• IMMUNE NETWORK
• /
• 제6권4호
• /
• pp.169-178
• /
• 2006

Background: Adipose tissues were initially introduced as energy storages, but recently they have become famous as an endocrine organ which produces and secretes various kinds of molecules to make physiologic and metabolic changes in human body. It has been studied that these molecules are secreted in abundance as the adipose tissue becomes bigger along with obesity. Furthermore, it has been found that they are mediating systemic inflammation and generation of metabolic diseases such as type 2 diabetes and atherosclerosis. On the basis of these, we studied previous papers which have been researched about the interaction between preadipocytes and macrophages, adipose tissues and lymph nodes, and adipose tissue secreting molecules. Results: Firstly, preadipocytes and macrophages are expressing similar transcriptomes and proteins, and preadipocytes can be converted to mature macrophages which have phagocytic activity. Moreover, the monocytes, which initially located in the bone marrow, are filtrated to the adipose tissue by monocyte chemotatic protein-1 and are matured to macrophages by colony stimulating factor-1. Secondly, adipose tissues and their associated lymph nodes are interacting each other in terms of energy efficiency. Lymph nodes promote lipolysis in adipose tissues, and polyunsaturated fatty acids in adipocytes become energy sources for dendritic cells. Lastly, adipose tissues produce and secrete proinflammatory molecules such as leptin, adiponectin, TNF-${\alpha}$, IL-6, and acute phase proteins, which induce the inflammation and potentially generate metabolic diseases. Conclusion: According to these, we can link adipose tissues to inflammation, but we need to affirm the actual levels and roles of adipose tissue-derived proinflammatory molecules in human body.

• KSBB Journal
• /
• 제30권2호
• /
• pp.82-90
• /
• 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.

• IMMUNE NETWORK
• /
• 제3권2호
• /
• pp.96-102
• /
• 2003

Background: Granulocyte-macrophage colony-stimulating factor (GM-CSF) gene-transduced tumor cell vaccines induce very potent systemic anti-tumor immunity in preclinical and clinical models. Our previous phase I clinical trial in patients with metastatic renal cell carcinoma (RCC) has demonstrated both immune cell infiltration at vaccine sites and T cell-mediated delayed-type hypersensitivity (DTH) response to whole tumor cell vaccines. Methods: To investigate the immune responses to autologous genetically- modified tumor cell vaccines, tumor-specific $CD8^+$ T cell lines were generated from peripheral blood lymphocytes (PBL) of a RCC patient 1.24 by repeated in vitro stimulation with either B7.1-transduced autologous RCC tumor cells or B7.1-transduced autologous tumor cells treated with interferon gamma ($IFN{\gamma}$), and cloned by limiting dilution. Results: Among several RCC-specific cytotoxic T lymphocytes (CTLs), a $CD4^+/CD8^+$ double positive T cell clone (17/A2) appeared to recognize $IFN{\gamma}$-treated autologous RCC restricted by HLA-B39. The 17/A2 also recognized other HLA-B39 positive RCC tumor cells after $IFN{\gamma}$ treatment. Conclusion: These results demonstrate that autologous RCC vaccination successfully generates the tumor-specific CTL 17/A2, and suggest that the presentation and recognition of the tumor antigen by the 17/A2 might be upregulated by $IFN{\gamma}$.

• KSBB Journal
• /
• 제16권6호
• /
• pp.568-572
• /
• 2001

빛은 식물에서 성장과 발달을 비롯한 다양한 생리화학적인 역할은 지닌다. 본 연구는 hGM-CSF 유전자가 도입된 형질전한 담배의 callus를 현탁배양하여 hGM-CSF를 생산할 때에 빛을 조사하는 시간에 따른 hGM-CSF의 생산에 미치는 영향을 확인하고자 실시하였다. 24시간 명배양, 18시간 명배양과 암배양을 실시하여 세포성장과 분비된 총단백질, hGM-CSF 생산량을 비교 관찰하였다. 세포의 성장은 24시간 명배양일 때 건조중량이 14.4 g/L로 가장 높았다. 분비된 총단백질의 양은 세가지 경우에서 큰 차이를 관찰할 수가 없었지만, 단위 세포당 분비된 총단백질의 양은 암배양이 다른 것에 비해 1.5배 가량 높았다. hGM-CSF의 생산은 18시간 명배양 조건이 가장 좋았으며 최대생산량이 495.5ug/L에 이르렀다. 또한 분비된 총단백질에서 hGM-CSF가 차지하는 비율은 18시간 명배양이 24시간 명배양에 비해 최대 1.8배 높았다.

• KSBB Journal
• /
• 제30권3호
• /
• pp.103-108
• /
• 2015

Production of foreign proteins by transgenic plant cell cultures has several advantages such as post-translational modification, low risk of product contamination and low-cost production and purification. However, target proteins are degraded by extracellular proteases existing in the media. A solution to this problem is the use of perfusion culture and ion exchange chromatography for the application of integrated bioprocess using in situ recovery. With this method, production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was investigated in this study. First, optimization of cell concentration during the induction phase for the production of hGM-CSF was examined. As cell concentration increased, the level of hGM-CSF was decreased due to the presence of extracellular proteases. Induction using sugarfree media produced 33% more hGM-CSF. The effects of pH on the binding of hGM-CSF to cationic and anionic exchange resins were also investigated. In terms of stability, optimal pH was found to be 5~7. In the case of using buffer exchange when CM-Sepharose was used as a cationic exchange resin, optimal pH for binding was 4.8 and adsorption yield was 77%. When DEAE-Sepharose was used as an anionic exchange resin, it was 5.5 (74%). Without buffer exchange, optimal pH was 4.6 (84%). From these results, an integrated bioprocess using in situ recovery with simultaneous production and separation of foreign protein in transgenic plant cell suspension cultures was found to be feasible.

• Tuberculosis and Respiratory Diseases
• /
• 제81권4호
• /
• pp.305-310
• /
• 2018

Background: Mannose-binding lectin (MBL) deficiency leads to increased susceptibility to infection. We investigated whether serial changes in MBL levels are associated with the prognosis of patients diagnosed with septic shock, and correlated with cytokine levels. Methods: We enrolled 131 patients with septic shock in the study. We analyzed the serum samples for MBL and cytokine levels at baseline and 7 days later. Samples on day 7 were available in 73 patients. Results: We divided the patients with septic shock into four groups according to serum MBL levels (< $1.3{\mu}g/mL$ or ${\geq}1.3{\mu}g/mL$) on days 1 and 7. Patients with low MBL levels on day 1 and high MBL levels on day 7 showed a favorable prognosis for 28-day survival (odds ratio, 1.96, 95% confidence interval, 1.10-2.87; p=0.087). The high MBL group on day 7 showed a significant decrease in monocyte chemoattractant protein 1, interleukin (IL)-$1{\beta}$, IL-6, IL-8, interferon-${\gamma}$, and granulocyte macrophage colony-stimulating factor levels compared with the low MBL group on day 7. Conclusion: The increase in MBL levels of patients with septic shock may suggest a favorable prognosis and attenuate pro-inflammatory and anti-inflammatory responses.

• 대한바이러스학회지
• /
• 제30권2호
• /
• pp.141-150
• /
• 2000

The effectiveness of noninfectious recombinant adenovirus encoding murine granulocyte-macrophage colony stimulating factor (mGM-CSF) for the treatment of Meth A fibrosarcoma was investigated in syngeneic BALB/C model. Meth A and HeLa cells transduced with the recombinant adenovirus (Ad.mGM-CSF) produced substantial amounts of mGM-CSF, while WEH1164 cells transduced with the virus did not produce mGM-CSF. Mice inoculated subcutaneously with $1{\times}10^6$ Meth A cells, followed by injection of Ad.dE1 as a control, developed large tumors that reached a mean tumor size of 22 mm by day 30. However, tumor development and tumorigenicity were significantly inhibited in mice with a single intratumoral injection of Ad.mGM-CSF at $1{\times}10^8\;pfu$. Histological examination of the tumors injected with Ad.mGM-CSF revealed dense infiltrates of neutrophils, histiocytes, lymphocytes, and eosinophils associated with apoptotic cell death. The results suggest that the recombinant adenovirus encoding GM-CSF have a potential use for cancer gene therapy.

• Journal of Plant Biotechnology
• /
• 제30권1호
• /
• pp.97-102
• /
• 2003

hGM-CSF가 식물세포 현탁 배양을 통하여 생산이 가능한지를 조사하기 위하여 hGM-CSF를 포함하고 있는 A. tumerfaciens LBA4404를 가지고 상추에 형질전환시켰다. 형질전환된 상추로부터 캘러스를 유도하여 캘러스를 이용한 세포배양체계를 확립하였다. PCR과 Southern blot analysis 결과 상추에 hGM-CSF 유전자가 도입된 것을 확인하였으며, Northern blot analysis 결과 상추식물체에 hGM-CSF 유전자가 발현됨을 확인하였다. 현탁 배양 세포로부터 분비된 hGM-CSF를 ELISA를 이용하여 측정한 결과 149.0 $\mu\textrm{g}$/L가 생산됨 을 확인하였다 이러한 결과는 상추의 현탁 배양 세포가 hGM-CSF와 같은 치료용 단백질의 생산 숙주로 이용될 수 있음을 보여주었다.

• Molecules and Cells
• /
• 제37권8호
• /
• pp.598-604
• /
• 2014

Fatty acids, important components of a normal diet, have been reported to play a role in bone metabolism. Osteoclasts are bone-resorbing cells that are responsible for many bone-destructive diseases such as osteoporosis. In this study, we investigated the impact of a medium-chain fatty acid, capric acid, on the osteoclast differentiation, function, and survival induced by receptor activator of NF-${\kappa}B$ ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). Capric acid inhibited RANKL-mediated osteoclastogenesis in bone marrow-derived macrophages and suppressed RANKL-induced $I{\kappa}B{\alpha}$ phosphorylation, p65 nuclear translocation, and NF-${\kappa}B$ transcriptional activity. Capric acid further blocked the RANKL-stimulated activation of ERK without affecting JNK or p38. The induction of NFATc1 in response to RANKL was also attenuated by capric acid. In addition, capric acid abrogated M-CSF and RANKL-mediated cytoskeleton reorganization, which is crucial for the efficient bone resorption of osteoclasts. Capric acid also increased apoptosis in mature osteoclasts through the induction of Bim expression and the suppression of ERK activation by M-CSF. Together, our results reveal that capric acid has inhibitory effects on osteoclast development. We therefore suggest that capric acid may have potential therapeutic implications for the treatment of bone resorption-associated disorders.

• 한국식품영양학회지
• /
• 제27권3호
• /
• pp.400-405
• /
• 2014

최근 알로에 베라 섭취에 의한 독성 간염이 보고되고 있으나, 아직까지 알로에 베라의 간에 대한 염증 효과가 명확히 밝혀지지 않았다. 본 연구는 알로에 베라 에탄올 추출물이 간세포의 염증 발현에 미치는 영향과 그 기전을 밝히기 위해 수행되었다. $0.001{\sim}100{\mu}g/mL$의 알로에 베라 추출물을 HepG2 세포에 처리하여 MTT assay를 시행한 결과, 모든 농도에서 세포사멸이 유도되지 않았다. 그러나 모든 농도에서 알로에 베라 추출물은 염증성 사이토카인인 IL-8의 분비를 15.7~25.8%까지 유의적으로 증가시켰다(p<0.05). 또 다른 사이토카인인 M-CSF도 알로에 베라 추출물에 의해 36.3~61.5%까지 유의적으로 분비가 증가되었다(p<0.05). 본 연구 결과는 알로에 베라 추출물이 IL-8과 M-CSF와 같은 염증성 사이토카인 분비기전에 의해 간에 염증을 유발할 수 있음을 보여준다. 또한 알로에 추출물에 의해 유발될 수 있는 간염기전을 제시함으로써, 향후 수행될 추가 실험을 위한 기초 자료로 그 가치가 높을 것으로 사료된다.