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>

Comparison of Allergic Parameters between Whey Protein Concentrate and Its Hydrolysate in Rat Basophilic Leukemia (RBL)-2H3 Cells

Hana Kim,Sung-Il Ahn,Jin-Woo Jhoo,Gur-Yoo Kim 한국축산식품학회 2018 한국축산식품학회지 Vol.38 No.4

This study was conducted to compare the anti-allergic effects of a whey protein concentrate (WPC) and WPC hydrolysate. WPC hydrolysate was prepared using enzymatic digestion for 8 h with trypsin and α-chymotrypsin, after which it was freezedried. The allergic parameters assessed in rat basophilic leukemia (RBL)-2H3 cells were degranulation and release of β-hexosaminidase, release of tumor necrosis factor (TNF)-α, and changes in the expression of IL-1β, IL-4, and IL-10 by real time polymerase chain reaction (PCR). During preparation of the WPC hydrolysate, hydrolysis increased rapidly from 0 to 10 min and then gradually increased slowly from 1 h onwards, achieving a final degree of hydrolysis of 78.50%. The SDS-PAGE analysis revealed a reduction in the intensity of several protein bands in the WPC hydrolysate compared to the WPC. IgEinduced β-hexosaminidase release from RBL-2H3 cells was decreased to a higher degree following treatment with the hydrolysate compared to WPC treatment. W500 (500 μg/mL WPC) showed the least inhibition of β-hexosaminidase release, but there was no significant difference between W500 and W1000 (1,000 μg/mL) (p<0.05). H1000 (1,000 μg/mL WPC hydrolysate) inhibited β-hexosaminidase release by 39%. Compared to the control, treatment with H1000 decreased TNF-α secretion to 11.87 pg/mL. The gene expression levels of IL-1β, IL-4, and IL-13 were all significantly decreased in hydrolysate (p<0.05). In the case of IL-1β and IL-4, the expression levels in W1000 treated cells were decreased by 73.67% and 65%, respectively, and that of IL-13 was decreased by 66.43% compared to the control.

Early Phase of UVB - induced GM - CSF Upregulation in Epithelial Cell Line is not Totally Dependent on IL - 1α

Park, Kyoung Chan,Kim, Kyu Han,Ahn, Jong Seong,Chung, Jin Ho,Youn, Jai Il,Whang, Ji Hwan,Youn, Sang Woong,Kim, Young Gull,Koh, Woo Seok,Jung, Hyun Chae 대한피부과학회 1997 Annals of Dermatology Vol.9 No.4

Backgrounds : It was demonstrated that ultraviolet(UV) B light induces the release of IL-la in cultured human epithelial cell line and augmentation of GM-CSF production by UVB is reported to be mediated by IL-1α in the murine keratinocyte cell line Pam 212. Objective : The purpose of this study was to evaluate the effects of UVB on kinetic profile of IL-1 and GM-CSF mRNA expression and to see whether synthesis of GM-CSF by UVB can be completely inhibited by blocking IL-1α mediated pathway. Method : We used a competitive RT-PCR for measuring cytokine gene expression in epithelial cell line after UV radiation. Results : The IL-1α mRNA increased as early as 1h after UV irradiation, and then decreased at 3h after the irradiation. Thereafter, the response of IL-1α mRNA was upregulated with a second peak at 6h after the UV irradiation. However, mRNA for GM-CSF increased at 1h after UV light exposure and anti-IL-1α antibodies could only partially inhibit UV-augmented GMCSF production. Conclusion : UVB induced GM-CSF production seemed to be mainly mediated by UVB induced IL-1α but these results suggest that UVB may also induce GM-CSF production through an IL-1α independent pathway.

생쥐의 B 세포에서 anti-CD40과 rIL-4로 유도된 싸이토카인 생산에 대한 자오가의 효과

성일창 ( Il Chang Sung ),김형환 ( Hyung Hwan Kim ),안덕균 ( Duk Kyun Ahn ),이용섭 ( Yong Sup Lee ),서영배 ( Young Bae Seo ),최호영 ( Ho Young Choi ) 대한본초학회 2003 大韓本草學會誌 Vol.18 No.2

N/A Objectives : In order to study the anti-allegy effect of water extract of Acanthopanacis senticosi Radix (ASR) on the B-cells from healthy Balblc mice. Methods : The cytotoxicity of ASR was measured with the murine normal lung fibroblast cells by modified SRB assay. And the murine splenic B-cells was stimulated with anti-CD40 mAb and rIL4. The various cytokines related with allergy were measured by flow-cytometry and by RT-PCR with electophoresis. Results : The anti-allegy effects to ASR were identified and observed. The cytotoxicity of ASR on mouse lung fibroblast cells showed no significant activities. ASR had inhibitory effect on CD23+, CD69+, and IgE expression by ASR with anti-CD40 mAb plus rL-4-stimulated murine splenic B-cells. ASR had inhibitory effect on cytokines (E-lb, IL-4, IL-6, IL-10, TNF-a, TGF-81, INF-Y) and transcript expression and IgE production by ASR with anti-CD40 mAb plus rIL-4-stimulated murine splenic B-cells. Conclusion : We concluded that ASR showed anti-allegy effect on murine splenic B-cells.

Characterization of IL-3, IL-5, GM-CSF - induced Eosinophils from Human CD34+ Cord Blood Cells

( Kwang Shin Lee ),( Kang Mo Ahn ),( Seung Yeon Nam ),( Dae Yeul Son ),( Se Chang Han ),( Man Yong Han ),( Sang Il Lee ) 대한 소아알레르기 호흡기학회(구 대한소아알레르기 및 호흡기학회) 1999 소아알레르기 및 호흡기학회지 Vol.9 No.4

제대혈 유래 인간 비만세포에서의 세포증식 및 히스타민 분비에 대한 Interleukin 9의 영향

안강모 ( Kang Mo Ahn ),이광신 ( Kwang Shin Lee ),신미용 ( Mi Yong Shin ),박화영 ( Hwa Young Park ),안연화 ( Yeon Hwa Ahn ),손대열 ( Dae Yeul Son ),이상일 ( Sang Il Lee ) 대한소아알레르기호흡기학회(구 대한소아알레르기 및 호흡기학회) 2002 소아알레르기 및 호흡기학회지 Vol.12 No.4

목적: Interleukin 9(IL-9)는 Th2 싸이토카인의 일종으로서 알레르기 염증반응의 병태생리에 관여하는 것으로 알려져 있다. 본 연구에서는 IL-9이 주요 알레르기 염증세포의 하나인 인간 비만세포에 어떠한 영향을 주는지를 알아보기 위하여 시행되었다. 방법:본 연구에서는 인간 제대혈에서 CD34 (+) 세포를 분리한 후 stem cell factor(SCF), IL-3, IL-6를 투여함으로써 비만세포를 선택적으로 배양하였다. 8주간 배양이 끝 Purpose:Interleukin-9(IL-9), one of Th2-type cytokines, might be important in the pathophysiology of allergic diseases. We investigated the effect of IL-9 on human mast cells by assessing cell proliferation and histamine release. Methods: Human umbilical

Antioxidant Activity of Yogurt Fermented at Low Temperature and Its Anti-inflammatory Effect on DSS-induced Colitis in Mice

Ji-Woo Yoon,Sung-Il Ahn,Jin-Woo Jhoo,Gur-Yoo Kim 한국축산식품학회 2019 한국축산식품학회지 Vol.39 No.1

This study was performed to evaluate the antioxidant activity of yogurt fermented at low temperature and the anti-inflammatory effect it has on induced colitis with 2.5% dextran sodium sulfate (DSS) in Balb/c mice. Yogurt premix were fermented with a commercial starter culture containing Lactobacillus acidophilus, Bifidobacterium lactis, Streptococcus thermophilus, and Lactobacillus delbrueckii subsp. bulgaricus at different temperatures: 22℃ (low fermentation temperature) for 27 h and 37℃ (general fermentation temperature) for 12 h. To measure antioxidant activity of yogurt samples, DPPH, ABTS+ and ferric reducing antioxidant potential (FRAP) assays were conducted. For animal experiments, inflammation was induced with 2.5% DSS in Balb/c mice. Yogurt fermented at low temperature showed higher antioxidant activity than that of the yogurt fermented at general temperature. In the inflammatory study, IL-6 (interleukin 6) was decreased and IL-4 and IL-10 increased significantly in DSS group with yogurt fermented at general temperature (DYG) and that with yogurt fermented at low temperature (DYL) compared to that in DSS-induced colitic mice (DC), especially DYL had higher concentration of cytokines IL-4, and IL-10 than DYG. MPO (myeloperoxidase) tended to decrease more in treatments with yogurt than DC. Additionally, yogurt fermented at low temperature had anti-inflammatory activity, although there was no significant difference with general temperature-fermented yogurt (p>0.05).

<i>In vivo</i> genotoxicity evaluation of lung cells from Fischer 344 rats following 28 days of inhalation exposure to MWCNTs, plus 28 days and 90 days post-exposure

Kim, Jin Sik,Sung, Jae Hyuck,Choi, Byung Gil,Ryu, Hyeon Yeol,Song, Kyung Seuk,Shin, Jae Hoon,Lee, Jong Seong,Hwang, Joo Hwan,Lee, Ji Hyun,Lee, Gun Ho,Jeon, Kisoo,Ahn, Kang Ho,Yu, Il Je Informa Healthcare 2014 INHALATION TOXICOLOGY Vol. No.

<P>Despite their useful physico-chemical properties, carbon nanotubes (CNTs) continue to cause concern over occupational and human health due to their structural similarity to asbestos. Thus, to evaluate the toxic and genotoxic effect of multi-wall carbon nanotubes (MWCNTs) on lung cells <I>in vivo</I>, eight-week-old rats were divided into four groups (each group = 25 animals), a fresh air control (0 mg/m<SUP>3</SUP>), low (0.17 mg/m<SUP>3</SUP>), middle (0.49 mg/m<SUP>3</SUP>), and high (0.96 mg/m<SUP>3</SUP>) dose group, and exposed to MWCNTs <I>via</I> nose-only inhalation 6 h per day, 5 days per week for 28 days. The count median length and geometric standard deviation for the MWCNTs determined by TEM were 330.18 and 1.72 nm, respectively, and the MWCNT diameters ranged from 10 to 15 nm. Lung cells were isolated from five male and five female rats in each group on day 0, day 28 (only from males) and day 90 following the 28-day exposure. The total number of animals used was 15 male and 10 female rats for each concentration group. To determine the genotoxicity of the MWCNTs, a single cell gel electrophoresis assay (Comet assay) was conducted on the rat lung cells. As a result of the exposure, the olive tail moments were found to be significantly higher (<I>p</I> < 0.05) in the male and female rats from all the exposed groups when compared with the fresh air control. In addition, the high-dose exposed male and middle and high-dose exposed female rats retained DNA damage, even 90 days post-exposure (<I>p</I> < 0.05). To investigate the mode of genotoxicity, the intracellular reactive oxygen species (ROS) levels and inflammatory cytokine levels (TNF-α, TGF- β, IL-1, IL-2, IL-4, IL-5, IL-10, IL-12 and IFN-γ) were also measured. For the male rats, the H<SUB>2</SUB>O<SUB>2</SUB> levels were significantly higher in the middle (0 days post-exposure) and high- (0 days and 28 days post-exposure) dose groups (<I>p</I> < 0.05). Conversely, the female rats showed no changes in the H<SUB>2</SUB>O<SUB>2</SUB> levels. The inflammatory cytokine levels in the bronchoalveolar lavage (BAL) fluid did not show any statistically significant difference. Interestingly, the short-length MWCNTs deposited in the lung cells were persistent at 90 days post-exposure. Thus, exposing lung cells to MWCNTs with a short tube length may induce genotoxicity.</P>

Antiviral and anti-inflammatory activity of budesonide against human rhinovirus infection mediated via autophagy activation

Kim, Seong-Ryeol,Song, Jae-Hyoung,Ahn, Jae-Hee,Lee, Geun-Shik,Ahn, Huijeong,Yoon, Sung-il,Kang, Seung Goo,Kim, Pyeung-Hyeun,Jeon, Sang-Min,Choi, Eun-Ji,Shin, Sooyoung,Cha, Younggil,Cho, Sungchan,Kim, Elsevier 2018 Antiviral research Vol.151 No.-

<P>Human rhinovirus (HRV) infection causes more than 80% of all common colds and is associated with severe complications in patients with asthma and chronic obstructive pulmonary disease. To identify antiviral drug against HRV infection, we screened 800 FDA-approved drugs and found budesonide as one of the possible drug candidates. Budesonide is a corticosteroid, which is commonly used to prevent exacerbation of asthma and symptoms of common cold. Budesonide specifically protects host cells from cytotoxicity following HRV infection, which depend on the expression of glucocorticoid receptor. Intranasal administration of budesonide lowered the pulmonary HRV load and the levels of IL-1 beta cytokine leading to decreased lung inflammation. Budesonide regulates IL-1 beta production following HRV infection independent of inflammasome activation. Instead, budesonide induces mitochondrial reactive oxygen species followed by activation of autophagy. Further, the inhibition of autophagy following chloroquine or bafilomycin Al treatment reduced the anti-viral effect of budesonide against HRV, suggesting that the antiviral activity of budesonide was mediated via autophagy. The results suggest that budesonide represents a promising antiviral and anti-inflammatory drug candidate for the treatment of human rhinovirus infection.</P>

반복적 관절내 출혈이 관절 활막과 연골 세포의 변화에 미치는 영향

유명철(Myung Chul Yoo),조윤제(Yoon Jae Cho),김강일(Kang Il Kim),전성욱(Sung Wook Chun),소동혁(Dong Hyuk So),조형준(Hyung Jun Cho),양형인(Hyung-In Yang),이상훈(Sang-Hoon Lee),이연아(Yeon-Ahn Lee) 대한정형외과학회 2008 대한정형외과학회지 Vol.43 No.3

목적: 혈액의 관절강내 반복주사 동물 모델을 이용하여, 혈우병성 관절염에서 활막의 변화와 연골 파괴에 대한 병리 기전과 경과 과정을 연구하였다. 대상 및 방법: 뉴질랜드 백색 토끼 수컷 20마리에 18주간 1주 3회씩 귀에서 자가 혈액을 채취하여 우측 슬관절에 1 ㎖를 주사하고, 좌측 슬관절에 생리식염수 1㎖를(대조군) 주사하였다. 11주 후와 18주 후에 양측 슬관절의 X선 촬영을 시행하였고, 활액막과 연골을 채취하여 조직검사를 시행하였다. 활막 세포 배양에서 RT PCR를 이용한 cytokine의 변화를 관찰 하였고, 연골 세포를 추출 배양하여 GAG 및 PGE₂, MMP-1,3 생성을 측정하였다. 결과: 11주째 육안적 변화는 없었으나, 우측 슬관절의 활액막은 조직학적으로 경한 증식반응과 형질세포 및 단핵세포의 활액막내 침착을 보이며 약간의 염증 소견을 나타냈다. 단순 방사선 소견상 특이 소견은 발견되지 않았다. 18주 후에는 육안적으로 우측 슬관절이 좌측에 비하여 부어 있었고, 단순 방사선학적 소견에서는 관절의 퇴행성 변화가 관찰 되었다. 병리학적 소견은 활막의 심한 증식과 염증세포의 침윤이 관찰되었다. 활막의 real-time RT PCR 시행결과 TNF-alpha, IL-1, MMP-1 mRNA 발현이 증가되었다. 연골세포 배양에서 대조군과 비교시 GAG 생성은 감소하고 PGE₂는 증가하였으며 MMP-1과 MMP-3는 변화가 없었다. 결론: 반복적인 관절 내 출혈은 활막세포를 자극하여 활막의 심한 증식과, proinflammatory cytokines의 생성을 증가시키며, 이는 연골 세포의 재생을 억제하고 연골세포의 염증을 증가시켜 연골의 대사기능이 억제되면서 퇴행성 변화를 촉진하여 점차적으로 관절염으로 이행되는 것으로 사료된다. Purpose: We designed this study to demonstrate the pathophysiology of hemophilic arthropathy (HA) by creating an animal model for determining the effect of repeated intraarticular bleeding in the synovium and articular cartilage. Materials and Methods: 20 normal male New Zealand white rabbits were used for this study. We injected 1 ㎖ of autologous blood from the ear vein of the rabbits into the right knee joint three timeds a week for 18 weeks, and we injected 1 ㎖ of normal saline into the left knee joint three times a week for 18 weeks as a control group. We examined the pathologic changes by microscopy and plain X-ray, and we determined the mRNA expression of proinflammatory cytokines in the synovium of the HA by performing real time RT-PCR at the 11<SUP>th</SUP> week and 18<SUP>th</SUP> week after starting blood-injection. We also examined the GAG and the PGE2 production in cultured chondrocytes that were extracted from the HA knees. Results: At the 11<SUP>th</SUP> week, after blood injection there were no remarkable gross changes in the HA knees and the control knee joints. At the 18<SUP>th</SUP> weeks, the experimental knee joints (HA knees) showed grossly swelling and degenerative changes by X-ray. The infiltration of inflammatory cells and the synovial proliferation in the HA knee joints were compared with that in the control knee joints by microscopic examination. The expressions of the mRNA of TNF-alpha, IL-1, MMP-1 and MMP-3 in the HA synovium were increased, as determined by real time RT-PCR, as compared with that in the control knee. In the cultured chondrocytes, the GAG production was decreased and the PGE2 was increased, but the MMP-1 and MMP-3 were not changed, as determined by ELISA. Conclusion: Our results showed that the GAG production of chondrocytes of the HA knees was decreased and there was increased PGE2, so that the cartilage degeneration by intra-articular bleeding was caused by the decreased metabolism of chondrocytes rather than by increased catabolism of the chondrocytes. We suggest that HA was associated with synovitis and cartilage degeneration, but decreased cartilage metabolism was the major mechanism of HA.