Please use this identifier to cite or link to this item:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28259
Full metadata record
???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
---|---|---|
dc.contributor.advisor | 林琬琬(Wan-Wan Lin) | |
dc.contributor.author | Ying-Cing Lin | en |
dc.contributor.author | 林盈慶 | zh_TW |
dc.date.accessioned | 2021-06-13T00:03:48Z | - |
dc.date.available | 2012-08-08 | |
dc.date.copyright | 2007-08-08 | |
dc.date.issued | 2007 | |
dc.date.submitted | 2007-07-30 | |
dc.identifier.citation | Akira, S., Uematsu, S., and Takeuchi, O. (2006). Pathogen recognition and innate immunity. Cell 124, 783-801.
Alexopoulou, L., Holt, A.C., Medzhitov, R., and Flavell, R.A. (2001). Recognition of double-stranded RNA and activation of NF-kappaB by Toll-like receptor 3. Nature 413, 732-738. Arndt, P.G., Suzuki, N., Avdi, N.J., Malcolm, K.C., and Worthen, G.S. (2004). Lipopolysaccharide-induced c-Jun NH2-terminal kinase activation in human neutrophils: Role of phosphatidylinositol 3-kinase and Syk-mediated pathways. J Biol Chem. 279, 10883-10891. Bennett, B.L. (2006). c-Jun N-terminal kinase-dependent mechanisms in respiratory disease. Eur Respir J. 28, 651-661. Berton, G., Mocsai, A., and Lowell, C.A. (2005). Src and Syk kinases: key regulators of phagocytic cell activation. Trends Immunol. 26, 208-214. Beutler, B., Jiang, Z., Georgel, P., Crozat, K., Croker, B., Rutschmann, S., Du, X., and Hoebe, K. (2006). Genetic analysis of host resistance: Toll-like receptor signaling and immunity at large. Annu Rev Immunol. 24, 353-389. Bianchi, M., Ulrich, P., Bloom, O., Meistrell, M., Zimmerman, G.A., Schmidtmayerova, H., Bukrinsky, M., Donnelley, T., Bucala, R., Sherry, B., et al. (1995). An inhibitor of macrophage arginine transport and nitric-oxide production (CNI-1493) prevents acute-inflammation and endotoxin lethality. Mol Med. 1, 254-266. Chen, C.W., Lee, S.T., Wu, W.T., Fu, W.M., Ho, F.M., and Lin, W.W. (2003). Signal transduction for inhibition of inducible nitric oxide synthase and cyclooxygenase-2 induction by capsaicin and related analogs in macrophages. Br J Pharmacol. 140, 1077-1087. Chen, J.C., Ho, F.M., Pei-Dawn Lee, C., Chen, C.P., Jeng, K.-C.G., Hsu, H.B., Lee, S.T., Wen Tung, W., and Lin, W.W. (2005). Inhibition of iNOS gene expression by quercetin is mediated by the inhibition of IkappaB kinase, NF-kappaB and STAT1, and depends on heme oxygenase-1 induction in mouse BV-2 microglia. Eur J Pharmacol. 521, 9-20. Choe, J., Kelker, M.S., and Wilson, I.A. (2005). Crystal structure of human Toll-like receptor 3 (TLR3) ectodomain. Science 309, 581-585. Chu, W.M., Ostertag, D., Li, Z.-W., Chang, L., Chen, Y., Hu, Y., Williams, B., Perrault, J., and Karin, M. (1999). JNK2 and IKKbeta are required for activating the innate response to viral infection. Immunity 11, 721-731. Chung, J.Y., Park, Y.C., Ye, H., and Wu, H. (2002). All TRAFs are not created equal: common and distinct molecular mechanisms of TRAF-mediated signal transduction. J Cell Sci. 115, 679-688. Costello, P.S., Turner, M., Walters, A.E., Cunningham, C.N., Downward, J., Bauer, P., and Tybulewicz, V.L.J. (1996). Critical role for the tyrosine kinase Syk in signaling through the high affinity IgE receptor of mast cells. Oncogene. 13, 2595-2605. Couture, C., Williams, S., Gauthier, N., Tailor, P., and Mustelin, T. (1997). Role of Tyr518 and Tyr519 in the regulation of catalytic activity and substrate phosphorylation by Syk protein-tyrosine kinase. Eur J Biochem. 246, 447-451. Covert, M.W., Leung, T.H., Gaston, J.E., and Baltimore, D. (2005). Achieving stability of lipopolysaccharide-induced NF-kappaB activation. Science 309, 1854-1857. Crowley, M.T., Costello, P.S., Fitzer-Attas, C.J., Turner, M., Meng, F., Lowell, C., Tybulewicz, V.L., and DeFranco, A.L. (1997). A critical role for Syk in signal transduction and phagocytosis mediated by Fcgamma receptors on macrophages. J Exp Med. 186, 1027-1039. Cusson-Hermance, N., Khurana, S., Lee, T.H., Fitzgerald, K.A., and Kelliher, M.A. (2005). Rip1 mediates the TRIF-dependent Toll-like receptor 3- and 4-induced NF-kappaB activation but does not contribute to interferon regulatory factor 3 activation. J Biol Chem. 280, 36560-36566. Dong, C., Davis, R.J., and Flavell, R.A. (2002). MAK kinase in the immune response. Annu Rev Immunol. 20, 55-72. Ely, K.R., and Li, C. (2002). Structurally adaptive hot spots at a protein interaction interface on TRAF3. J Mol Recognit. 15, 286-290. Fitzgerald, K.A. (2003). IKKepsi and TBK1 are essential components of the IRF3 signaling pathway. Nature Immunol. 4, 491-496. Fodor, S., Jakus, Z., and Mocsai, A. (2006). ITAM-based signaling beyond the adaptive immune response. Immunol Lett. 104, 29-37. Gay, N.J., Gangloff, M., and Weber, A.N.R. (2006). Toll-like receptors as molecular switches. Nat Rev Immunol. 6, 693-698. Gotoh, Y., Oishi, K., Shibata, H., Yamagiwa, A., Isagawa, T., Nishimura, T., Goyama, E., Takahashi, M., Mukai, H., and Ono, Y. (2004). Protein kinase PKN1 associates with TRAF2 and is involved in TRAF2-NF-kappaB signaling pathway. Biochem Biophys Res Commun. 314, 688-694. Hacker, H., and Karin, M. (2006). Regulation and function of IKK and IKK-related kinases. Sci STKE 357, re13-. Hacker, H., Redecke, V., Blagoev, B., Kratchmarova, I., Hsu, L.-C., Wang, G.G., Kamps, M.P., Raz, E., Wagner, H., Hacker, G.., Mann, M., and Karin, M. (2006). Specificity in Toll-like receptor signalling through distinct effector functions of TRAF3 and TRAF6. Nature 439, 204-207. Hamerman, J.A., and Lanier, L.L. (2006). Inhibition of immune responses by ITAM-bearing receptors, Sci STKE. 320, re1- Hamerman, J.A., Tchao, N.K., Lowell, C.A., and Lanier, L.L. (2005). Enhanced Toll-like receptor responses in the absence of signaling adaptor DAP12. Nat Immunol. 6, 579-586. Hemmi, H., Takeuchi, O., Kawai, T., Kaisho, T., Sato, S., Sanjo, H., Matsumoto, M., Hoshino, K., Wagner, H., Takeda, K., and Akira, S. (2000). A Toll-like receptor recognizes bacterial DNA. Nature 408, 740-745. Hibi, M., Lin, A., Smeal, T., Minden, A., and Karin, M. (1993). Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. Genes Dev. 7, 2135-2148. Hoebe, K. (2003). Identification of Lps2 as a key transducer of MyD88-independent TIR signalling. Nature 424, 743-748. Hoebe, K., and Beutler, B. (2006). TRAF3: a new component of the TLR-signaling apparatus. Trends Mol Med. 12, 187-189. Iwasaki, A., and Medzhitov, R. (2004). Toll-like receptor control of the adaptive immune responses. Nat Immunol. 5, 987-995. Kaminska, B. (2005). MAPK signalling pathways as molecular targets for anti-inflammatory therapy--from molecular mechanisms to therapeutic benefits. Biochim Biophys Acta. 1754, 253-262. Karin, M., and Greten, F.R. (2005). NF-kappaB: Linking inflammation and immunity to cancer development and progression. Nat Rev Immunol. 5, 749-759. Kathryn, R., and Ely, C.L. (2002). Structurally adaptive hot spots at a protein interaction interface on TRAF3. J Mol Recognit. 15, 286-290. Kawai, T., Adachi, O., Ogawa, T., Takeda, K., and Akira, S. (1999). Unresponsiveness of MyD88-deficient mice to endotoxin. Immunity 11, 115-122. Kawai, T., and Akira, S. (2005). Pathogen recognition with Toll-like receptors. Curr Opin Immunol. 17, 338-344. Kawai, T., and Akira, S. (2006). TLR signaling. Cell Death Differ 13, 816-825. Kawai, T., and Akira, S. (2007). TLR signaling. Semin Immunol. 19, 24-32. Kawai, T., Takeuchi, O., Fujita, T., Inoue, J.-i., Muhlradt, P.F., Sato, S., Hoshino, K., and Akira, S. (2001). Lipopolysaccharide stimulates the MyD88-independent pathway and results in activation of IFN-regulatory factor 3 and the expression of a subset of lipopolysaccharide-inducible genes. J Immunol. 167, 5887-5894. Kawakami, Y., Kitaura, J., Yao, L., McHenry, R.W., Kawakami, Y., Newton, A.C., Kang, S., Kato, R.M., Leitges, M., Rawlings, D.J., et al. (2003). A Ras activation pathway dependent on Syk phosphorylation of protein kinase C. Proc Natl Acad USA. 100, 9470-9475. Kenzel, S., Mancuso, G., Malley, R., Teti, G., Golenbock, D.T., and Henneke, P. (2006). c-Jun kinase is a critical signaling molecule in a neonatal model of Group B Streptococcal sepsis. J Immunol. 176, 3181-3188. Keshvara, L.M., Isaacson, C.C., Yankee, T.M., Sarac, R., Harrison, M.L., and Geahlen, R.L. (1998). Syk- and Lyn-dependent phosphorylation of Syk on multiple tyrosines following B cell activation includes a site that negatively regulates signaling. J Immunol. 161, 5276-5283. Kiefer, F., Brumell, J., Al-Alawi, N., Latour, S., Cheng, A., Veillette, A., Grinstein, S., and Pawson, T. (1998). The Syk protein tyrosine kinase is essential for Fcgamma receptor signaling in macrophages and neutrophils, Mol Cell Biol. 18, 4209-4220. Lee, H.-K., Dunzendorfer, S., and Tobias, P.S. (2004). Cytoplasmic domain-mediated dimerizations of Toll-like receptor 4 observed by beta-lactamase enzyme fragment complementation. J Biol Chem. 279, 10564-10574. Li, S., Strelow, A., Fontana, E.J., and Wesche, H. (2002). IRAK-4: A novel member of the IRAK family with the properties of an IRAK-kinase. Proc Natl Acad Sci USA. 99, 5567-5572. Liu, Y., Shepherd, E.G., and Nelin, L.D. (2007). MAPK phosphatases--regulating the immune response. Nat Rev Immunol. 7, 202-212. Loiarro, M., Capolunghi, F., Fanto, N., Gallo, G., Campo, S., Arseni, B., Carsetti, R., Carminati, P., De Santis, R., Ruggiero, V., et al. (2007). Inhibition of MyD88 dimerization and recruitment of IRAK1 and IRAK4 by a novel peptidomimetic compound. J Leukoc Biol. 82, in press. Matsubara, S., Koya, T., Takeda, K., Joetham, A., Miyahara, N., Pine, P., Masuda, E.S., Swasey, C.H., and Gelfand, E.W. (2006). Syk activation in dendritic cells is essential for airway hyperresponsiveness and inflammation. Am J Respir Cell Mol Biol. 34, 426-433. Matsuguchi, T., Masuda, A., Sugimoto, K., Nagai, Y., and Yoshikai, Y. (2003). JNK-interacting protein 3 associates with Toll-like receptor 4 and is involved in LPS-mediated JNK activation. EMBO J. 22, 4455-4464. Meylan, E., Burns, K., Hofmann, K., Blancheteau, V., Martinon, F., Kelliher, M., and Tschopp, J. (2004). RIP1 is an essential mediator of Toll-like receptor 3-induced NF-kappaB activation. Nat Immunol 5, 503-507. Mocsai, A., Humphrey, M.B., Van Ziffle, J.A.G., Hu, Y., Burghardt, A., Spusta, S.C., Majumdar, S., Lanier, L.L., Lowell, C.A., and Nakamura, M.C. (2004). The immunomodulatory adaptor proteins DAP12 and Fc receptor gamma-chain (FcRgamma) regulate development of functional osteoclasts through the Syk tyrosine kinase. Porc Natl Acad Sci USA. 101, 6158-6163. Nakashima, H., Natsugoe, S., Ishigami, S., Okumura, H., Matsumoto, M., Hokita, S., and Aikou, T. (2006). Clinical significance of nuclear expression of spleen tyrosine kinase (Syk) in gastric cancer. Cancer Lett. 236, 89-94. O'Neill, L.A.J., and Bowie, A.G. (2007). The family of five: TIR-domain-containing adaptors in Toll-like receptor signalling. Nat Rev Immunol. 7, 353-364. Oganesyan, G., Saha, S.K., Guo, B., He, J.Q., Shahangian, A., Zarnegar, B., Perry, A., and Cheng, G. (2006). Critical role of TRAF3 in the Toll-like receptor-dependent and -independent antiviral response. Nature 439, 208-211. Oshiumi, H., Matsumoto, M., Funami, K., Akazawa, T., and Seya, T. (2003). TICAM-1, an adaptor molecule that participates in Toll-like receptor 3-mediated interferon beta induction. Nat Immunol. 4, 161-167. Park, J.M., Greten, F.R., Li, Z.-W., and Karin, M. (2002). Macrophage apoptosis by anthrax lethal factor through p38 MAP kinase inhibition. Science. 297, 2048-2051. Poltorak, A., He, X., Smirnova, I., Liu, M.-Y., Huffel, C.V., Du, X., Birdwell, D., Alejos, E., Silva, M., Galanos, C., et al. (1998). Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 282, 2085-2088. Pomerantz, J.L., and Baltimore, D. (1999). NF-kappaB activation by a signaling complex containing TRAF2, TANK and TBK1, a novel IKK-related kinase. EMBO J. 18, 6694-6704. Raeder, E.M.B., Mansfield, P.J., Hinkovska-Galcheva, V., Shayman, J.A., and Boxer, L.A. (1999). Syk activation initiates downstream signaling events during human polymorphonuclear leukocyte phagocytosis. J Immunol. 163, 6785-6793 Sanjuan, M.A., Rao, N., Lai, K.-T.A., Gu, Y., Sun, S., Fuchs, A., Fung-Leung, W.-P., Colonna, M., and Karlsson, L. (2006). CpG-induced tyrosine phosphorylation occurs via a TLR9-independent mechanism and is required for cytokine secretion. J Cell Biol. 172, 1057-1068. Santos-Sierra, S., Golenbock, D.T., and Henneke, P. (2006). Toll-like receptor-dependent discrimination of streptococci. J Endotoxin Res. 12, 307-312. Sato, S., Sanjo, H., Takeda, K., Ninomiya-Tsuji, J., Yamamoto, M., Kawai, T., Matsumoto, K., Takeuchi, O., and Akira, S. (2005). Essential function for the kinase TAK1 in innate and adaptive immune responses. Nat Immunol. 6, 1087-1095. Sato, S., Sugiyama, M., Yamamoto, M., Watanabe, Y., Kawai, T., Takeda, K., and Akira, S. (2003). Toll/IL-1 receptor domain-containing adaptor inducing IFN-beta (TRIF) associates with TNF receptor-associated factor 6 and TANK-binding kinase 1, and activates two distinct transcription factors, NF-kappaB and IFN-regulatory factor-3, in the Toll-like receptor signaling. J Immunol. 171, 4304-4310. Sedlik, C., Orbach, D., Veron, P., Schweighoffer, E., Colucci, F., Gamberale, R., Ioan-Facsinay, A., Verbeek, S., Ricciardi-Castagnoli, P., Bonnerot, C., Tybulewicz, VL., Di, Santo J., and Amigorena, S. (2003). A critical role for Syk protein tyrosine kinase in Fc receptor-mediated antigen presentation and induction of dendritic cell maturation, J Immunol. 170, 846-852. Shaulian, E., and Karin, M. (2002). AP-1 as a regulator of cell life and death. Nat Cell Biol. 4, E131-E136. Shi, Y.H., Tohyama, Y., Kadono, T., He, J.S., Miah, S.M.S., Hazama, R., Tanaka, C., Tohyama, K., and Yamamura, H. (2006). Protein-tyrosine kinase Syk is required for pathogen engulfment in complement-mediated phagocytosis. Blood 107, 4554-4562. Siraganian, R.P., Zhang, J., Suzuki, K., and Sada, K. (2002). Protein tyrosine kinase Syk in mast cell signaling. Mol Immunol. 38, 1229-1233. Songyang, Z., Shoelson, S.E., McGlade, J., Olivier, P., Pawson, T., Bustelo, X.R., Barbacid, M., Sabe, H., Hanafusa, H., and Yi, T. (1994). Specific motifs recognized by the SH2 domains of Csk, 3BP2, fps/fes, GRB-2, HCP, SHC, Syk, and Vav. Mol Cell Biol. 14, 2777-2785. Takada, E., Okahira, S., Sasai, M., Funami, K., Seya, T., and Matsumoto, M. (2007). C-terminal LRRs of human Toll-like receptor 3 control receptor dimerization and signal transmission. Mol Immunol. 44, 3633-3640. Takeda, K., and Akira, S. (2004). TLR signaling pathways. Semin Immunol. 16, 3-9. Takada, Y., and Aggarwal, B.B. (2004). TNF activates Syk protein tyrosine kinase leading to TNF-induced MAPK activation, NF-kappaB activation, and apoptosis. J Immunol. 173, 1066-1077 Takeuchi, O., and Akira, S. (2001). Toll-like receptors; their physiological role and signal transduction system. Int Immunopharmacol. 1, 625-635. Taniguchi, T., Kobayashi, T., Kondo, J., Takahashi, K., Nakamura, H., Suzuki, J., Nagai, K., Yamada, T., Nakamura, S., and Yamamura, H. (1991). Molecular-cloning of a porcine gene syk that encodes a 72-Kda protein-tyrosine kinase showing high susceptibility to proteolysis. J Biol Chem. 266, 15790-15796. Turner, M., Schweighoffer, E., Colucci, F., Di Santo, J.P., and Tybulewicz, V.L. (2000). Tyrosine kinase SYK: essential functions for immunoreceptor signalling. Immunol Today. 21, 148-154. Ulanova, M., Asfaha, S., Stenton, G., Lint, A., Gilbertson, D., Schreiber, A., Befus, D. (2007). Involvement of Syk protein tyrosine kinase in LPS-induced responses in macrophages. J Endotoxin Res. 13, 117-125. Wadleigh, D.J., Reddy, S.T., Kopp, E., Ghosh, S., and Herschman, H.R. (2000). Transcriptional activation of the cyclooxygenase-2 gene in endotoxin-treated RAW 264.7 macrophages. J Biol Chem. 275, 6259-6266. Wang, B.H., Lu, Z.X., and Polya, G.M. (1998). Inhibition of eukaryote serine/threonine-specific protein kinases by piceatannol. Planta Med. 64, 195-199. Werner, S.L., Barken, D., and Hoffmann, A. (2005). Stimulus specificity of gene expression programs determined by temporal control of IKK activity. Science 309, 1857-1861. Xia, Z.-P., and Chen, Z.J. (2005). TRAF2: A double-edged sword? Sci STKE 272, pe7-. Xie, Z.-H., Zhang, J., and Siraganian, R.P. (2000). Positive regulation of c-Jun N-terminal kinase and TNFalpha production but not histamine release by SHP-1 in RBL-2H3 Mast Cells. J Immunol. 164, 1521-1528. Xu, Y., Tao, X., Shen, B., Horng, T., Medzhitov, R., Manley, J.L., and Tong, L. (2000). Structural basis for signal transduction by the Toll/interleukin-1 receptor domains. Nature 408, 111-115. Yamamoto, M., Sato, S., Hemmi, H., Sanjo, H., Uematsu, S., Kaisho, T., Hoshino, K., Takeuchi, O., Kobayashi, M., Fujita, T., Takeda, K., and Akira, S. (2002a). Essential role for TIRAP in activation of the signalling cascade shared by TLR2 and TLR4. Nature 420, 324-329. Yamamoto, M., Sato, S., Mori, K., Hoshino, K., Takeuchi, O., Takeda, K., and Akira, S. (2002b). Cutting Edge: A novel Toll/IL-1 receptor domain-containing adapter that preferentially activates the IFN-beta promoter in the Toll-Like receptor signaling. J Immunol. 169, 6668-6672. Yamamoto, M., Sato, S., Hemmi, H., Hoshino, K., Kaisho, T., Sanjo, H., Takeuchi, O., Sugiyama, M., Okabe, M., Takeda, K., and Akira, S. (2003). Role of adaptor TRIF in the MyD88-independent Toll-like receptor signaling pathway. Science 301, 640-643. Zandi, E., Rothwarf, D.M., Delhase, M., Hayakawa, M., and Karin, M. (1997). The IkappaB kinase complex (IKK) contains two kinase subunits, IKKalpha and IKKbeta, necessary for IkappaB phosphorylation and NF-kappaB activation. Cell 91, 243-252. Zapata, J. M., Lefebvre, S., and Reed, J. C. (2007). Targeting TRAFs for therapeutic intervention. Adv. Exp. Med. Biol. 597, 188-201. Zhang, J., Berenstein, E.H., Evans, R.L., and Siraganian, R.P. (1996). Transfection of Syk protein tyrosine kinase reconstitutes high affinity IgE receptor-mediated degranulation in a Syk-negative variant of rat basophilic leukemia RBL-2H3 cells. J Exp Med. 184, 71-79. Zou, L., Sato, N., and Kone, B.C. (2004). alpha-Melanocyte stimulating hormone protects against H2O2-induced inhibition of wound restitution in IEC-6 cells via a Syk kinase- and NF-kappaB-dependent mechanism. Shock 22, 453-459. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28259 | - |
dc.description.abstract | 在先天免疫系統(innate immune system)中,Toll-like receptors (TLRs)是胞漿內模式識別受體 (pattern recognition receptors, PRRs)家族成員中的一個主要份子,而它的活化反應對於免疫扮演著重要的角色。在免疫系統方面,雖然脾酪氨酸蛋白激酶(Syk)在含免疫酪氨酸受體激活基序 (ITAM)的免疫受體(ITAM-contained immunoreceptor)所引起的訊息傳遞過程中,是一個主要的調控分子,但是Syk在Toll-like receptor (TLR) 訊息傳遞路徑扮演的角色就較不清楚。在此,我們利用研究小鼠的巨噬細胞,在TLR主導的訊息傳導與基因調控的過程,探討Syk在其中扮演角色。不論由小鼠骨髓細胞分化而成的巨噬細胞(BMDM)或是小鼠巨噬細胞株(RAW 264.7 macrophages),TLR的專一辨識配體 [poly(I:C)-TLR3、LPS-TLR4、CpG-TLR9]都可以活化IFNbeta、TNFalpha、MIP2、IL-6、IL-12beta、iNOS及COX-2 mRNA表現,且這些基因表現會受到專一性抑制Syk活性的小分子抑制劑(Syk inhibitor, SykI)及JNK抑制劑所抑制。另外,我們發現TLR3、TLR4、TLR9均有誘發Syk和JNK活化的能力,這是根據增加在Syk (Y519/520)位置自磷酸化、JNK被磷酸化及它們的in vitro激酶能力試驗證據顯示得知。在BMDM及RAW巨噬細胞中,我們發現由TLR所主導而活化的JNK會被SykI及siRNA所抑制,但IKK活化或IkappaBalpha降解過程則不受影響,顯示 Syk為JNK活化的正調控因子。在TLR訊息路徑中,MyD88、TRIF、TRAF3及TRAF6是上游必要的訊息銜接子(adaptor),因此我們使用基因剔除chimera鼠的BMDM研究,Syk與四種分子的訊息傳遞網關係。結果顯示由TLR3/TRIF-, TLR4/MyD88-, TLR4/TRIF-及TLR9/MyD88所主導的JNK活化訊息傳遞路徑中是需要Syk參與在其中。此外TLR3及TLR4活化JNK路徑需要TRAF3及TRAF6,而 TLR9所引起JNK活化只需要TRAF6。為了進一步闡明受體、Syk和TRAFs蛋白之間的關連,我們建構不同區段的質體,包含(deletion of N-terminal SH2 domain, C-terminal SH2 domain, both SH2 domains, or kinase domain) 。在HEK 293T大量表現蛋白的系統下,我們發現Syk能夠與TLR3, TLR4, TLR9, TRAF3及 TRAF6結合,且Syk會利用其kinase domain與TRAF6結合。此外HeNC2巨噬細胞在沒有刺激狀態下,Syk已與TRAF6結合,但當有LPS刺激時,此Syk/TRAF6複合體會被徵召到TLR4受體。總括而言,目前結果顯示在TLR主導JNK活化訊息路徑中,Syk是一個必要分子,對於IKK活化路徑則非必要。這一個訊息任務依靠著Syk與TRAF6蛋白互相結合,且當LPS刺激情況下,Syk會被徵召到TLR4經過TLR4-MyD88或 TRIF-TRAF6複合體形成進而調控下游JNK活化的訊息傳遞。 | zh_TW |
dc.description.abstract | Toll-like receptor (TLR) is a major family of pattern recognition receptors (PRRs) and plays a crucial role in innate immune system. Even though non-receptor spleen tyrosine kinase (Syk) is a key signaling molecule of ITAM-contained immunoreceptors, its role in TLRs signaling is not clearly understood. Herein, we investigated the role of Syk in TLR-mediated signaling and gene regulation in murine macrophages. In both mouse bone marrow-derived macrophages (BMDM) and murine RAW 264.7 macrophages, poly (I:C), LPS and CpG, which are specific ligands for TLR3, TLR4 and TLR9 respectively, can increase the mRNA levels of several pro-inflammatory cytokines and mediators, including IFNbeta, TNFalpha, MIP2, IL-6 and IL-12beta, iNOS and COX-2. These transcriptional gene upregulation caused by TLR activation were inhibited by specific Syk inhibitor (SykI) and JNK inhibitor (SP600125). Accordingly we found the abilities of TLR3, TLR4 and TLR9 to induce Syk and JNK activation, as evidenced by the increase of Syk auto-phosphorylation on Y519/Y520, JNK phosphorylation as well as their in vitro kinase activities. We also found that TLRs-mediated activation of JNK, but not IKK or IkappaBalpha degradation in BMDM and RAW 264.7 macrophages was blocked by SykI and Syk siRNA, suggesting that Syk is a positive regulator of JNK. Since MyD88, TRIF, TRAF3 and TRAF6 are crucial upstream molecules for TLR signaling, we thus used BMDM isolated from genetic knockout chimera mice to study the coupling between Syk and these adaptors. Results revealed that Syk is involved in TLR3/TRIF-, TLR4/MyD88-, TLR4/TRIF- and TLR9/MyD88-mediated JNK signaling. In addition, both TRAF3 and TRAF6 contribute to JNK activation elicited by TLR3 and TLR4, while only TRAF6 is required for JNK activation by TLR9. To further dissect the protein interactions among receptors, Syk and TRAFs, we constructed Syk plasmids with different domains (deletion of N-terminal SH2 domain, C-terminal SH2 domain, both SH2 domains, or kinase domain. We found that in expression cell system Syk is able to associate with TLR3, TLR4, TLR9, TRAF3 and TRAF6. Moreover, kinase domain of Syk is required for its association with TRAF6. In resting HeNC2 macrophages, Syk can associate with TRAF6, and upon LPS stimulation this Syk/TRAF6 complex is recruited to TLR4. All together, our data highlight the role of Syk in coupling TLR signaling to JNK but not to IKK pathway. This signaling role relies on the interaction of Syk to TRAF6, and upon LPS stimulation, Syk can be recruited to TLR4 through the TLR4-adaptors (MyD88 or TRIF)-TRAF6 complex formation, and then regulates downstream JNK signal. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T00:03:48Z (GMT). No. of bitstreams: 1 ntu-96-R94443022-1.pdf: 1899332 bytes, checksum: 45590e3754a74fdab5cbd2ad88e057a5 (MD5) Previous issue date: 2007 | en |
dc.description.tableofcontents | Abbreviations...…………………………………………………………………….…..1
Abstract…………………………………………………………………………..……..4 中文摘要 ..…………………..……………………………………………………..…..6 Introduction……………………………………………………………………….……8 Materials and Methods…………..…………………………………………………...26 Results………………………………………………………………………………... .38 Discussion……...………………………………………………………………………48 Figures….…………...…………………………………………………………………55 Appendix…………………………………………………………………………...…..70 References……………………………………………………...………..……………..74 | |
dc.language.iso | en | |
dc.title | 探討巨噬細胞Spleen tyrosine kinase在Toll-Like Receptors的訊息傳遞及誘發的發炎基因表現中所扮演的角色 | zh_TW |
dc.title | Roles of spleen tyrosine kinase in TLRs-mediated signal transduction and induced inflammatory gene expression in macrophages | en |
dc.type | Thesis | |
dc.date.schoolyear | 95-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 朱清良(Ching-Liang Chu),顏茂雄(Mao-Hsiung Yen),楊春茂(Chuen-Mao Yang),蘇銘嘉(Ming-Jai Su) | |
dc.subject.keyword | 先天免疫反應,發炎反應,脾酪氨酸激酶,促發炎細胞素, | zh_TW |
dc.subject.keyword | TLR,Syk,innate immune,TRAF,pro-inflammatory cytokines,PRR, | en |
dc.relation.page | 87 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2007-07-31 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 藥理學研究所 | zh_TW |
Appears in Collections: | 藥理學科所 |
Files in This Item:
File | Size | Format | |
---|---|---|---|
ntu-96-1.pdf Restricted Access | 1.85 MB | Adobe PDF |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.