• KSBB Journal
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• v.30 no.2
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• pp.82-90
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• 2015

Plant cell immobilization can protect plant cells from shear forces and increase the stability of gene. An additional advantage of immobilization is the easiness for performing continuous culture with cell recycling. Therefore plant cell immobilization can overcome the limitations of plant cell applications. In addition, target protein should be selected from pharmaceutical proteins to get rid of low expression level problem. The enhanced production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was investigated in immobilized Nicotiana tabacum suspension cell cultures. When the cells were immobilized in polyurethane foam, specific production of hGM-CSF was higher than that in alginate bead immobilization. Optimum continuous culture condition was the addition of 60 g/L sucrose in growth media with exchanging media every 6 day. Under the same condition, specific hGM-CSF production was 7 times higher in a 500-mL spinner flask than that in 100-mL Erlenmeyer flasks. Therefore, development of an effective immobilization process would be possible when the advantage of easy cell recycling was used. Consequently, enhanced production of target proteins could be possible in immobilized continuous cultures when the advantages of immobilization were applied.

• Journal of Korean Medicine Rehabilitation
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• v.15 no.1
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• pp.25-37
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• 2005

목적 : 진피(Citus Unshiu, CU)는 실험적으로 항산화, 항알러지, 항종양 효과 등이 있다고 보고되고 있다. 본 연구에서는 비만세포에서 PMA와 A23187에 의하여 유도된 싸이토카인의 증가에 대한 진피 추출물의 억제 효과와 그 기전을 알아보고자 한다. 방법 : PMA+A23187에 의한 싸이토카인 생성에 대한 진피 추출물의 억제효과 기전을 조사하기 위하여 진피를 처리한 후 $IL-1{\beta}$, IL-8, $TNF-{\alpha}$의 생산 및 혈관내피성장인자 (VEGF), 과립구 대식구 군집 자극 인자 (GM-CSF), 저산소 유도 인자(HIF-1)의 발현을 조사하였다. 결과 : 실험에서 PMA와 A23187을 처리한 경우 대조군에 비하여 $IL-1{\beta}$, IL-8, $TNF-{\alpha}$의 생산을 유의하게 증가시켰으나 진피 추출물을 처리한 군에서는 유의하게 억제되었다. 특히 $IL-1{\beta}$의 생성은 $103.7{\pm}5%$, IL-8 생성은 $34.6{\pm}7%$, $TNF-{\alpha}$의 생성은 $85.9{\pm}4.5%$ 억제되었다. (P<0.05). 또한 진피 추출물은 PMA와 A23187에 의하여 증가한 VEGF, GM-CSF, $HIF-1{\alpha}$의 발현 증가를 억제하였다 (약 90.9%의 VEGF, 61.6%의 GM-CSF). 결론 : 이러한 결과는 진피가 염증반응에 대한 HIF-1의 억제자로 작용할 수 있으면, 진피가 비만세포 매개성 염증 질환 치료제의 후보 물질이 될 것으로 사료된다.

• IMMUNE NETWORK
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• v.5 no.2
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• pp.105-109
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• 2005

Background: Dendritic cells (DCs) are the most potent APCs (antigen-presenting cells) and playa critical role in immune responses. Galectin-3 is a biological lectin with a beta-galactoside binding affinity. Recently, proteomic analysis revealed the presence of galectin-3 in the exosome of mature DCs. However, the expression and function of galectin-3 in DCs remains unclear yet. Methods: We used bone marrow-derived DCs of mouse and showed the expression of galectin-3 in DCs by using flow cytometry analysis and Western blot analysis. Results: Galectin-3 was determined as single band of 35 kDa in Western blot analysis. Flow cytometry analysis showed the major growth factor for DCs, granulocyte-macrophage colony stimulating factor (GM-CSF) and maturing agents, anti-CD40 monoclonal antibody (mAb) and lipopolysaccharide (LPS) consistently increased the intracellular expression of galectin-3 in DCs compared to medium alone. In addition, DCs treated with maturing agents did marginally express galectin-3 on their surface. Conclusion: This study suggests that galectin-3 in DCs may be regulated by critical factors for DC function.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.95-95
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• 1993

대상환자는 15예로, 14예에서 평가가능하였다. 남녀비는 8:6, 중앙연령 32세(10-70세)이었고, 대상질환은 악성골육종 7예, 악성임파종 2예, 위암2예, 폐암 2예, 그리고 자궁악성육종 1예였다. CSF 50 ug/$m^2$ 3예, 100 ug/$m^2$ 3예,150 ug/$m^2$ 3예, 250 ug/$m^2$ 3예,350 ug/$m^2$ 3예, 500 ug/$m^2$ 6예, 700 ug/$m^2$ 용량 3예에서 시행되었다. 1주기 시행한 환자는 7예, 2주기 5예, 3주기, 4주기 각각 1예씩이었다. 부작용은 50-150ug/$m^2$에서 WHO grade I의 발열, 전신쇠약, 식욕부진 등이 관찰되었으나 grade II이상의 부작용은 없었다. 250 ug/$m^2$ 이상의 용량에서도 grade II의 발열이 관찰되었을 뿐 다른 중증의 부작용은 관찰되지 않았다. 최고용량인 700 ug/$m^2$ 에서도 grade II의 발열외의 중한 부작용은 관찰되지 않았다. 각 용량에 따른 백혈구 증가율(%투여제2일/투여제1일)은 130-500% 이었다. 시행된 약동태는 CSF 투여 2-4 시간 후 최고치 (0.42-15.4 ng/ml)를 나타내고 투여 12시간까지 0.2-2 ng/ml의 농도로 지속되었다. 소변내 CSF 배설량은 총투여량의 1% 미만이었다. CSF중화항체는 전예에서 검출되지 않았다.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.68-68
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• 1993

대상환자는 15예로, 14예에서 평가 가능하였다. 남녀비는 8:6, 중앙연령 32세(10-70세)이었고, 대상질환은 악성골육종 7예, 악성임파종 2예, 위암2예, 폐암 2예, 그리고 자궁악성육종 1예였다. CSF 50 ug/$m^2$ 3예, 100 ug/$m^2$ 3예,150 ug/$m^2$ 3예, 250 ug/$m^2$ 3예, 350 ug/$m^2$ 3예, 500 ug/$m^2$ 6예, 700 ug/$m^2$ 용량 3예에서 시행되었다. 1주기 시행한 환자는 7예, 2주기 5예, 3주기, 4주기 각각 1예씩이었다. 부작용은 50-150ug/$m^2$에서 WHO grade I의 발열, 전신쇠약, 식욕부진등이 관찰되었으나 grade II이상의 부작용은 없었다. 250 ug/$m^2$ 이상의 용량에서도 grade II의 발열이 관찰되었을 뿐 다른 중증의 부작용은 관찰되지 않았다. 최고용량인 700 ug/$m^2$ 에서도 grade II의 발열 외의 중한 부작용은 관찰되지 않았다. 각 용량에 따른 백혈구 증가율(%투여제2일/투여제1일)은 130-500% 이었다. 시행된 약동태는 CSF 투여 2-4 시간후 최고치 (0,42-15.4 ng/ml)를 나타내고 투여 12시간까지 0.2-2 ng/ml의 농도로 지속되었다. 소변내 CSF 배설량은 총투여량의 1% 미만이었다. CSF 중화항체는 전예에서 검출되지 않았다.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.320-323
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• 2003

The effects of pH on the binding of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) expressed from transgenic plant cell suspensions to cationic and anionic exchange resins were investigated. In terms of stability, the optimum pH was found to be 5-7. In the case of using buffer exchange, when CM-sepharose was used as a cationic exchange resin, the best binding pH was 4.8 (77%) and when DEAE-sepharose was used as an anionic exchange resin, the best binding pH was 5.5 (74%). Without using buffer exchange, the optimum pH was 4.6 and the adsorption yield was 84%. From these results, a possibility of overcoming the degradation and instability of secreted protein product by in firm adsorption was found.

• The Korean Journal of Physiology and Pharmacology
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• v.16 no.4
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• pp.243-247
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• 2012

Recent studies suggest that immunization with autologous dendritic cells (DCs) results in protective immunity and rejection of established tumors in various human malignancies. The purpose of this study is to determine whether DCs are generated from peripheral blood mononuclear cells (PBMNs) by using cytokines such as F1t-3 ligand (FL), granulocyte macrophage-colony stimulating factor (GM-CSF), IL-4, and TNF-${\alpha}$, and whether cytotoxic T cells activated against the thyroid cancer tissues by the DCs. Peripheral blood was obtained from 2 patients with thyroid cancer. DCs were established from PBMNs by culturing in the presence of FL, GM-CSF, IL-4, and TNF-${\alpha}$ for 14 days. At day 14, the differentiated DCs was analyzed morphologically. The immunophenotypic features of DCs such as CDla, CD83, and CD86 were analyzed by immunofluorelescence microscopy. At day 18, DCs and T cells were incubated with thyroid cancer tissues or normal thyroid tissues for additional 4 days, respectively. DCs generated from the PBMNs showed the typical morphology of DCs. Activated cytotoxic T lymphocytes (CTLs) were observed also. DCs and the CTLs were attached to the cancer tissues on scanning electron microscope. The DCs activated the CTLs, which able to specifically attack the thyroid cancer. This study provides morphologic evidence that the coculture of T cells/cancer tissues activated the T cells and differentiated CTLs. The CTLs tightly adhered to cancer tissues and lysed cancer tissues vigorously. Therefore DCs could be used as potential vaccines in the immunotherapy.

• Food Science and Biotechnology
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• v.14 no.1
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• pp.147-151
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• 2005

Purified compound with intestinal immune system-modulating properties, GWE-2c, was isolated from methanol extract of Gelidium amansii by sequential procedures with silica gel column, LH-20 Sephadex gel column, and thin-layer chromatographies. In the presence of GWE-2c, strong immunoactivity in Peyers patch cell-mediated bone marrow cells was observed in vitro. In vivo intestinal immune-modulating activity was also enhanced by crude phenolic compound (GWE) of G. amansii in a dose-dependent manner. Investigation of production of several cytokines in Peyer's patch cells upon stimulation with GWE in vivo revealed the levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-6 increased. Results suggest that the phenolic compound from G. amansii represents immunopotentiator and biological response modifier at in vitro and in vivo levels.

• Tuberculosis and Respiratory Diseases
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• v.80 no.1
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• pp.77-82
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• 2017

Background: Delayed hypersensitivity plays a large role in the pathogenesis of tuberculous pleural effusion (TPE). Macrophages infected with live Mycobacterium tuberculosis (MTB) increase the levels of adenosine deaminase2 (ADA2) in the pleural fluid of TPE patients. However, it is as yet unclear whether ADA2 can be produced by macrophages when challenged with MTB antigens alone. This study therefore evaluated the levels of ADA2 mRNA expression, using monocyte-derived macrophages (MDMs) stimulated with MTB antigens. Methods: Purified monocytes from the peripheral blood mononuclear cells of healthy volunteers were differentiated into macrophages using granulocyte-macrophage colony-stimulating factor (GM-CSF) or macrophage colony-stimulating factor (M-CSF). The MDMs were stimulated with early secretory antigenic target protein 6 (ESAT6) and culture filtrate protein 10 (CFP10). The mRNA expression levels for the cat eye syndrome chromosome region, candidate 1 (CECR1) gene encoding ADA2 were then measured. Results: CECR1 mRNA expression levels were significantly higher in MDMs stimulated with ESAT6 and CFP10, than in the unstimulated MDMs. When stimulated with ESAT6, M-CSF-treated MDMs showed more pronounced CECR1 mRNA expression than GM-CSF-treated MDMs. Interferon-${\gamma}$ decreased the ESAT6- and CFP10-induced CECR1 mRNA expression in MDMs. CECR1 mRNA expression levels were positively correlated with mRNA expression of tumor necrosis factor ${\alpha}$ and interleukin 10, respectively. Conclusion: ADA2 mRNA expression increased when MDMs were stimulated with MTB antigens alone. This partly indicates that pleural fluid ADA levels could increase in patients with culture-negative TPE. Our results may be helpful in improving the understanding of TPE pathogenesis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.6
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• pp.738-744
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• 2013

The purpose of this study is to investigate effects of White Ginseng-Ejung-tang water extract (EJ) on production of various cytokines such as interleukin (IL)-2, IL-5, IL-6, IL-10, IL-12p70, macrophage inflammatory protein (MIP)-2, vascular endothelial growth factor (VEGF), keratinocyte-derived chemokine(KC), tumor necrosis factor (TNF)-${\alpha}$, and granulocyte macrophage colony-stimulating factor (GM-CSF) in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). Levels of cytokines were measured by High-throughput multiplex bead array cytokine assay based on xMAP (multi-analyte profiling beads) technology. EJ significantly decreased levels of IL-2, IL-12p70, IL-5, MIP-2 for 24 h incubation at the concentrations of 25, 50, and 100 ${\mu}g/mL$ in LPS-induced RAW 264.7 (P < 0.05). EJ significantly decreased levels of IL-6 at the concentrations of 50 and 100 ${\mu}g/mL$ (P < 0.05). EJ significantly decreased levels of IL-10 and VEGF at the concentrations of 25 and 100 ${\mu}g/mL$ (P < 0.05). EJ significantly decreased levels of KC at the concentrations of 100 ${\mu}g/mL$ (P < 0.05). EJ did not show any significant effect on TNF-${\alpha}$ and GM-CSF production. These results suggest that EJ has anti-inflammtory property related with its inhibition of IL-2, IL-5, IL-6, IL-10, IL-12p70, MIP-2, VEGF, and KC production in LPS-induced macrophages.