IL-17 induces the production of IL-16 in rheumatoid arthritis

조미라,김경운,박미경,Hye-Joa Oh,주지현,Young-Gyu Cho,민준기,김성일,박성환,김호연,정영옥 생화학분자생물학회 2008 Experimental and molecular medicine Vol.40 No.2

The purpose of this study was to investigate the expression of IL-16 in the rheumatoid synovium and the role of inflammatory cytokines and Toll-like receptor (TLR) ligands in IL-16 production by fibroblastlike synoviocytes (FLS) of rheumatoid arthritis (RA) patients. Immunohistochemical staining was performed with a monoclonal antibody to IL-16 in synovial tissues from patients with RA and likewise in patients with osteoarthritis (OA). FLS were isolated from RA synovial tissues and stimulated with IL-15, IL-1β, IFN-γ, and IL-17. The IL-16 mRNA level was assessed by semiquantitative RT-PCR and real time (RT) PCR and a comparison was made between IL-16 mRNA levels produced by RA-FLS and OA-FLS. Production of IL-16 was identified by a western blot assay, and IL-16 production after stimulation by specific ligands of TLR2 and TLR4 was assessed by RT-PCR. While immunohistochemical staining demonstrated strong expression of IL-16 mRNA in synovial tissues from patients with RA, similar findings were not present in the OA group. Moreover, mRNA expression of IL-16 by RA-FLS increased after treatment with IL-17 but not with IL-15, IL-1β, and IFN-γ. Specifically, IL-17 increased IL-16 mRNA level by RA-FLS and peripheral blood mononuclear cells in a dose-dependent manner. However, IL-17 did not stimulate IL-16 production in OA-FLS. Peptidoglycan, a selective TLR2 ligand, also increased production of IL-16 by RA-FLS dosedependently, whereas LPS, a selective TLR4 ligand, had no such stimulatory effect. The results from our data demonstrate that IL-17 and TLR2 ligands stimulate the production of IL-16 by RA-FLS.

Anti-tumor Activity of Ethanol Extract of Androsace umbellata L. In Vitro and In Vivo

Eun-Kyung Ahn(안은경),Hye-Jin Ko,Jaeyeon Lee,Joa Sub Oh,Ok Pyo Zee 한국실험동물학회 2009 한국실험동물학회 학술발표대회 논문집 Vol.2009 No.-

Anti-obesity Effects of Ethanol Extract of Pinus Thunbergii Parl. In Vitro and in vivo

Eun-Kyung Ahn,Hye-Jin Ko,Jung A Lee,Seong Su Hong,Hyun Hwa Lee,Ju Young Shin,Joa Sub Oh 한국실험동물학회 2014 한국실험동물학회 학술발표대회 논문집 Vol.2014 No.2

Obesity aggravates the joint inflammation in a collagen-induced arthritis model through deviation to Th17 differentiation

전주연,윤보영,박미경,Hye-Joa Oh,Jae-Kyeong Byun,이선영,민준기,박성환,김호연,조미라 생화학분자생물학회 2012 Experimental and molecular medicine Vol.44 No.7

White fat cells secrete adipokines that induce inflammation and obesity has been reported to be characterized by high serum levels of inflammatory cytokines such as IL-6 and TNF-α. Rheumatoid arthritis (RA) is a prototype of inflammatory arthritis,but the relationship between RA and obesity is controversial. We made an obese inflammatory arthritis model: obese collagen-induced arthritis (CIA). C57BL/6 mice were fed a 60-kcal high fat diet (HFD) from the age of 4 weeks and they were immunized twice with type II collagen (CII). After immunization, the obese CIA mice showed higher arthritis index scores and histology scores and a more increased incidence of developing arthritis than did the lean CIA mice. After treatment with CII, mixed lymphocyte reaction also showed CII-specific response more intensely in the obese CIA mice than lean CIA. The anti-CII IgG and anti-CII IgG2a levels in the sera of the obese CIA mice were higher than those of the lean CIA mice. The number of Th17 cells was higher and the IL-17 mRNA expression of the splenocytes in the obese CIA mice was higher than that of the lean CIA mice. Obese CIA mice also showed high IL-17 expression on synovium in immunohistochemistry. Although obesity may not play a pathogenic role in initiating arthritis, it could play an important role in amplifying the inflammation of arthritis through the Th1/Th17 response. The obese CIA murine model will be an important tool when we investigate the effect of several therapeutic target molecules to treat RA.

IL-17-deficient allogeneic bone marrow transplantation prevents the induction of collagen-induced arthritis in DBA/1J mice

박민정,Mi-La Cho,Hyun-Sil Park,Hye-Joa Oh,윤보영,임정연,조미라,조석구 생화학분자생물학회 2012 Experimental and molecular medicine Vol.44 No.11

IL-17-producing CD4+ T cells (Th17) play important functions in autoimmune diseases and allograft rejection of solid organs. We examined the effects of IL 17 and its mechanism of action on arthritis in a murine collagen-induced arthritis (CIA) model using bone marrow transplantation (BMT) system. DBA/1J mice were administered a lethal radiation dose and then rescued with bone marrow derived from either wild-type (WT) or IL-17-/- mice on C57BL/6 background mice. CIA was induced after the bone marrow transplant, and disease progression was characterized. DBA/1J mice with CIA that received IL-17-/- donor bone marrow showed potently inhibited development and severity of clinical arthritis as compared with CIA mice that received WT bone marrow. Reduced secretion of the pro-inflammatory cytokines tumor necrosis factor-α,IL-1β, and IL-6, and collagen-specific T cell responses were observed in mice that received IL-17-/- bone marrow. IL-17 blockade also inhibited effector T cell proliferation by reciprocally regulating the Treg/Th17ratio. IL-17 blockade prevented joint destruction in mice with CIA. These findings suggest that CIA with BMT is a viable method of immunological manipulation and that IL-17 deficiency suppresses severe joint destruction and inflammation in CIA mice. There may be clinical benefits in blocking IL-17 and BMT in the treatment of rheumatoid arthritis.

Deficiency of $Foxp3^+$ Regulatory T Cells Exacerbates Autoimmune Arthritis by Altering the Synovial Proportions of $CD4^+$ T Cells and Dendritic Cells

Jang, Eun-Kyeong,Cho, Mi-La,Oh, Hye-Joa,Youn, Jee-Hee The Korean Association of Immunobiologists 2011 Immune Network Vol.11 No.5

Background: $CD4^+Fop3^+$ regulatory T cells (Tregs) are needed to maintain peripheral tolerance, but their role in the development of autoimmune arthritis is still debated. The present study was undertaken to investigate the mechanism by which Tregs influence autoimmune arthritis, using a mouse model entitled K/BxN. Methods: We generated Treg-deficient K/BxNsf mice by congenically crossing K/BxN mice with Foxp3 mutant scurfy mice. The arthritic symptoms of the mice were clinically and histopathologically examined. The proportions and activation of $CD4^+$ T cells and/or dendritic cells were assessed in the spleens, draining lymph nodes and synovial tissue of these mice. Results: K/BxNsf mice exhibited earlier onset and more aggressive progression of arthritis than their K/BxN littermates. In particular, bone destruction associated with the influx of numerous RANKL+ cells into synovia was very prominent. They also contained more memory phenotype $CD4^+$ T cells, more Th1 and Th2 cells, and fewer Th17 cells than their control counterparts. Plasmacytoid dendritic cells expressing high levels of CD86 and CD40 were elevated in the K/BxNsf synovia. Conclusion: We conclude that Tregs oppose the progression of arthritis by inhibiting the development of $RANKL^+$ cells, homeostatically proliferating $CD4^+$ T cells, Th1, Th2 and mature plasmacytoid dendritic cells, and by inhibiting their influx into joints.

Deficiency of Foxp3⁺ Regulatory T Cells Exacerbates Autoimmune Arthritis by Altering the Synovial Proportions of CD4⁺ T Cells and Dendritic Cells

Jang, Eun-Kyeong,Cho, Mi-La,Oh, Hye-Joa,Youn, Jee-Hee 대한면역학회 2011 Immune Network Vol.11 No.5

CD4⁺Fop3⁺ regulatory T cells (Tregs) are needed to maintain peripheral tolerance, but their role in the development of autoimmune arthritis is still debated. The present study was undertaken to investigate the mechanism by which Tregs influence autoimmune arthritis, using a mouse model entitled K/BxN. Methods: We generated Treg-deficient K/BxNsf mice by congenically crossing K/BxN mice with Foxp3 mutant scurfy mice. The arthritic symptoms of the mice were clinically and histopathologically examined. The proportions and activation of CD4⁺ T cells and/or dendritic cells were assessed in the spleens, draining lymph nodes and synovial tissue of these mice. Results: K/BxNsf mice exhibited earlier onset and more aggressive progression of arthritis than their K/BxN littermates. In particular, bone destruction associated with the influx of numerous RANKL+ cells into synovia was very prominent. They also contained more memory phenotype CD4⁺ T cells, more Th1 and Th2 cells, and fewer Th17 cells than their control counterparts. Plasmacytoid dendritic cells expressing high levels of CD86 and CD40 were elevated in the K/BxNsf synovia. Conclusion: We conclude that Tregs oppose the progression of arthritis by inhibiting the development of RANKL⁺ cells, homeostatically proliferating CD4⁺ T cells, Th1, Th2 and mature plasmacytoid dendritic cells, and by inhibiting their influx into joints.

Antagonism of VEGF-A-induced increase in vascular permeability by an integrin α3β1-Shp-1-cAMP/PKA pathway

Kim, Soo Hyeon,Cho, Young-Rak,Kim, Hyeon-Ju,Oh, Joa Sub,Ahn, Eun-Kyung,Ko, Hye-Jin,Hwang, Byung Joon,Lee, Seo-Jin,Cho, Yongwan,Kim, Yong Kee,Stetler-Stevenson, William G.,Seo, Dong-Wan American Society of Hematology 2012 Blood Vol.120 No.24

<B>Abstract</B><P>In cancer, VEGF-induced increase in vascular permeability results in increased interstitial pressure, reducing perfusion and increasing hypoxia, which reduce delivery of chemotherapeutic agents and increase resistance to ionizing radiation. Here, we show that both TIMP-2 and Ala + TIMP-2, a TIMP-2 mutant without matrix metalloproteinase inhibitory activity, antagonize the VEGF-A-induced increase in vascular permeability, both in vitro and in vivo. Like other agents known to preserve endothelial barrier function, TIMP-2 elevates cytosolic levels of cAMP and increases cytoskeletal-associated vascular endothelial cadherin in human microvascular endothelial cells. All of these effects are completely ablated by selective knockdown of integrin α3β1 expression, expression of a dominant negative protein tyrosine phosphatase Shp-1 mutant, administration of the protein tyrosine phosphatase inhibitor orthovanadate, or the adenylate cyclase inhibitor SQ22536. This TIMP-2-mediated inhibition of vascular permeability involves an integrin α3β1-Shp-1-cAMP/protein kinase A-dependent vascular endothelial cadherin cytoskeletal association, as evidenced by using siRNAs to integrin α3β1 and Shp-1, or treatment with Shp-1 inhibitor NSC87877 and protein kinase A inhibitor H89. Our results demonstrate the potential utility for TIMP-2 in cancer therapy through “normalization” of vascular permeability in addition to previously described antiangiogenic effects.</P>

산유자 잎 에탄올 추출물의 미백, 주름억제, 항염증 및 항산화 효능

이재연 ( Jae Yeon Lee ),안은경 ( Eun Kyung Ahn ),고혜진 ( Hye Jin Ko ),조영락 ( Young Rak Cho ),고운철 ( Woon Chul Ko ),정용환 ( Yong Hwan Jung ),최경민 ( Kyung Min Choi ),최미래 ( Mi Rae Choi ),오좌섭 ( Joa Sub Oh ) 한국응용생명화학회(구 한국농화학회) 2014 Journal of Applied Biological Chemistry (J. Appl. Vol.57 No.4

본 연구는 제주도 해변의 벌판에서 드물게 자라는 산유자(Xylosma congesta) 잎 에탄올 추출물의 화장품 기능성 소재로서의 활용 가능성을 알아보기 위하여 cell culture model 및 invitro assay system을 이용하여 미백, 주름개선, 항염증 및 항산화 활성을 분석하였다. 그 결과 산유자 잎 에탄올 추출물은 B16F10 세포에 세포독성 없이 효과적으로 α-MSH에 의해 유도된 melanin 합성을 억제함으로써 높은 미백 활성을 가지는 것이 관찰되었다. CCD-986SK 세포를 이용하여 산유자 잎 에탄올 추출물의 주름개선 활성 능력을 procollagen 합성시험을 통해 분석한 결과, 양성 대조군으로 사용한 TGF-β 만큼은 아니지만 농도별로 각각 120, 122%의 procollagen 합성을 증가시키는 것을 통하여 산유자 잎 에탄올 추출물의 주름개선 활성을 확인하였다. 또한 RAW 264.7 murine macrophage를 이용하여 NO 생성 억제능을 분석함으로써 산유자 잎 에탄올 추출물의 항염증 활성 정도를 측정한 결과, LPS에 의해 유도된 NO 생성이 산유자 잎 에탄올 추출물의 농도 의존적으로 감소하는 결과를 얻을 수 있었다. DPPH 법, ABTS 법, ORAC 법을 이용하여 항산화 활성을 분석한 결과, 50 μg/mL의 농도에서 DPPH및 ABTS 라디칼 소거능이 각각 75.2, 99.1% 증가하는 것이 확인 되었다. ORAC activity assay kit를 이용하여 항산화 활성을 측정한 결과 산유자 잎 에탄올 추출물의 농도에 따라 높은 항산화 활성이 관찰 되었고, DPPH와ABTS 라디칼 소거능 결과와 유사하게 50 μg/mL의 농도에서 가장 높은 항산화 활성(573.74±0.79 μM TE/g)을 나타내었다. 이러한 결과를 통하여 산유자 잎 에탄올 추출물이 높은 미백과 주름개선, 항염증 및 항산화 활성을 나타내는 것을 확인 할 수 있었으며, 기능성 화장품 소재로서의 활용가능성을 제시 할 수 있었다. In the present study, we investigated the biological activities of Xylosma congesta leaf ethanol extract (XCO) using a variety of in vitro and cell culture model systems for antimelanogenic, anti-wrinkle, anti-inflammatory and anti-oxidant activities. First, XCO markedly inhibited α-melanocyte stimulating hormone-stimulated melanin synthesis in B16F10 cells. Secondly, XCO marginally induced procollagen synthesis in CCD-986SK cells. Thirdly, XCO dose-dependently suppressed lipopolysaccharideinduced nitric oxide (NO) production in RAW 264.7 cells. XCO did not affect cell viability at different concentrations used in this study, indicating that XCO-mediated inhibition of melanin,procollagen and NO synthesis is not mediated by cytotoxicity. Finally, XCO was found to exert anti-oxidant effect. Taken together, these findings demonstrate for the first time that XCO possesses anti-melanogenic, anti-wrinkle, anti-inflammatory andanti-oxidant activities, and suggest further evaluation and development of XCO as a functional supplement or cosmetic that may be useful for whitening skin, reducing wrinkles and treating inflammatory responses.

Transforming growth factor β–transduced mesenchymal stem cells ameliorate experimental autoimmune arthritis through reciprocal regulation of Treg/Th17 cells and osteoclastogenesis

Park, Min‐,Jung,Park, Hyun‐,Sil,Cho, Mi‐,La,Oh, Hye‐,Joa,Cho, Young‐,Gue,Min, So‐,Youn,Chung, Byung‐,Ha,Lee, Jong‐,Wook,Kim, Ho‐,Youn,Cho, Seok&#x Wiley Subscription Services, Inc., A Wiley Company 2011 Vol.63 No.6

<P><B>Abstract</B></P><P><B>Objective</B></P><P>Bone marrow–derived mesenchymal stem cells (MSCs) can prevent various autoimmune diseases. We examined the therapeutic potential of transforming growth factor β (TGFβ)–transduced MSCs in experimental autoimmune arthritis, using an accepted animal model of collagen‐induced arthritis (CIA).</P><P><B>Methods</B></P><P>DBA/1J mice with CIA were treated with syngeneic TGFβ‐induced MSCs, whereas control mice received either vehicle or MSCs alone. Arthritis severity was assessed by clinical and histologic scoring. TGFβ‐transduced MSCs were tested for their immunosuppressive ability and differential regulation in mice with CIA. T cell responses to type II collagen were evaluated by determining proliferative capacity and cytokine levels. The effects of TGFβ‐transduced MSCs on osteoclast formation were analyzed in vitro and in vivo.</P><P><B>Results</B></P><P>Systemic infusion of syngeneic TGFβ‐transduced MSCs prevented arthritis development and reduced bone erosion and cartilage destruction. Treatment with TGFβ‐transduced MSCs potently suppressed type II collagen–specific T cell proliferation and down‐regulated proinflammatory cytokine production. These therapeutic effects were associated with an increase in type II collagen–specific CD4+FoxP3+ Treg cells and inhibition of Th17 cell formation in the peritoneal cavity and spleen. Furthermore, TGFβ‐transduced MSCs inhibited osteoclast differentiation.</P><P><B>Conclusion</B></P><P>TGFβ‐transduced MSCs suppressed the development of autoimmune arthritis and joint inflammation. These data suggest that enhancing the immunomodulatory activity of MSCs and modulating T cell–mediated immunity using gene‐modified MSCs may be a gateway for new therapeutic approaches to clinical rheumatoid arthritis.</P>