A Novel Small-Molecule Inhibitor Targeting the IL-6 Receptor β Subunit, Glycoprotein 130

Hong, Soon-Sun,Choi, Jung Ho,Lee, Sung Yoon,Park, Yeon-Hwa,Park, Kyung-Yeon,Lee, Joo Young,Kim, Juyoung,Gajulapati, Veeraswamy,Goo, Ja-Il,Singh, Sarbjit,Lee, Kyeong,Kim, Young-Kook,Im, So Hee,Ahn, Sun The American Association of Immunologists, Inc. 2015 JOURNAL OF IMMUNOLOGY Vol.195 No.1

<P>IL-6 is a major causative factor of inflammatory disease. Although IL-6 and its signaling pathways are promising targets, orally available small-molecule drugs specific for IL-6 have not been developed. To discover IL-6 antagonists, we screened our in-house chemical library and identified-LMT-28, a novel synthetic compound, as a candidate IL-6 blocker. The activity, mechanism of action, and direct molecular target of LMT-28 were investigated. A reporter gene assay showed that LMT-28 suppressed activation of STAT3 induced by IL-6, but not activation induced by leukemia inhibitory factor. In addition, LMT-28 downregulated IL-6-stimulated phosphorylation of STAT3, gp130, and JAK2 protein and substantially inhibited IL-6-dependent TF-1 cell proliferation. LMT-28 antagonized IL-6-induced TNF-alpha production in vivo. In pathologic models, oral administration of LMT-28 alleviated collagen-induced arthritis and acute pancreatitis in mice. Based on the observation of upstream IL-6 signal inhibition by LMT-28, we hypothesized IL-6, IL-6R alpha, or gp130 to be putative molecular targets. We subsequently demonstrated direct interaction of LMT-28 with gp130 and specific reduction of IL-6/IL-6R alpha complex binding to gp130 in the presence of LMT-28, which was measured by surface plasmon resonance analysis. Taken together, our data suggest that LMT-28 is a novel synthetic IL-6 inhibitor that functions through direct binding to gp130.</P>

Impact of interleukin-21 in the pathogenesis of primary Sjogren's syndrome: increased serum levels of interleukin-21 and its expression in the labial salivary glands

Kang, Kwi Young,Kim, Hyun-Ok,Kwok, Seung-Ki,Ju, Ji Hyeon,Park, Kyung-Su,Sun, Dong-Il,Jhun, Joo Yeon,Oh, Hye Jwa,Park, Sung-Hwan,Kim, Ho-Youn BioMed Central 2011 ARTHRITIS RESEARCH AND THERAPY Vol.13 No.5

<P><B>Introduction</B></P><P>Interleukin (IL)-21 is a cytokine that controls the functional activity of effector T helper cells and the differentiation of Th17 cells, and promotes B-cell differentiation. To test whether IL-21 participates in the pathogenesis of primary Sjögren's syndrome (SS), serum IL-21 level was measured and IL-21 expression in the labial salivary glands (LSG) was examined.</P><P><B>Methods</B></P><P>Serum IL-21 levels in 40 primary SS, 40 rheumatoid arthritis (RA), and 38 systemic lupus erythematosus (SLE) patients and 20 healthy controls were measured. Serum IL-21 levels of SS patients were assessed for correlations with laboratory data, including anti-nuclear antibody, anti-Ro/La antibodies, globulin, immunoglobulin (Ig) class, and IgG subclass. LSGs from 16 primary SS and 4 controls with sicca symptoms were evaluated for IL-21 and IL-21 receptor (IL-21R) expression by immunohistochemistry. Confocal microscopy was performed to further characterize the IL-21 positive cells.</P><P><B>Results</B></P><P>Primary SS patients had significantly higher serum IL-21 levels than controls, and these increments correlated positively with levels of IgG, IgG1. Serum IgG1 levels correlated with anti-Ro antibody titers. Immunohistochemical analyses showed that lymphocytic foci and the periductal area of the LSGs from SS patients expressed high levels of IL-21 and lower levels of IL-21R, whereas the control LSGs showed minimal expression of both antigens. The more the lymphocyte infiltrated, IL-21expression in LSGs showed a tendency to increase. Confocal microscopic analyses revealed that IL-21 expressing infiltrating lymphocytes in the LSGs of SS patients also expressed CXCR5.</P><P><B>Conclusions</B></P><P>Primary SS is associated with high serum IL-21 levels that correlate positively with serum IgG, especially IgG1, levels. The expression of IL-21 is increased as more lymphocytes infiltrated in LSGs. These observations suggest that IL-21 may play an important role in primary SS pathogenesis.</P>

Sequential evolution of IL-17 responses in the early period of allograft rejection

Sang Il Min,하종원,박정규,Jae Kyung Won,Yang Jin Park,민승기,김상준 생화학분자생물학회 2009 Experimental and molecular medicine Vol.41 No.10

In addition to CD4+CD25+Foxp3+ regulatory T (Treg) cells which protect against autoimmune tissue injury, IL-17-producing CD4+ T (Th17) cells have been recently described and shown to play a crucial role in autoimmune injury. It appears that there is a reciprocal developmental pathway between Th17 and Treg cells. Although IL-17 is known to be associated with allograft rejection, the cellular source of IL-17 and the nature of Th17 in the context of allograft rejection remain unknown. In the current study, the dynamics of Treg and IL-17-producing cells after syngeneic and allogeneic transplantation were examined using a wild-type murine cardiac transplantation model. Ly6G+ cells were found to produce IL-17 during the early postoperative period and CD8+ as well as CD4+ T cells were also found to produce IL-17 during alloimmune response. Graft-infiltrating Ly6G+, CD4+, and even CD8+ cells were found to express IL-17 highly compared to those in spleen. Although the frequencies of Th17 and Treg were found to gradually increase in both syngeneic and allogeneic recipients, Th17/Treg ratios were significantly higher in recipients with allograft rejection than in syngeneic recipients. In conclusion, IL-17 is produced by neutrophils during the early postoperative period and subsequently by Th17 and CD8+ T cells during allograft rejection. Th17/Treg imbalance is associated with the development of allograft rejection. This study would provide basic information on Th17 biology for future investigation in the field of transplantation.

Free Paper Session : Upper Gastrointestinal Tract 1 ; Prevalence And Risk Factors For Atrophic Gastritis And Intestinal Metaplasia

( Na Young Kim ),( Dong Ho Lee ),( Joo Sung Kim ),( Hyun Chae Jung ),( In Sung Song ),( Kyung Phil Kang ),( Jung Hoon Lee ),( Jae Il Chung ),( Hyun Cheul Choi ),( Taek Man Nam ),( Sang Hyup Lee ),( Yo 대한소화기학회 2007 SIDDS Vol.9 No.-

Background/Aims: The prevalence of gastric cancer and Helicobacter pylori (Hp) infection is high in Korea. This study was performed to evaluate the prevalence rate of atrophic gastritis (AG) and intestinal metaplasia (IM) and their risk factors in the aspect of Hp virulence factors, environmental and host factors in normal population. Methods: The subjects consisted of 389, 135 H. pylori-negative and 254 H. pylori-positive. AG and IM were scored histologically by the Sydney classification in the antrum and body, respectively. Prevalence rate and bacterial factors such as cagA, vacA m1, m2, and oipA; environmental factors such as smoking, alcohol drinking; host factors such as genetic polymorphisms for IL-IB-511, IL-IRN, TNF-A, IL-10-592, IL-10-819, IL-10-1082, IL-8-251, IL-6-572, GSTP1, and p53 codon 72 were evaluated. Risk factors were calculated by multiple logistic regression analysis. Results: The prevalence rate of AG increased from 25%, 0% in the age of 20s, 45% and 22% in the 40s and 50% and 35% in the over 70s in the antrum and body, respectively (p<0.001). In case of IM it increased from 11.1% and 6.4% in the 30s up to 43% and 43% in over 70s in the antrum and body, respectively, (p<0.001). The positive rates of AG and IM were significantly higher in the Hp-positive than in the Hp-negative subjects. Multivariate analysis showed that the risk factors for AG were Hp infection, age ≥60, cagA and vacA m1 positive. In case of IM the risk factors were Hp infection, age ≥60, smoking, spicy food, occupation (unemployed or non professional vs. professional), IL6-572 G carrier over C/C and IL10-592 C/A vs. A/A. Conclusions: The prevalence rate of AG and IM increased proportional to age. The most risk factor for AG and IM was Hp infection. Bacterial factors were important for AG but environmental and host factors were rather important in case of IM.

Selective addition of CXCR3+CCR4-CD4+ Th1 cells enhances generation of cytotoxic T cells by dendritic cells in vitro

Sung Hee Yoon,Sun Ok Yun,Jung Yong Park,Hee Yeun Won,Eun-Kyung Kim,Hyun-Jung Sohn,Hyun-Il Cho,김태규 생화학분자생물학회 2009 Experimental and molecular medicine Vol.41 No.3

Increasing importance is being given to the stimulation of Th1 response in cancer immunotherapy because its presence can shift the direction of adaptive immune responses toward protective immunity. Based on chemokine receptor expression, CXCR3+CCR4-CD4+ T cells as Th1-type cells were investigated its capacity in monocyte-derived dendritic cell (DC) maturation and polarization, and induction of antigen specific cytotoxic T lymphocytes (CTL) in vitro. The levels of IL-4, IL-5 and IL-10 were decreased to the basal level compared with high production of IFN-γ, TNF-α, and IL-2 in CXCR3+CCR4-CD4+ T cells stimulated with anti-CD3 and anti-CD28 antibodies. Co-incubation of activated CD4+ or CXCR3+CCR4-CD4+ T cells with DC (CD4+/DC or CXCR3+CD4+/DC, respectively) particularly up-regulated IL-12 and CD80 expression compared with DC matured with TNF-α and LPS (mDC). Although there was no significant difference between the effects of the CXCR3+CCR4-CD4+ and CD4+ T cells on DC phenotype expression, CXCR3+CD4+/DC in CTL culture were able to expand number of CD8+ T cells and increased frequencies of IFN-γ secreting cells and overall cytolytic activity against tumor antigen WT-1. These results demonstrated that the selective addition of CXCR3+ CCR4- CD4+ T cells to CTL cultures could enhance the induction of CTLs by DC in vitro, and implicated on a novel strategy for adoptive T cell therapy. Increasing importance is being given to the stimulation of Th1 response in cancer immunotherapy because its presence can shift the direction of adaptive immune responses toward protective immunity. Based on chemokine receptor expression, CXCR3+CCR4-CD4+ T cells as Th1-type cells were investigated its capacity in monocyte-derived dendritic cell (DC) maturation and polarization, and induction of antigen specific cytotoxic T lymphocytes (CTL) in vitro. The levels of IL-4, IL-5 and IL-10 were decreased to the basal level compared with high production of IFN-γ, TNF-α, and IL-2 in CXCR3+CCR4-CD4+ T cells stimulated with anti-CD3 and anti-CD28 antibodies. Co-incubation of activated CD4+ or CXCR3+CCR4-CD4+ T cells with DC (CD4+/DC or CXCR3+CD4+/DC, respectively) particularly up-regulated IL-12 and CD80 expression compared with DC matured with TNF-α and LPS (mDC). Although there was no significant difference between the effects of the CXCR3+CCR4-CD4+ and CD4+ T cells on DC phenotype expression, CXCR3+CD4+/DC in CTL culture were able to expand number of CD8+ T cells and increased frequencies of IFN-γ secreting cells and overall cytolytic activity against tumor antigen WT-1. These results demonstrated that the selective addition of CXCR3+ CCR4- CD4+ T cells to CTL cultures could enhance the induction of CTLs by DC in vitro, and implicated on a novel strategy for adoptive T cell therapy.

Interleukin-4와 히스타민이 알레르기성 비염과 비 알레르기성 비염에서 비점막 섬유아세포에 미치는 영향

김성완,조중생,박재경,차창일,김광일 대한이비인후과학회 1999 대한이비인후과학회지 두경부외과학 Vol.42 No.3

Solubility of Carbon Dioxide in Strongly Basic Ionic Liquid

Sung, Jun-Kyung,Kim, Sung-Hyun,Cheong, Min-Serk,Baek, Il-Hyun Korean Chemical Society 2010 Bulletin of the Korean Chemical Society Vol.31 No.10

For the purpose of developing advanced new absorbents for carbon dioxide, ionic liquids (ILs) are considered as alternative materials due to their superior properties to conventional organic solvents. Since low $CO_2$ solubility in ionic liquids is a major concern for their application as absorbents, it is essential to focus on improving $CO_2$ absorbing capability of ILs. In this paper, strongly basic ionic liquids, namely [$C_n$-mim]OPh (n = 2, 4, 6), have been synthesized and studied over a wide range of temperature and pressure changes. [$C_n$-mim]OPh can be easily synthesized from corresponding [$C_n$-mim]Cl and sodium phenoxide and has been found to be good $CO_2$ absorbents.

비용종 상피세포와 섬유아세포 동시배양 모델에서 시토카인의 발현

김성완,조중생,박재경,이건희,차창일 WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2007 東西醫學硏究所 論文集 Vol.2007 No.-

Background and Objectives: Nasal polyposis is a chronic inflammatory disease characterized histologically by edematous fluid with sparse fibrous cells, few mucous glands and proliferation of stromal and epithelial cells. There have been many studies done separately about epithelial cells, fibroblasts, and other inflammatory cells in nasal polyps. However, there is no usable study model for the cell-to-cell interaction. Therefore, we tried to make a new model for the study of nasal polyp by coculture of epithelial cells and fibroblasts. Subjects and Method: Nasal polyp was collected during endoscopic sinus surgery. Almost half of them were used for epithelial cell culture and the remaining portion was used for fibroblast culture in the same polyp. Using a transwell insert fibroblasts were placed in the bottom and epithelial cells were placed on the insert respectively. RNA were isolated both from cpithelial cells and fibroblasts of 12,24, and 48 hours' coculture. The expressions of IL-8,IL-6, and eotaxin from coculturcd cells were compared with cells that were cultured alone by semiquantitative RT-PCR. Results: IL-8 was highly expressed in fibroblasts than in epithelial cells, especially in cocultured cells. The expression of IL-6 in fibroblasts was much higher than in epithelial cells, especially in cocultured fibroblasts. Eotaxin is weakly expressed in fibroblasts but not in epithelial cells. Conclusion : These findings indicate that cocultured cells show different expression of the mRNA of various cytokines compared with cultured cells alone. And this coculture system may be more ideal than the mono-cell culture system for the study of cell-to-cell interactions and to reveal the pathogenesis of oasal polyposis.

위장관 ; Lipopolysaccharide로 유도된 HT-29 세포주의 염증에서 Lactobacillus rhamnosus GG의 항염증 작용과 기전

이상길 ( Sang Kil Lee ),양경민 ( Kyung Min Yang ),천재희 ( Jae Hee Cheon ),김태일 ( Tae Il Kim ),김원호 ( Won Ho Kim ) 대한소화기학회 2012 대한소화기학회지 Vol.60 No.2

Background/Aims: Probiotics are live non-pathogenic organisms that belong to the resident microflora, and confer health benefits by multiple mechanisms. Lactobacillus rhamnosus GG (LGG) is one of the probiotic bacteria that ameliorates intestinal injury and inflammation caused by various stimuli. We aimed to evaluate the anti-inflammatory effect and mechanism of LGG in lipopolysaccharide (LPS)-stimulated HT-29 cells. Methods: HT-29 cells were stimulated with interleukin (IL)-1β (2 ng/mL), tumor necrosis factor (TNF)-α (20 ng/mL), and LPS (20 μg/mL) in the presence or absence of LGG (107-109 colony forming units/mL). Production of the pro-inflammatory chemokine IL-8 was measured by ELISA and semi-quantitative PCR. Transcriptional activity of NF-κB-responsive gene was evaluated by luciferase assay with reporter gene. Toll-like receptor 4 (TLR4) mRNA expression was assessed by semi-quantitative PCR. The IκBα degradation was evaluated by western blot and intranuclear translocation of NF-κB was determined by western blot and immunofluorescence. Results: LGG did not affect the viability of HT-29 cells. Pretreatment of HT-29 cells with LGG significantly blocked TNF-α, and LPS induced IL-8 activation at both mRNA and protein level (p<0.05). Pretreatment of HT-29 cells with LGG attenuated LPS-induced NF-κB nuclear translocation and also blocked LPS-induced IκBα degradation. LGG also down-regulated TLR4 mRNA activated by LPS. Conclusions: LGG attenuates LPS induced inflammation, and this may be associated with TLR4/NF-κB down-regulation. (Korean J Gastroenterol 2012;60:86-93)

큰느타리버섯 균사체로 제조한 발효두부 추출물의 면역 활성

이상원(Sang-Won Lee),강종우(Jong-Woo Kang),김재용(Jae-Yong Kim),박경욱(Kyung-Wuk Park),박석규(Seok-Kyu Park),주옥수(Ok-Soo Joo),이성태(Sung-Tae Yee),서권일(Kwon-Il Seo) 한국식품영양과학회 2010 한국식품영양과학회지 Vol.39 No.1

두부의 기능성 및 저장성을 향상시킬 목적으로 큰느타리버섯 균사체를 이용한 발효두부를 제조하여 물과 메탄올로 추출하여 면역세포 활성에 미치는 효과를 조사하였다. 큰느타리버섯 균사체를 배양하기 위한 최적 배지는 PD broth 배지인 것을 확인하였으며, 큰타리버섯 균사체를 이용한 두부의 최적 발효기간은 7일 정도가 적당하였다. 큰느타리버섯 균사체를 이용하여 발효한 두부의 물 및 메탄올추출물은 0.01 ㎍/mL 농도 이상에서 비장세포의 증식을 유도하였으며, 이들 추출물은 IL-6, IFN-γ 분비를 유도하는 것으로 나타났다. 발효두부 물 추출물은 대조군에 비해 대식세포의 일산화질소 생산을 1 ㎍/mL 농도 이상에서 유의적으로 증가시키는 것을 알 수 있었으며, 메탄올 추출물은 10 ㎍/mL 농도 이상에서 그 생산을 증가시켰다. 발효두부 추출물들은 대식세포가 분비하는 IL-6, TNF-α, IL-1β 및 GM-CSF 분비량을 유의하게 증가시켰다. 따라서 큰느타리버섯 균사체로 발효한 두부는 기능성 두부로 개발이 가능하리라 생각된다. In order to improve the functional benefits and storage properties of soybean tofu, fermented tofu was developed using Pleurotus eryngii mycelia. The immune activities of water and methanol extracts of the tofu were investigated. The optimal medium for the growth of Pleurotus eryngii mycelia was PD broth medium and the optimal fermentation period for the tofu was 7 days. The water and methanol extracts of the fermented tofu induced the proliferation of spleen cells at above 0.01 ㎍/mL. The water extract increased IL-2, IFN-γ production, while the methanol extract increased IFN-γ synthesis. The water and methanol extracts of the fermented tofu induced the NO production in RAW264.7 macrophage cells at above 1 ㎍/mL and above 10 ㎍/mL concentration, respectively. The extracts also significantly increased the production of IL-6, TNF-α, IL-1β and GM-CSF in the cells. These results suggest that the tofu fermented with Pleurotus eryngii mycelia could be developed as a functional tofu.