류마티스관절염 환자의 활액에서 형광 리포터 생쥐 시스템을 이용한 인터루킨-12/인터루킨-23 p40 생성 유도물질의 발굴
Screening of Interleukin-12/interleukin-23 p40 Inducers in Rheumatoid Synovial Fluids by Fluorescence Reporter Mouse System
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류마티스관절염은 주로 가동관절에 만성적 염증 소견을 보이는 자가면역질환으로 그 발병기전이 잘 알 려져 있지 않다. 본 연구에서는 류마티스관절염에 걸린 관절조직에서 활성화된 병적 신호전달체계를 규명하기 위 한 노력의 일환으로, 류마티스관절염 또는 골관절염 환자의 무릎 관절에서 분리한 활액 속에 포함된 물질이 전염 증성 사이토카인인 인터루킨(IL)-12와 IL-23 생성을 유도할 수 있는지를 조사하였다. IL-12/IL-23 p40 형광 리포터 생쥐의 복강에서 분리한 대식세포를 활액으로 자극한 후, CD11b+ 세포 중에서 노 란형광을 띤 세포를 유세포분석기로 분석하여 IL-12/IL-23 p40의 생성을 유도하는 물질이 포함된 활액을 스크리 닝하였다. 이들 유도물질이 관절조직 속의 활막세포에서 Toll-like receptors (TLRs)를 통하여 신호를 보내 이들 사 이토카인의 생성을 유도할 가능성을 조사하기 위하여, 활막조직에서 분리한 활막세포에서 TLR-2와 TLR-4의 발 현 양상을 RT-PCR 방법으로 조사하였다. 10명의 류마티스관절염 환자 중 한명의 활액이 양성반응을 보였으며, 10명의 골관절염 환자의 활액은 모두 음 성반응을 보였다. 류마티스관절염 환자의 활막세포는 TLR-2와 TLR-4를 모두 발현하고 있었고, LPS 자극에 의하 여 TLR-2의 발현은 감소하고 TLR-4의 발현은 증가하는 변화를 보였다. 그러므로 본 연구결과는 류마티스관절염 환자의 일부는 관절조직에서 전염증물질로 알려진 IL-12와 IL-23의 생성되어 염증반응을 일으키며 이 과정을 TLR-2와 TLR-4가 매개할 가능성이 있음을 제시한다.
Abstract
Although rheumatoid arthritis has been known to be a common autoimmune disease characterized by chronic inflammation mainly evident in diarthrodial joints, its pathogenesis remains to be clarified. In the present study, to investigate the pathogenic signaling system taken place in the rheumatoid joints, we assessed whether synovial fluid obtained from patients with rheumatoid arthritis contains inducers for proinflammatory cytokines such as interleukin (IL)-12 and IL-23. Peritoneal macrophages isolated from IL-12/IL-23 p40-YFP reporter mice were stimulated with synovial fluid, followed by flow cytometry to screen CD11b+ and YFP-expressing cells, reflective of IL-12/IL-23 p40-producing macrophages. The expression levels of Toll-like receptor (TLR)-2 and -4, which have a potential to mediate IL-12/IL- 23 p40 induction, were determined in synovial cells obtained from a patient with rheumatoid arthritis by RT-PCR analyses. One out of 10 synovial fluid from rheumatoid arthritis patients induced IL-12/IL-23 p40 expression, while all of 10 synovial fluid from osteoarthritis patients did not. Synoviocytes constitutively expressed Toll-like receptor (TLR)-2 and -4 which are candidate receptors for IL-12/IL-23 inducers. Upon LPS stimulation, the levels of TLR-2 and -4 were downregulated and upregulated, respectively. Taken together, these results suggest that some patients with rheumatoid arthritis elicit synovitis through TLR-2- and -4-mediated induction of proinflammatory cytokines IL-12 and IL-23.
참고문헌 (22)
- "Interleukin-23 promotes a distinct CD4 T cell activationstate characterized by the production of interleukin-17.", 278, pp.1910~1914 (2003)
- "Toll-like receptor signaling.", 4, pp.499~511 (2004)
- "Novel signal transduction pathway utilized by extracellular HSP70.", 277, pp.15028~15034 (2002)
- "Cutting Edge: IL-23 cross-regulates IL-12 production in T cell-dependent experimental colitis", 177, pp.2760~2764 (2006)
- "The immunologic homunculus in rheumatoid arthritis.", 42, pp.2499~506 (1999)
- "Interleukin-23 rather than interleukin-12 is the critical cytokine for autoimmune inflammation of the brain", 421, pp.744~748 (2003)
- "Cytokine reporter mouse system for screening novelIL12p40-inducing compounds.", 20, pp.288~296 (2005)
- "Blockade of IL-12 during the induction of collagen-induced arthritis (CIA) markedly attenuates the severity of the arthritis.", 111, pp.377~383 (1998)
- "Reduced incidenceand severity of collagen-induced arthritis in interleukin-12-deficient mice.", 26, pp.2933~2938 (1996)
- "Divergent pro- and antiinflammatory roles for IL-23 andIL-12 in jointautoimmune inflammation.", 198, pp.1951~1957 (2003)
- "The molecular basis of T helper 1 andhelper 2 cell differentiation.", 10, pp.542~550 (2000)
- "Heat shock protein60 is a putative endogenous ligand of the toll-like receptor-4 complex.", 164, pp.558~561 (2000)
- "Visualization of IL-12", 23 p40 in vivo reveals immunostimulatorydendritic cell migrants that promote Th1 differentiation., 2006., pp.177, 1618~1627
- "Identification of small heat shock protein B8 (HSP22) as a novel TLR4 ligand and potential involvement in the pathogenesis of rheumatoid arthritis", 176, pp.7021~7027 (2006)
- "Toll-like receptors control activation of adaptiveimmune responses.", 2, pp.947~950 (2001)
- "Fibrinogen stimulatesmacrophage chemokine secretion through toll-like receptor4.", 167, pp.2887~2894 (2001)
- "Differential roles of TLR2and TLR4 in recognition of Gram-negative and Gram-positivebacterial cell wall components.", 11, pp.443~451 (1999)
- "Oligosaccharides of hyaluronan activate dendritic cells viatoll-like receptor 4.", 195, pp.99~111 (2002)
- "Interleukin-12 and the regulation of innate resistanceand adaptive immunity.", 3, pp.133~146 (2003)
- "Accelerated collagen-induced arthritis in IFN-gamma receptor-deficient mice.", 158, pp.5507~5513 (1997)
- "Lessons for the pathogenesis of rheumatoid arthritisacquired from experimental animal models.", 25, pp.53~66 (2005)
- "New cyclooxygenase-2 inhibitor DFU regulatesvascular endothelial growth factor expression in rheumatoidsynoviocytes.", 96, pp.219~224 (2005)