: Present study was aimed to verify the effect of granulocyte macrophage-colony stimulating factor (GM-CSF) in the preimplantation development of mouse embryos and the involvement of the mitogen activated protein kiase (MAPK) in the GM-CSF signaling. Methods: Two-cell embryos were cultured for 96 h in the presence or absence of GM-CSF (0, 0.4, 2, 10 ng/ml) and PD98059, a MEK inhibitor (10 ${\mu}M$). Morphological development, cell number per blastocyst, and apoptotic nuclei, were eamined. MAPK activity of embryonic immunoprecipitate by MAPK (ERK1/2) antibody was measured by in vitro phosphorylation of myelin basic protein. Results: At post hCG 122 h the embryonic development among the experimental groups was significantly different (p=0.018). The rate of blastocyst development and cell number per embryo were the highest in 2 ng/ml GM-CSF treatment group. The percent of apoptotic cells of the GM-CSF-treated embryos was the lowest among the group. In blastocysts, GM-CSF treatment transiently increased MAPK activity. PD098059 attenuated the effect of GM-CSF on the morphological development, increase in cell number per blastocyst, down regulation of apoptosis, and upregulation of MAPK activity, suggesting that activation of MAPK cascade possibly mediated the embryotropic effect of GM-CSF. Conclusion: This result suggested that GM-CSF potentiated the development of preimplantation mouse embryos by activation of MAPK.

• Tuberculosis and Respiratory Diseases
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• 제67권6호
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• pp.569-573
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• 2009

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 ${mu}g/kg/day$, 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.

• Biomolecules & Therapeutics
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• 제2권3호
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• pp.292-297
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• 1994

This study was conducted to investigate antigenic potential of DA-3030, a recombinant human granulocyte-colony stimulating factor, in guinea pigs and mice. In the active systemic anaphylaxis test, the guinea pigs sensitized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone did not show any anaphylactic reaction. In the homologous passive cutaneous anaphylaxis reaction, anti-DA-3030 antibody was not detected in guinea pigs sensitized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone. On the other hand, the guinea pigs sensitized with 12.5 $\mu\textrm{g}$/heed of DA-3030 incorporated in Freund's complete adjuvant(FCA) or 1 mg/head of ovalbumin incorporated in FCA showed anaphylactic reaction. Anti-DA-3030 antibody was also detected in those guinea pigs. In immunodiffusion test using the sera sensitized with DA-3030 incorporated in FCA, precipitating antibodies were detected only in the sera sensitized with DA-3030 or DA-3030 incorporated in FCA showed. In 24-hour heterologous PCA reaction with sera of C57BL/6 mice immunized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone, none of the sera showed positive reaction. But sera of the animals immunized with 12.5 $\mu\textrm{g}$/head of DA-3030 incorporated in aluminum hydroxide gel(Alum) or 5 $\mu\textrm{g}$/head of ovalbumin incorporated in alum showed positive PCA reaction. DA-3030 did not cause anaphylactic shock or passive cutaneous anaphylaxis in guinea pigs and mice when given alone although DA-3030 incorporated in FCA or Alum induced anaphylactic shock and passive cutaneous anaphylaxis. From these results, it may be concluded the DA-3030 does not induce systemic allergic reaction when administered alone in its clinical use.

• Journal of Microbiology and Biotechnology
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• 제15권6호
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• pp.1267-1272
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• 2005

The effects of coexpression of GroEL/ES and DnaK/DnaJ/GrpE chaperones on the productivity of the soluble form of human granulocyte colony stimulating factor (hG-CSF) in E. coli were examined. Recombinant hG-CSF protein was coexpressed with DnaK/DnaJ/GrpE or GroEL/ES chaperones under the control of the araB or Pzt-1 promoter, respectively. The optimal concentration of L-arabinose for the expression of DnaK/DnaJ/GrpE was found to be 1 mg/ml. When L-arabinose was added at $OD_{600}$=0.2 (early-exponential phase), soluble hG-CSF production was greatly increased. In addition, it was observed that the DnaK/DnaJ/GrpE and GroEL/ES chaperones had no synergistic effects on preventing aggregation of hG-CSF protein. Consequently, by coexpression of the DnaK/DnaJ/GrpE chaperone, the signal intensity of the hG-CSF protein band in the soluble fraction of cell lysate was increased from $3.5\%\;to\;13.9\%$, and Western blot analysis also revealed about a 4-5-fold increase of production of soluble hG-CSF over the non-induction case of DnaK/DnaJ/GrpE.