骨芽細胞によるCX3CL1の発現とその調節: 関節リウマチの病態形成への関与
CX3CL1 (フラクタルカイン) は膜結合型のケモカインであり細胞遊走活性のほかに細胞接着活性をも合わせ持っており関節リウマチ (RA) の病態にも関与している.RAの骨破壊において骨芽細胞 (OB) および破骨細胞の役割は重要である.この骨芽細胞におけるCX3CL1の発現調節機構を解析した.RAおよび変形性関節症 (OA) 患者の骨組織から骨芽細胞を分離培養してCX3CL1の産生ならびにmRNA発現をELISAおよび定量real time PCR測定した.正常コントロールは正常ヒトの外傷性骨関節手術時に分離した.RA由来OB (RAOB) からのCX3CL1産生は無刺激およびtumor n...
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| Published in | 昭和医学会雑誌 Vol. 68; no. 2; pp. 98 - 105 |
|---|---|
| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | Japanese |
| Published |
昭和大学学士会
28.04.2008
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0037-4342 2185-0976 |
| DOI | 10.14930/jsma1939.68.98 |
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| Abstract | CX3CL1 (フラクタルカイン) は膜結合型のケモカインであり細胞遊走活性のほかに細胞接着活性をも合わせ持っており関節リウマチ (RA) の病態にも関与している.RAの骨破壊において骨芽細胞 (OB) および破骨細胞の役割は重要である.この骨芽細胞におけるCX3CL1の発現調節機構を解析した.RAおよび変形性関節症 (OA) 患者の骨組織から骨芽細胞を分離培養してCX3CL1の産生ならびにmRNA発現をELISAおよび定量real time PCR測定した.正常コントロールは正常ヒトの外傷性骨関節手術時に分離した.RA由来OB (RAOB) からのCX3CL1産生は無刺激およびtumor necrosis factor alpha (TNF-α) , interferon gamma (IFN-γ) の刺激においてもほとんど産生はみられなかったが, TNF-αとIFN-γの共刺激により著明に産生増強が認められた.この発現は免疫染色ならびにフローサイトメトリーにおいても確認された.このOBからのCX3CL1産生はOAOBあるいはnormal OBに比してRAOBにおいて増強していた.このCX3CL1はmRNAレベルにおいても同様にRAOBにおいて高発現であった.サイトカインの共刺激による産生増強の機序を明らかにする目的で細胞内シグナル伝達因子の関与を検討した.RAOBによるnuclear factor kappa B (NF-κB) のmRNA発現はそれぞれのサイトカイン単独に比して増強していた.さらにこのNF-κBを阻害するpyrrolidineの添加によりCX3CL1の産生および発現は有意に低下した.またこのNF-κBに対するsiRNAをトランスフェクションし阻害することによりCX3CL1の産生および発現は有意に低下した.以上の結果は骨芽細胞がCX3CL1の重要な発現細胞でありその発現調節は炎症性サイトカインとNF-κBにより制御されていた. |
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| AbstractList | CX3CL1 (フラクタルカイン) は膜結合型のケモカインであり細胞遊走活性のほかに細胞接着活性をも合わせ持っており関節リウマチ (RA) の病態にも関与している.RAの骨破壊において骨芽細胞 (OB) および破骨細胞の役割は重要である.この骨芽細胞におけるCX3CL1の発現調節機構を解析した.RAおよび変形性関節症 (OA) 患者の骨組織から骨芽細胞を分離培養してCX3CL1の産生ならびにmRNA発現をELISAおよび定量real time PCR測定した.正常コントロールは正常ヒトの外傷性骨関節手術時に分離した.RA由来OB (RAOB) からのCX3CL1産生は無刺激およびtumor necrosis factor alpha (TNF-α) , interferon gamma (IFN-γ) の刺激においてもほとんど産生はみられなかったが, TNF-αとIFN-γの共刺激により著明に産生増強が認められた.この発現は免疫染色ならびにフローサイトメトリーにおいても確認された.このOBからのCX3CL1産生はOAOBあるいはnormal OBに比してRAOBにおいて増強していた.このCX3CL1はmRNAレベルにおいても同様にRAOBにおいて高発現であった.サイトカインの共刺激による産生増強の機序を明らかにする目的で細胞内シグナル伝達因子の関与を検討した.RAOBによるnuclear factor kappa B (NF-κB) のmRNA発現はそれぞれのサイトカイン単独に比して増強していた.さらにこのNF-κBを阻害するpyrrolidineの添加によりCX3CL1の産生および発現は有意に低下した.またこのNF-κBに対するsiRNAをトランスフェクションし阻害することによりCX3CL1の産生および発現は有意に低下した.以上の結果は骨芽細胞がCX3CL1の重要な発現細胞でありその発現調節は炎症性サイトカインとNF-κBにより制御されていた. |
| Author | 手塚, 正一 笠間, 毅 松倉, 聡 若林, 邦伸 磯崎, 健男 足立, 満 小田井, 剛 |
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| References | 14) Hasegawa M, Sato S, Echigo T, et al: Up regulated expression of fractalkine/CX3CL1 and CX3CR1 in patients with systemic sclerosis. Ann Rheum Dis 64: 21-28, 2005. 15) Yajima N, Kasama T, Isozaki T, et al: Elevated levels of soluble fractalkine in active systemic lupus erythematosus: Potential involvement in neuropsychiatric manifestations. Arthritis Rheum 52: 1670-1675, 2005. 20) Beck GC, Yard BA, Breedijk AJ, et al: Release of CXC-chemokines by human lung microvascular endothelial cells (LMVEC) compared with macrovascular umbilical vein endothelial cells. Clin Exp Immunol 118 : 298-303, 1999. 9) Kasama T, Isozaki T, Odai T, et al: Expression of angiopoietin-1 in osteoblasts and its inhibition by tumor necrosis factor-alpha and interferon-gamma. Transl Res 149: 265-273, 2007. 13) Blaschke S, Koziolek M, Schwarz A, et al: Promflammatory role of fractalkine (CX3CL1) in rheumatoid arthritis. J Rheumatol 30 : 1918-1927. 2003. 8) Lisignoli G, Toneguzzi S, Pozzi C, et al: Proinflammatory cytokines and chemokine production and expression by human osteoblasts isolated from patients with rheumatoid arthritis and osteoarthritis. J Rheumatol 26: 791-799, 1999. 26) Nanki T, Urasaki Y, Imai T, et al: Inhibition of fractalkine ameliorates murine collagen-induced arthritis. J Immunol 173: 7010-7016, 2004. 1) Kunkel SL, Lukacs N, Kasama T, et al: The role of chemokines in inflammatory joint disease. J Leukoc Biol 59 : 6-12, 1996. 3) Kasama T, Strieter RM, Lukacs NW, et al: Inter-leukin-10 expression and chemokine regulation during the evolution of murine type II collageninduced arthritis. J Clin Invest 95 : 2868-2876, 1995. 11) Umehara H, Bloom E, Okazaki T, et al: Fractalkine and vascular injury. Trends Immunol 22 : 602-607, 2001. 4) Kasama T: Collagen-induced arthritis: a useful model for studying the kinetics and regulation of the cytokine network in arthritis development. In Recent Research Developments in Immunology. vol. 5 (Ed by Pandalai SG) pp. 295-311, Research Signpost, Kerala, 2004. 18) Matsunawa M, Isozaki T, Odai T, et al: Increased serum levels of soluble fractalkine (CX3CL1) correlate with disease activity in rheumatoid vasculitis. Arthritis Rheum 54: 3408-3416, 2006. 5) Udagawa N, Kotake S, Kamatani N, et al: The molecular mechanism of osteoclastogenesis in rheumatoid arthritis. Arthritis Res 4 : 281-289, 2002. 16) Ruth JH, Volin MV, Haines GK, 3rd, et al: Fractalkine, a novel chemokine in rheumatoid arthritis and in rat adjuvant-induced arthritis. Arthritis Rheum 44: 1568-1581, 2001. 29) Lisignoli G, Toneguzzi S, Grassi F, et al: Different chemokines are expressed in human arthritic bone biopsies: IFN-gamma and IL-6 differently modulate IL-8, MCP-1 and rantes production by arthritic osteoblasts. Cytokine 20 : 231-238, 2002. 28) Ahn SY, Cho CH, Park KG, et al: Tumor necrosis factor-alpha induces fractalkine expression preferentially in arterial endothelial cells and mithramycin A suppresses TNF-alpha-induced fractalkine expression. Am J Pathol 164: 1663-1672, 2004. 12) Chen S, Bacon KB, Li L, et al: In vivo inhibition of CC and CX3C chemokine-induced leukocyte infiltration and attenuation of glomerulonephritis in Wistar-Kyoto (WKY) rats by vMIP-II. J Exp Med 188: 193-198, 1998. 27) Chandrasekar B, Mummidi S, Perla RP, et al: Fractalkine (CX3CL1) stimulated by nuclear factor kappaB (NF-kappaB) -dependent inflammatory signals induces aortic smooth muscle cell proliferation through an autocrine pathway. Biochem J 373 : 547-558, 2003. 24) Yoshida H, Imaizumi T, Fujimoto K, et al: Synergistic stimulation, by tumor necrosis factor-alpha and interferon-gamma of fractalkine expression in human astrocytes. Neurosci Lett 303 : 132-136, 2001. 25) Volin MV, Woods JM, Amin MA, et al: Fractalkine : a novel angiogenic chemokine in rheumatoid arthritis. Am J Pathol 159 : 1521-1530, 2001. 22) Ludwig A, Berkhout T, Moores K, et al: Fractalkine is expressed by smooth muscle cells in response to IFN-gamma and TNF-alpha and is modulated by metalloproteinase activity. J Immunol 168: 604-612, 2002. 30) van Roon JA, van Roy JL, Duits A, et al: Proinflammatory cytokine production and cartilage damage due to rheumatoid synovial T helper-1 activation is inhibited by interleukin-4. Ann Rheum Dis 54 : 836-840, 1995. 6) Zhu JF, Valente AJ, Lorenzo JA, et al: Expression of monocyte chemoattractant protein 1 in human osteoblastic cells stimulated by proinflam-matory mediators. J Bone Miner Res 9 : 1123-1130, 1994. 2) Choy EHS and Panayi GS: Cytokine pathways and joint inflammation in rheumatoid arthritis. N Engl J Med 344 : 907-916, 2001. 23) Sugaya M, Nakamura K, Mitsui H, et al: Human keratinocytes express fractalkine/CX3CL1. J Dermatol Sci 31: 179-187, 2003. 7) Kurokouchi K, Kambe F, Yasukawa K, et al: TNF-alpha increases expression of IL-6 and ICAM-1 genes through activation of NF-kappaB in osteoblast-like R0517/2.8 cells. J Bone Miner Res 13 : 1290-1299, 1998. 19) Bazan JF, Bacon KB, Hardiman G, et al: A new class of membrane-bound chemokine with a CX3C motif. Nature 385 : 640-644, 1997. 21) Lucas AD, Chadwick N, Warren BF, et al: The transmembrane form of the CX3CL1 chemokine fractalkine is expressed predominantly by epithelial cells in vivo. Am J Pathol 158 : 855-866, 2001. 17) Nanki T, Imai T, Nagasaka K, et al: Migration of CX3CR1-positive T cells producing type 1 cytokines and cytotoxic molecules into the synovium of patients with rheumatoid arthritis. Arthritis Rheum 46 : 2878-2883, 2002. 10) Imai T, Hieshima K, Haskell C, et al: Identification and molecular characterization of fractalkine receptor CX3CR1, which mediates both leukocyte migration and adhesion. Cell 91 : 521-530, 1997. |
| References_xml | – reference: 20) Beck GC, Yard BA, Breedijk AJ, et al: Release of CXC-chemokines by human lung microvascular endothelial cells (LMVEC) compared with macrovascular umbilical vein endothelial cells. Clin Exp Immunol 118 : 298-303, 1999. – reference: 24) Yoshida H, Imaizumi T, Fujimoto K, et al: Synergistic stimulation, by tumor necrosis factor-alpha and interferon-gamma of fractalkine expression in human astrocytes. Neurosci Lett 303 : 132-136, 2001. – reference: 17) Nanki T, Imai T, Nagasaka K, et al: Migration of CX3CR1-positive T cells producing type 1 cytokines and cytotoxic molecules into the synovium of patients with rheumatoid arthritis. Arthritis Rheum 46 : 2878-2883, 2002. – reference: 28) Ahn SY, Cho CH, Park KG, et al: Tumor necrosis factor-alpha induces fractalkine expression preferentially in arterial endothelial cells and mithramycin A suppresses TNF-alpha-induced fractalkine expression. Am J Pathol 164: 1663-1672, 2004. – reference: 5) Udagawa N, Kotake S, Kamatani N, et al: The molecular mechanism of osteoclastogenesis in rheumatoid arthritis. Arthritis Res 4 : 281-289, 2002. – reference: 1) Kunkel SL, Lukacs N, Kasama T, et al: The role of chemokines in inflammatory joint disease. J Leukoc Biol 59 : 6-12, 1996. – reference: 22) Ludwig A, Berkhout T, Moores K, et al: Fractalkine is expressed by smooth muscle cells in response to IFN-gamma and TNF-alpha and is modulated by metalloproteinase activity. J Immunol 168: 604-612, 2002. – reference: 10) Imai T, Hieshima K, Haskell C, et al: Identification and molecular characterization of fractalkine receptor CX3CR1, which mediates both leukocyte migration and adhesion. Cell 91 : 521-530, 1997. – reference: 23) Sugaya M, Nakamura K, Mitsui H, et al: Human keratinocytes express fractalkine/CX3CL1. J Dermatol Sci 31: 179-187, 2003. – reference: 16) Ruth JH, Volin MV, Haines GK, 3rd, et al: Fractalkine, a novel chemokine in rheumatoid arthritis and in rat adjuvant-induced arthritis. Arthritis Rheum 44: 1568-1581, 2001. – reference: 13) Blaschke S, Koziolek M, Schwarz A, et al: Promflammatory role of fractalkine (CX3CL1) in rheumatoid arthritis. J Rheumatol 30 : 1918-1927. 2003. – reference: 15) Yajima N, Kasama T, Isozaki T, et al: Elevated levels of soluble fractalkine in active systemic lupus erythematosus: Potential involvement in neuropsychiatric manifestations. Arthritis Rheum 52: 1670-1675, 2005. – reference: 27) Chandrasekar B, Mummidi S, Perla RP, et al: Fractalkine (CX3CL1) stimulated by nuclear factor kappaB (NF-kappaB) -dependent inflammatory signals induces aortic smooth muscle cell proliferation through an autocrine pathway. Biochem J 373 : 547-558, 2003. – reference: 25) Volin MV, Woods JM, Amin MA, et al: Fractalkine : a novel angiogenic chemokine in rheumatoid arthritis. Am J Pathol 159 : 1521-1530, 2001. – reference: 19) Bazan JF, Bacon KB, Hardiman G, et al: A new class of membrane-bound chemokine with a CX3C motif. Nature 385 : 640-644, 1997. – reference: 18) Matsunawa M, Isozaki T, Odai T, et al: Increased serum levels of soluble fractalkine (CX3CL1) correlate with disease activity in rheumatoid vasculitis. Arthritis Rheum 54: 3408-3416, 2006. – reference: 3) Kasama T, Strieter RM, Lukacs NW, et al: Inter-leukin-10 expression and chemokine regulation during the evolution of murine type II collageninduced arthritis. J Clin Invest 95 : 2868-2876, 1995. – reference: 29) Lisignoli G, Toneguzzi S, Grassi F, et al: Different chemokines are expressed in human arthritic bone biopsies: IFN-gamma and IL-6 differently modulate IL-8, MCP-1 and rantes production by arthritic osteoblasts. Cytokine 20 : 231-238, 2002. – reference: 30) van Roon JA, van Roy JL, Duits A, et al: Proinflammatory cytokine production and cartilage damage due to rheumatoid synovial T helper-1 activation is inhibited by interleukin-4. Ann Rheum Dis 54 : 836-840, 1995. – reference: 4) Kasama T: Collagen-induced arthritis: a useful model for studying the kinetics and regulation of the cytokine network in arthritis development. In Recent Research Developments in Immunology. vol. 5 (Ed by Pandalai SG) pp. 295-311, Research Signpost, Kerala, 2004. – reference: 21) Lucas AD, Chadwick N, Warren BF, et al: The transmembrane form of the CX3CL1 chemokine fractalkine is expressed predominantly by epithelial cells in vivo. Am J Pathol 158 : 855-866, 2001. – reference: 2) Choy EHS and Panayi GS: Cytokine pathways and joint inflammation in rheumatoid arthritis. N Engl J Med 344 : 907-916, 2001. – reference: 6) Zhu JF, Valente AJ, Lorenzo JA, et al: Expression of monocyte chemoattractant protein 1 in human osteoblastic cells stimulated by proinflam-matory mediators. J Bone Miner Res 9 : 1123-1130, 1994. – reference: 14) Hasegawa M, Sato S, Echigo T, et al: Up regulated expression of fractalkine/CX3CL1 and CX3CR1 in patients with systemic sclerosis. Ann Rheum Dis 64: 21-28, 2005. – reference: 26) Nanki T, Urasaki Y, Imai T, et al: Inhibition of fractalkine ameliorates murine collagen-induced arthritis. J Immunol 173: 7010-7016, 2004. – reference: 9) Kasama T, Isozaki T, Odai T, et al: Expression of angiopoietin-1 in osteoblasts and its inhibition by tumor necrosis factor-alpha and interferon-gamma. Transl Res 149: 265-273, 2007. – reference: 7) Kurokouchi K, Kambe F, Yasukawa K, et al: TNF-alpha increases expression of IL-6 and ICAM-1 genes through activation of NF-kappaB in osteoblast-like R0517/2.8 cells. J Bone Miner Res 13 : 1290-1299, 1998. – reference: 12) Chen S, Bacon KB, Li L, et al: In vivo inhibition of CC and CX3C chemokine-induced leukocyte infiltration and attenuation of glomerulonephritis in Wistar-Kyoto (WKY) rats by vMIP-II. J Exp Med 188: 193-198, 1998. – reference: 11) Umehara H, Bloom E, Okazaki T, et al: Fractalkine and vascular injury. Trends Immunol 22 : 602-607, 2001. – reference: 8) Lisignoli G, Toneguzzi S, Pozzi C, et al: Proinflammatory cytokines and chemokine production and expression by human osteoblasts isolated from patients with rheumatoid arthritis and osteoarthritis. J Rheumatol 26: 791-799, 1999. |
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| Snippet | CX3CL1 (フラクタルカイン) は膜結合型のケモカインであり細胞遊走活性のほかに細胞接着活性をも合わせ持っており関節リウマチ (RA) の病態にも関与している.RAの骨破... |
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| Title | 骨芽細胞によるCX3CL1の発現とその調節: 関節リウマチの病態形成への関与 |
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