급성 골수성 백혈병 환자의 말초혈액 단핵구로부터 수지상세포로의 분화 유도

손상희,이대희,박재선,어완규 고신대학교(의대) 고신대학교 의과대학 학술지 2002 고신대학교 의과대학 학술지 Vol.17 No.1

Background : Dendritic Cells (DCs) are the most potent naturally occuring antigen presenting cells and play an important role in T-cell activation. DCs may be suited for in vivo immunotherapy for its capability to stimulate naive T cell. Effective presentation of tumor antigens is fundamental to strategies aimed at enrolling the immune system in eradication of residual disease after conventional treatments. Myeloid malignancies provide a unique opportunity to derive dendritic cells (DCs), functioning antigen presenting cells, from the malignant cells themselves. These may then co-express leukemic antigens together with appropriate secondary signals and be used to generate a specific, antileukemic immune response. Methods : In this study, peripheral blood mononuclear cells (PBMCs) from 5 patients with acute myeloid leukemia (AML) were cultured with combinations of Flt-3 Ligand (FL), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4), and tumor necrosis factor-a (TNF-a), and development to DCs was assessed. Peripheral blood mononuclear cells (PBMCs) were obtained from 40-60 ml of peripheral blood of patients with acute myeloid leukemia (AML) by Ficolling. Cells were resuspended in X VIVO-20 medium supplemeted with FL (100ng/ml), GM-CSF (100ng/ml), IL-4 (50ng/ml), and TNF-a (20ng/ml) and seeded into T75 culture flasks at 3x10^7/50ml. Results : After 12 days in culture, cells from 5 samples exhibited morphological and immunophenotypic features of DCs, including expression of CD1a, CD83, and CD86. Conclusion : This study indicates that cells with enhanced antigen-presenting ability can be generated from PBMCs of patients with acute myeloid leukemia, that these cells can effectively prime autologous cytotoxic T cells in vitro, and that they may be used as potential vaccines in the immunotherapy of AML.

Zinc Enhances Neutrophil Extracellular Trap Formation of Porcine Peripheral Blood Polymorphonuclear Cells through Tumor Necrosis Factor-Alpha from Peripheral Blood Mononuclear Cells

허주행,김학현,강병택,양만표 한국임상수의학회 2020 한국임상수의학회지 Vol.37 No.5

Neutrophil extracellular trap (NET) formation is an immune response for the invasion of microbes. The purpose of this study is to examine the effect of zinc on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation of PMNs was measured by fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α in the culture supernatants from zinc-treated peripheral blood mononuclear cells (PBMCs) was measured by enzyme-linked immunosorbent assay (ELISA). Zinc itself did not have no effect on NET formation. However, the NET formation of PMNs was increased by culture supernatants from PBMCs treated with zinc. Also, the NET formation of PMNs was increased by recombinant porcine (rp) TNF-α. The production of TNF-α in PBMCs culture supernatants was shown to increase upon zinc treatments. These NET formations of PMNs increased by either culture supernatant from PBMCs treated with zinc or rpTNF-α were inhibited by treatment of anti-rpTNF-α polyclonal antibody (pAb). These results suggested that zinc has an immunostimulating effect on the NET formation of PMNs, which is mediated by TNF-α released from zinc-treated PBMCs. Therefore, zinc may play an important role for NET formation in the defense of porcine inflammatory diseases.

Cytokines Expression and Nitric Oxide Production under Induced Infection to Salmonella Typhimurium in Chicken Lines Divergently Selected for Cutaneous Hypersensitivity

Singh, Rani,Jain, Preeti,Pandey, N.K.,Saxena, V.K.,Saxena, M.,Singh, K.B.,Ahmed, K.A.,Singh, R.P. Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.7

In the present study, the impact of Salmonella Typhimurium on cell-mediated immunity (CMI) was investigated in 5 week-old immuno divergent broiler lines selected for the high and low response to phytohemagglutinin-P. The immune response was assessed in peripheral-blood mononuclear cells (PBMCs) induced with Salmonella Typhimurium at different time intervals (0 h, 0.5 h, 2 h, 4 h, 6 h, 12 h and 24 h). The differential mRNA expression patterns of IFN-${\gamma}$, IL-2 and iNOS were evaluated by quantitative real time PCR. In-vitro production of nitric oxide (NO) was also estimated in the culture supernatant and correlated with iNOS mRNA expression. Present study showed higher production of NO in the high cell-mediated line (HCMI) as compared to the low cell-mediated line (LCMI) upon stimulation with Salmonella Typhimurium. Correspondingly, higher mRNA expression of iNOS and IFN-${\gamma}$ were observed in high response birds (HCMI); but IL-2 was down regulated in this line compared to the low response birds (LCMI). Significantly (p<0.05) higher expression of iNOS, IFN-${\gamma}$ and higher production of NO in high line indicated that the selection for PHA-P response might be employed for increasing the immune competence against Salmonella Typhimurium in chicken flocks.

한우의 말초혈액 단핵세포 염증성 사이토카인에 대한 브로피리민의 효과 확인

김승창 ( Seungchang Kim ),이승환 ( Seung-Hwan Lee ),채한화 ( Han-Ha Chai ),김의형 ( Ui-Hyung Kim ),정기용 ( Ki-Yong Chung ),임다정 ( Dajeong Lim ),박종은 ( Jong-Eun Park ),조용민 ( Yongmin Cho ),최봉환 ( Bong-Hwan Choi ) 한국축산학회(구 한국동물자원과학회) 2017 동물자원연구 Vol.28 No.4

Bropirimine, a class of antineoplastic agents, is known as one of the potent immunomodulators and is currently under clinical development for the treatment of cancer. However, the effect of bropirimine on the cow remains unknown as a therapeutics agent. In this experiment, the effect of bropirimine in the peripheral blood mononuclear cells (PBMCs) stimulated by lipopolysaccharide (LPS) or concanavalin-A (Con-A) was examined. Jugular venous blood was collected from Korean Hanwoo calves and PBMCs were isolated. It was used to study the effect of bropirimine upon stimulation with LPS or Con-A for 72 hours. The expression pro-inflammatory cytokines like Tumor Necrosis Factor α (TNF-α) and Interferon γ (IFN-γ) were confirmed. Bropirimine significantly inhibited LPS- or Con-A-induced TNF-α and Con-A-induced IFN-γ in dose-dependent manner. Furthermore, Bropirimine inhibited TNF-α and Con-A mRNA expression at the transcription level. These results clearly indicated that bropirimine inhibited LPS or Con-A stimulated up-regulation of proinflammatory cytokines in a dose-dependent manner without conspicuous cytotoxicity. The bropirimine has potential to protect cow from LPS or Con-A induced endotoxin shock, possibly through inhibition of the production of proinflammatory cytokines. It suggesting that bropirimine may be a novel therapeutic agent for the prevention of inflammatory diseases. This result revealed specific features of the immune responses depending on the bropirimine compound and would help to knowledge of bovine immunity.

Anti-inflammatory effects of osthole in peripheral blood mononuclear cells from Hanwoo (Bos taurus coreanae)

김승창,이승환,채한화,김의형,정기용,장선식,최봉환 충남대학교 농업과학연구소 2019 Korean Journal of Agricultural Science Vol.46 No.3

Due to the ban on the use of antibiotics, interest has been increasing for the development of therapeutic agents to treat various diseases using natural resources. Osthole, a natural coumarin compound used in traditional Chinese medicines, exerts an anti-inflammatory effect, but its effects in cows remain unknown. In this study, the effect of osthole on lipopolysaccharide (LPS)- or concanavalin-A (Con-A)- stimulated peripheral blood mononuclear cells (PBMCs) was assessed. Jugular venous blood was collected from Korean calves, and PBMCs were isolated. They were then used to study the immune response of PBMCs to treatment with osthole and LPS or Con-A for 72 h by measuring inflammatory cytokines including tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). Osthole significantly inhibited the mRNA secretion of TNF-α and IFN-γ in a dose-dependent manner. Therefore, osthole inhibited LPS- or Con-A- induced TNF-α and Con-A-induced IFN-γ production significantly in dose-dependent manner. These results clearly suggest that osthole inhibited the LPS- or Con-A- stimulated upregulation of pro-inflammatory cytokines in a dose-dependent manner, without causing obvious cytotoxic effects. Osthole could also protect cows from LPS- or Con-A- induced endotoxin shock, possibly by inhibiting the production of pro-inflammatory cytokines, which suggests that osthole might be a novel therapeutic agent for the prevention of inflammatory diseases.

Effect of Surfactin on Growth Performance of Weaning Piglets in Combination with Bacillus subtilis BC1212

김명석,임종환,박병권,황윤환,송인배,박승춘,윤효인 한국임상수의학회 2009 한국임상수의학회지 Vol.26 No.2

The aim of this study is to investigate the effects of surfactin in combination with Bacillus subtilis BC1212 isolated from Korean soybean paste, on feed utilization and growth performance during 4 weeks in weaning piglets. Eighteen weaning piglets (Landrace × Yorkshire × Duroc; weighing 7.68±0.97 kg) were divided into control (n=9) and experimental groups (n=9). The treatments included a control group consisting of the basal diet with no additives (control) and an experimental group consisting of the basal diet supplemented with 1 g of surfactin C and 1.0×109 CFU of Bacillus subtilis BC1212/kg feed. Piglets fed Bacillus subtilis BC1212 increased in average daily weight gain and feed efficiency. In comparison with the control group, the fecal Bacillus subtilis were significantly increased and the fecal coliform bacteria were markedly reduced in the experimental group. In addition, Bacillus subtilis BC1212 had excellent acid and bile tolerance. The treatment of surfactin (50 μg ml-1) in lipopolysaccharide (LPS)-stimulated swine peripheral blood mononuclear cells (PBMCs) for 6 h showed a significant inhibitory effect on INF-γ, TNF-α and NO secretion (p<0.05) in comparison with LPS treatment alone but not on IL-10 secretion, with levels of secreted IL-10 similar to those secreted by PBMCs stimulated with LPS alone. Supplementation with surfactin in combination with Bacillus subtilis BC1212 in diets improved the ecosystem of gastrointestinal tract by increasing probiotic population and enhanced the systemic immune response in weaned piglets.