• IMMUNE NETWORK
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• 제11권6호
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• pp.383-389
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• 2011

Background: EY-6 is one of the newly synthesized indoledione derivatives to induce tumor cell-specific cell death. In this study, we investigated the mechanism of immunological death induced by EY-6 at mouse colon cancer cell as well as at the normal immune cell represented by dendritic cell. Methods: C57BL/6 mouse syngeneic colon cancer cell MC38 was treated with EY-6, and analyzed by MTT for viability test, flow cytometry for confirming surface expressing molecules and ELISA for detection of cytokine secretion. Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC. Results: EY-6 killed the MC38 tumor cells in a dose dependent manner (25, 50 and $100{\mu}M$) with carleticulin induction. And EY-6 induced the secretion of IFN-${\gamma}$ but not of TNF-${\alpha}$ from the MC38 tumor cells. EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation. The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation). Conclusion: Data indicate that the EY-6 induced tumor cell specific and immunological cell death by modulation of tumor cell phenotype and cytokine secretion favoring induction of specific immunity eliminating tumor cells.

• Biomolecules & Therapeutics
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• 제19권1호
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• pp.77-83
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• 2011

Sulforaphane is an isothiocyanate found in cruciferous vegetables that has been reported to be an effective cancer preventive agent inducing growth arrest and/or cell death in cancer cells of various organs. This paper reports that sulforaphane exerts immunomodulatory activity on the MHC-restricted antigen presenting function. Sulforaphane efficiently increased the class II-restricted presentation of an exogenous antigen, ovalbumin (OVA), in both dendritic cells (DCs) and peritoneal macrophages in vitro. The class II-restricted OVA presentation-enhancing activity of sulforaphane was also confirmed using mice that had been injected with sulforaphane followed by soluble OVA. On the other hand, sulforaphane did not affect the class I-restricted presentation of exogenous OVA at concentrations that increase the class II-restricted antigen presentation. At a high concentration ($20\;{\mu}M$), sulforaphane inhibited the class I-restricted presentation of exogenous OVA. Sulforaphane did not affect the phagocytic activity of the DCs, and the cell surface expression of total H-$2K^b$, B7-1, B7-2 and CD54 molecules, even though it increased the expression of I-$A^b$ molecules to a barely discernable level. These results show that sulforaphane increases the class II-restricted antigen presenting function preferentially, and might provide a novel insight into the mechanisms of the anti-cancer effects of sulforaphane.

• 생명과학회지
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• 제27권4호
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• pp.482-499
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• 2017

Allergy 질환은 지난 십여년 동안 개발도상국을 포함해서 전 세계적으로 증가하고 있다. Allergy 염증 반응은 수지상 세포와 같은 항원제시 세포에 의한 allergy 항원섭취를 시작으로 하여 Th2 면역 반응에 의해서 일어난다. 장내 미생물은 신체의 대사나 생리적 기능을 조절하고, 생애 초기의 면역 체계 성숙과 일생 동안 면역 체계 항상성 및 상피세포 총체성에 기여한다. Bifidobacteria는 Th1/Th2 balance에 strain-specific 한 면역 자극 특성을 가지며, TSLP와 IgE 발현을 억제 시키고 Flg과 FoxP3 발현을 촉진 시켜 allergy를 완화시킨다. 또한 Unmethylated CpG motif ODN은 B 세포와 수지상 세포의 TLR9에 의해 인식 되어 선천성과 적응성 면역 반응을 유도하고, Clostridium butyricum에 의해서 생산된 butyrate는 수지상 세포의 anti-inflammatory 유전자의 발현을 유도하기 위해 GPR109a signaling pathway를 활성화시키고, GPR43 활성화를 통하여 tTreg 세포 proliferation을 직접 자극하거나 HADC 활성을 억제시켜 Foxp3 gene intronic enhancer의 histone H3 acetylation을 통해 naive $CD4^+$ T 세포를 pTreg 세포로 분화시킨다.

• IMMUNE NETWORK
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• 제23권5호
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• pp.41.1-41.21
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• 2023

CD4 and CD8 T cells are key players in the immune response against both pathogenic infections and cancer. CD4 T cells provide help to CD8 T cells via multiple mechanisms, including licensing dendritic cells (DCs), co-stimulation, and cytokine production. During acute infection and vaccination, CD4 T cell help is important for the development of CD8 T cell memory. However, during chronic viral infection and cancer, CD4 helper T cells are critical for the sustained effector CD8 T cell response, through a variety of mechanisms. In this review, we focus on T cell responses in conditions of chronic Ag stimulation, such as chronic viral infection and cancer. In particular, we address the significant role of CD4 T cell help in promoting effector CD8 T cell responses, emerging techniques that can be utilized to further our understanding of how these interactions may take place in the context of tertiary lymphoid structures, and how this key information can be harnessed for therapeutic utility against cancer.

• Biomolecules & Therapeutics
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• 제20권4호
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• pp.399-405
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• 2012

Silver nanoparticles (AgNPs) are widely used nanoparticles and they are mainly used in antibacterial and personal care products. In this study, we evaluated the effect of AgNPs on cell death induction in the murine dendritic cell line DC2.4. DC2.4 cells exposed to AgNPs showed a marked decrease in cell viability and an induction of lactate dehydrogenase (LDH) leakage in a time- and dose-dependent manner. In addition, AgNPs promoted reactive oxygen species (ROS)-dependent apoptosis and AgNP-induced ROS triggered a decrease in mitochondrial membrane potential. The activation of the intracellular signal transduction pathway was also observed in cells cultured with AgNPs. Taken together, our data demonstrate that AgNPs are able to induce a cytotoxic effect in DCs through ROS generation. This study provides important information about the safety of AgNPs that may help in guiding the development of nanotechnology applications.

• 대한한의학회지
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• 제43권2호
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• pp.1-7
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• 2022

Objectives: Bee venom (BV) is a widely used therapy in Traditional East Asian Medicine (TEAM). We previously reported that BV was clinically effective for treating Parkinson's disease, that phospholipase A₂ (PLA₂) was the main component of BV, and that it induced regulatory T cells (Tregs) by binding CD206 on dendritic cells (DCs). Therefore, we aimed to reconfirm our findings in human blood samples and investigate the relationship between CD206⁺ DCs and clinical syndrome differentiation in TEAM. Methods: We surveyed 100 subjects with questionnaires on cold-heat patternization and obtained their blood samples. The obtained human peripheral blood monocytes (hPBMCs) were washed with phosphate-buffered saline (PBS). After resuspension with ex vivo media, numbers of cells were counted. Tregs were counted after culturing the samples in a 37℃ CO₂ incubator for 72 h. Results: We divided the subjects into a relatively high CD206⁺ group or a relatively low CD206⁺ group. The heat factor scores of high CD206⁺ group were significantly higher than that of low CD206⁺ group (high vs low: 239.2 ± 54.1 vs 208.4 ± 55.1, p=0.023). After culturing with PLA₂, Tregs increased in the high CD206⁺ group but decreased in the low CD206⁺ group. Conclusion: In this study, we reconfirm that CD206⁺ DCs induced Treg differentiation by incubating human blood samples with PLA₂ and that they showed an association with syndrome differentiation, especially with heat patterns, in TEAM. A heat pattern in TEAM might be one indication for PLA₂ therapy because its score was elevated in the high CD206⁺ group.

• 한국식품과학회지
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• 제50권4호
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• pp.444-450
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• 2018

본 연구는 꿀벌 꽃가루 추출물(BPW)의 면역 활성에 관하여 알아보기 위하여, 선천면역계의 대표적인 수지상세포와 후천면역계의 대표적인 비장세포에 BPW를 처리 하여 면역세포의 활성능을 관찰하였다. 수지상 세포에 BPW를 처리하여 세포 생존율, 산화질소(II)와 사이토카인($TNF-{\alpha}$, IL-6과 $IL-1{\beta}$) 분비능과 세포 표면분자를 관찰 하였다. 세포 생존율은 수지상 세포에 BPW를 처리하였을 때, 세포 독성을 일으키지 않았으며 주요 면역 활성 인자인 산화질소(II) 분비능을 관찰한 결과, 농도 의존적으로 증가하는 것을 확인하였다. 또한 사이토카인의 분비능을 관찰한 결과, $TNF-{\alpha}$, IL-6과 $IL-1{\beta}$의 함량이 농도 의존적으로 증가하는 것으로 관찰되었다. 또한 활성화된 면역세포의 세포 표면에서 발현되는 CD80과 CD86의 발현과 항원제시에 밀접한 관련이 있는 MHC class I, II의 발현이 유의적으로 증가하는 것으로 관찰되었다. 또한 후천면역에서 중요한 역할을 수행하는 면역 T 세포가 다량 분포하는 지라 세포를 분리하여 BPW를 처리 하였을 때 Th1 세포가 분비하는 사이토카인의 함량이 농도 의존적으로 증가되는 것으로 나타났다. 이러한 결과로 미루어 볼 때 BPW는 선천면역뿐만 아니라 후천면역에 관여하는 다양한 면역세포의 활성화에 직간접적으로 관여하는 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제23권8호
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• pp.1159-1166
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• 2013

Orientia tsutsugamushi, a gram-negative bacterium, causes severe acute febrile illness in humans. Despite this danger, the route of infection, infectivity, and protective mechanisms of the host's immune response to O. tsutsugamushi are unclear. Dendritic cells (DCs) are one of the most important cell types in bridging the innate and adaptive immune responses. In this study, we observed that O. tsutsugamushi infects and replicates in monocyte-derived DCs (MODCs). During infection and replication, the expressions of the cytokines IL-12 and TNF-${\alpha}$, as well as the co-stimulatory molecules CD80, CD83, CD86, and CD40, were increased in MODCs. When O. tsutsugamushi-treated MODCs were co-cultured with autologous $CD4^+$ T cells, they enhanced production of IFN-${\gamma}$, a major Th1 cytokine. Collectively, our results show that O. tsutsugamushi can replicate in MODCs and can simultaneously induce MODC maturation and increase proinflammatory cytokine levels in MODCs that subsequently activate $CD4^+$ T cells.

• IMMUNE NETWORK
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• 제5권3호
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• pp.163-171
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• 2005

Background: To perform the successful dendritic cell-based cancer immunotherapy one of the main issues to be solved is the source of antigen for DC pulsing. Limitations occur by using auto-tumor lysate due to the difficulties obtaining enough tumor tissue(s) quantitatively as well as qualitatively. In this study the possibility of allogeneic tumor cell lysate as a DC pulsing antigen has been tested in mouse melanoma pulmonary me tastasis model. Methods: B16F10 melanoma cells $(1{\timeS}10^5/mouse)$ were inoculated intra venously into the C57BL/6 mouse. Therapeutic DCs were cultured from the bone marrow myeloid lineage cells with GM-CSF and IL-4 (1,000 U/ml each) for 7 days and pulsed with lysate of either autologous B16F10 (B-DC), allogeneic K1735 (C3H/He origin; K-DC) or CloneM3 (DBA2 origin; C-DC) melanoma cells for 18 hrs. Pulsed-DCs $(1{\times}10^6/mouse)_{[CGP1]}$ were injected i.p. twice with one week interval starting from the day 1 after tumor cell inoculation. Results: Without observable toxicity, allogeneic tumor cell lysate pulsed-DC induced the significantly better anti-tumor response (tumor scale: $2.7{\pm}0.3,\;0.7{\pm}0.3\;and\;0.3{\pm}0.2$ for saline, B-DC and C-DC treated group, respectively). Along with increased tumor specific lymphocyte proliferations, induction of IFN-${\gamma}$ secretion against both auto- and allo-tumor cell lysates was observed from the DC treated mice. (w/B16F10-lysate: $44.97{\pm}10.31,\;1787.94{\pm}131.18,\;1257.15{\pm}48.27$, w/CloneM3 lysate: 0, $1591.13{\pm}1.83,\;1460.47{\pm}86.05pg/ml$ for saline, B-DC and C-DC treated group, respectively) Natural killer cell activity was also increased in the mice treated with tumor cell lysate pulsed-DC ($8.9{\pm}_{[CGP2]}0.1,\;11.6{\pm}0.8\;and\;12.6{\pm}0.7%$ specific NK activity for saline, B-DC and C-DC treated group, respectively). Conclusion: Conclusively, promising data were obtained that allogeneic-tumor cell lysate can be used as a tumor antigen for DC-based cancer immunotherapy.

• IMMUNE NETWORK
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• 제10권4호
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• pp.135-143
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• 2010

Background: Cordyceps militarys water extract (CME) has been reported to exert antitumor and immunomodulatory activities in vivo and in vitro. However, the therapeutic mechanism has not yet been elucidated. In this study, we examined the effects of CME on the antigen presenting function of antigen presenting cells (APCs). Methods: Dendritic cells (DCs) were cultured in the presence of CME, and then allowed to phagocytose microspheres containing ovalbumin (OVA). After washing and fixing the efficacy of OVA, peptide presentation by DCs were evaluated using CD8 and CD4 T cells. Also, we confirmed the protein levels of proinflammatory cytokines through western blot analysis. Results: CME enhanced both MHC class I and class II-restricted presentation of OVA in DCs. In addition, the expression of both MHC class I and II molecules was enhanced, but there was no changes in the phagocytic activity of exogenous OVA. Furthermore, CME induced the protein levels of iNOS, COX-2, proinflammatory cytokines, and nuclear p65 in a concentration-dependent manner, as determined by western blot. Conclusion: These results provide an understanding of the mechanism of the immuno-enhancing activity of CME on the induction of MHC-restricted antigen presentation in relation to their actions on APCs.