(3.230.143.40) 您好!臺灣時間:2021/04/19 04:43
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果

詳目顯示:::

我願授權國圖
: 
twitterline
研究生:蔡依螢
研究生(外文):Yi-Ying Tsai
論文名稱:克雷白氏肺炎桿菌感染肝細胞HepG2在TLR與MAPK間訊息分子研究
論文名稱(外文):Studies of signaling molecules between TLR and MAPK in HepG2 cells infected with Klebsiella pneumoniae
指導教授:謝絹珠
指導教授(外文):June-Hsieh Wu
學位類別:碩士
校院名稱:長庚大學
系所名稱:基礎醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2006
畢業學年度:95
語文別:中文
論文頁數:68
中文關鍵詞:克雷白氏肺炎桿菌莢膜多醣體化膿性肝膿瘍
外文關鍵詞:Klebsiella pneumoniaeMAPKToll-like receptorsHepG2TLRsliver abscessp44/42p38
相關次數:
  • 被引用被引用:0
  • 點閱點閱:219
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
在台灣,克雷白氏肺炎桿菌 (klebsiella pneunomina, KP) 是化膿性肝膿瘍 (pyogenic liver abscess) 的主要致病菌,國外少見此種現象。實驗室的興趣在於釐清KP與肝細胞的關係和KP在肝細胞中引起的訊息傳遞現象。之前實驗室利用HepG2作為感染系統,發現活菌KP可引起細胞當中的p44/42、p38傳遞路徑之活化和TLR2/TLR4高度的表現;在此實驗中我們由KP分離的莢膜多醣體 (capsule polysaccharide) 單獨存在發現可活化p44/42、p38,但只刺激TLR4表現。在抗體抑制性試驗 (感染後30分鐘) 的結果當中,發現抑制TLR2及TLR4後,HepG2細胞之p44/42 MAPK及p38 kinase路徑之活化可被少量的抑制,但以純化之KP LPS來與細胞作用時,TLR4抗體可明顯的顯示p44/42及p38路徑被抑制。猜想在活菌作用時,莢膜多醣體的立體結構可能部份擋住位於克雷白氏肺炎桿菌上TLR2和TLR4的配體 (ligand),另外或許也可能有其他受體在全菌感染時參與下游p44/42和p38的磷酸化,致使反應不明顯。在免疫沉澱試驗中,以KP LPS與HepG2作用(感染後30分中),可觀察到MyD88及TRAF6相互作用,但未能觀察到MyD88與TAK1的交互作用,此部分需進一步的研究。
Severe liver abscess caused by Klebsiella pneumoniae (KP) has been observed recently in Taiwan. How the bacteria cause liver abscess is not clear. Our laboratory previously used human hepatocellular carcinoma cell line, HepG2, as a model system to study the signal transduction during infection and found that infecting HepG2 cell with whole KP bacteria caused upregulation of TLR2 and TLR4 protein expression and activation of p38 kinase and p44/42 MAPK pathways. In this experiment, we found that bacterial capsule polysaccharide purified from KP can activated p44/42, p38 and TLR4 but not TLR2. We did not observe the downstream molecules, MyD88, IRAK1, TRAF6, TAK1 interacting with TLR2 or TLR4 in HepG2 cells infected with live whole bacteria by immunoprecipitation experiment. However, in HepG2 cells treated with KP LPS, the interaction of Myd88 and TRAF6 could be observed. Inhibition assay with TLR2 or TLR4 antibodies indicated that both p38 kinase and p44/42 MAPK activations were inhibited slightly at antibody concentration of 20μg/ml, however, such inhibition can be observed clearly when the whole bacteria infection was substituted with LPS form KP. It seems that capsule polysaccharide may form steric hinderance to prevent bacterial LPS from activation downstream molecules. The association of other molecules, such as IRAK1, TAK1, MyD88 and TRAF6 with TLR2 or TLR4 needs further investigation.
中文摘要
英文摘要
目錄
緒論 1
ㄧ、克雷白氏肺炎桿菌的介紹 1
二、台灣地區的克雷白是肺炎桿菌特有的肝膿 2
三、克雷白氏肺炎桿菌的致病因子 3
四、由細菌感染引發的訊息傳遞 8
甲、Toll-like receptors 8
TLR4 10
TLR2 10
TLRs所引發的訊息傳遞 11
乙、MAPK kinase 14
丙、NF-κB 16
研究目的 21
實驗材料與方法 22
實驗結果 27
討論 54
結論 61
參考文獻 62
Akira, S. (2003) Mammalian Toll-like receptors. Curr Opin Immunol. 15: 5-11.
Akira, S. and Sato, S. (2003) Toll-like receptors and their signaling mechanisms. Scand J Infect Dis. 35: 555-562.
Akira, S. and Takeda, K. (2004) Toll-like receptor signalling. Nat Rev Immunol. 4: 499-511.
Akira, S., Takeda, K. and Kaisho, T. (2001) Toll-like receptors: critical proteins linking innate and acquired immunity. Nat Immunol. 2: 675-680.
Batshon, B.A., Baer, H. and Shaffer, M.F. (1963) Immunologic paralysis produced in mice by Klebsiella pneumoniae type 2 polysaccharide. J Immunol. 90: 121-126.
Bochud, P.Y. and Calandra, T. (2003) Pathogenesis of sepsis: new concepts and implications for future treatment. Bmj. 326: 262-266.
Bonizzi, G. and Karin, M. (2004) The two NF-kappaB activation pathways and their role in innate and adaptive immunity. Trends Immunol. 25: 280-288.
Chan, K.S., Chen, C.M., Cheng, K.C., Hou, C.C., Lin, H.J. and Yu, W.L. (2005) Pyogenic liver abscess: a retrospective analysis of 107 patients during a 3-year period. Jpn J Infect Dis. 58: 366-368.
Cheng, H.P., Chang, F.Y., Fung, C.P. and Siu, L.K. (2002) Klebsiella pneumoniae liver abscess in Taiwan is not caused by a clonal spread strain. J Microbiol Immunol Infect. 35: 85-88.
Cryz, S.J., Jr., Furer, E. and Germanier, R. (1986) Immunization against fatal experimental Klebsiella pneumoniae pneumonia. Infect Immun. 54: 403-407.
Darveau, R.P., Pham, T.T., Lemley, K., Reife, R.A., Bainbridge, B.W., Coats, S.R., et al (2004) Porphyromonas gingivalis lipopolysaccharide contains multiple lipid A species that functionally interact with both toll-like receptors 2 and 4. Infect Immun. 72: 5041-5051.
Denkers, E.Y., Butcher, B.A., Del Rio, L. and Kim, L. (2004) Manipulation of mitogen-activated protein kinase/nuclear factor-kappaB-signaling cascades during intracellular Toxoplasma gondii infection. Immunol Rev. 201: 191-205.
Fader, R.C. and Davis, C.P. (1980) Effect of piliation on Klebsiella pneumoniae infection in rat bladders. Infect Immun. 30: 554-561.
Fader, R.C. and Davis, C.P. (1982) Klebsiella pneumoniae-induced experimental pyelitis: the effect of piliation on infectivity. J Urol. 128: 197-201.
Fan, J., Frey, R.S. and Malik, A.B. (2003) TLR4 signaling induces TLR2 expression in endothelial cells via neutrophil NADPH oxidase. J Clin Invest. 112: 1234-1243.
Faure, E., Thomas, L., Xu, H., Medvedev, A., Equils, O. and Arditi, M. (2001) Bacterial lipopolysaccharide and IFN-gamma induce Toll-like receptor 2 and Toll-like receptor 4 expression in human endothelial cells: role of NF-kappa B activation. J Immunol. 166: 2018-2024.
Frantz, S., Kobzik, L., Kim, Y.D., Fukazawa, R., Medzhitov, R., Lee, R.T. and Kelly, R.A. (1999) Toll4 (TLR4) expression in cardiac myocytes in normal and failing myocardium. J Clin Invest. 104: 271-280.
Fung, C.P., Hu, B.S., Chang, F.Y., Lee, S.C., Kuo, B.I., Ho, M., et al (2000) A 5-year study of the seroepidemiology of Klebsiella pneumoniae: high prevalence of capsular serotype K1 in Taiwan and implication for vaccine efficacy. J Infect Dis. 181: 2075-2079.
Fung, C.P., Chang, F.Y., Lee, S.C., Hu, B.S., Kuo, B.I., Liu, C.Y., et al (2002) A global emerging disease of Klebsiella pneumoniae liver abscess: is serotype K1 an important factor for complicated endophthalmitis? Gut. 50: 420-424.
Grube, B.J., Cochane, C.G., Ye, R.D., Green, C.E., McPhail, M.E., Ulevitch, R.J. and Tobias, P.S. (1994) Lipopolysaccharide binding protein expression in primary human hepatocytes and HepG2 hepatoma cells. J Biol Chem. 269: 8477-8482.
Hadley, J.S., Wang, J.E., Foster, S.J., Thiemermann, C. and Hinds, C.J. (2005) Peptidoglycan of Staphylococcus aureus upregulates monocyte expression of CD14, Toll-like receptor 2 (TLR2), and TLR4 in human blood: possible implications for priming of lipopolysaccharide signaling. Infect Immun. 73: 7613-7619.
Held, T.K., Trautmann, M., Mielke, M.E., Neudeck, H., Cryz, S.J., Jr. and Cross, A.S. (1992) Monoclonal antibody against Klebsiella capsular polysaccharide reduces severity and hematogenic spread of experimental Klebsiella pneumoniae pneumonia. Infect Immun. 60: 1771-1778.
Jeyaseelan, S., Young, S.K., Yamamoto, M., Arndt, P.G., Akira, S., Kolls, J.K. and Worthen, G.S. (2006) Toll/IL-1R domain-containing adaptor protein (TIRAP) is a critical mediator of antibacterial defense in the lung against Klebsiella pneumoniae but not Pseudomonas aeruginosa. J Immunol. 177: 538-547.
Ko, W.C., Paterson, D.L., Sagnimeni, A.J., Hansen, D.S., Von Gottberg, A., Mohapatra, S., et al (2002) Community-acquired Klebsiella pneumoniae bacteremia: global differences in clinical patterns. Emerg Infect Dis. 8: 160-166.
Lederman, E.R. and Crum, N.F. (2005) Pyogenic liver abscess with a focus on Klebsiella pneumoniae as a primary pathogen: an emerging disease with unique clinical characteristics. Am J Gastroenterol. 100: 322-331.
Lin, J.C., Chang, F.Y., Fung, C.P., Xu, J.Z., Cheng, H.P., Wang, J.J., et al (2004) High prevalence of phagocytic-resistant capsular serotypes of Klebsiella pneumoniae in liver abscess. Microbes Infect. 6: 1191-1198.
Liu, S., Gallo, D.J., Green, A.M., Williams, D.L., Gong, X., Shapiro, R.A., et al (2002) Role of toll-like receptors in changes in gene expression and NF-kappa B activation in mouse hepatocytes stimulated with lipopolysaccharide. Infect Immun. 70: 3433-3442.
Liu, Y., Wang, Y., Yamakuchi, M., Isowaki, S., Nagata, E., Kanmura, Y., et al (2001) Upregulation of toll-like receptor 2 gene expression in macrophage response to peptidoglycan and high concentration of lipopolysaccharide is involved in NF-kappa b activation. Infect Immun. 69: 2788-2796.
Lodish, H.B., Arnold; Zipursky, S. Lawrence; Matsudaira, Paul; Baltimore, David; Darnell, James E. (2004) Molecular cell biology: Freeman.
McDonald, M.I., Corey, G.R., Gallis, H.A. and Durack, D.T. (1984) Single and multiple pyogenic liver abscesses. Natural history, diagnosis and treatment, with emphasis on percutaneous drainage. Medicine (Baltimore). 63: 291-302.
Merino, S., Camprubi, S., Alberti, S., Benedi, V.J. and Tomas, J.M. (1992) Mechanisms of Klebsiella pneumoniae resistance to complement-mediated killing. Infect Immun. 60: 2529-2535.
Muzio, M., Bosisio, D., Polentarutti, N., D'Amico, G., Stoppacciaro, A., Mancinelli, R., et al (2000) Differential expression and regulation of toll-like receptors (TLR) in human leukocytes: selective expression of TLR3 in dendritic cells. J Immunol. 164: 5998-6004.
Nanbo, A., Nishimura, H., Muta, T. and Nagasawa, S. (1999) Lipopolysaccharide stimulates HepG2 human hepatoma cells in the presence of lipopolysaccharide-binding protein via CD14. Eur J Biochem. 260: 183-191.
Palsson-McDermott, E.M. and O'Neill, L.A. (2004) Signal transduction by the lipopolysaccharide receptor, Toll-like receptor-4. Immunology. 113: 153-162.
Podschun, R. and Ullmann, U. (1998) Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev. 11: 589-603.
Regueiro, V., Campos, M.A., Pons, J., Alberti, S. and Bengoechea, J.A. (2006) The uptake of a Klebsiella pneumoniae capsule polysaccharide mutant triggers an inflammatory response by human airway epithelial cells. Microbiology. 152: 555-566.
Rubin (1994) pathology.
Sahly, H., Podschun, R., Oelschlaeger, T.A., Greiwe, M., Parolis, H., Hasty, D., et al (2000) Capsule impedes adhesion to and invasion of epithelial cells by Klebsiella pneumoniae. Infect Immun. 68: 6744-6749.
Saitoh, S., Akashi, S., Yamada, T., Tanimura, N., Kobayashi, M., Konno, K., et al (2004) Lipid A antagonist, lipid IVa, is distinct from lipid A in interaction with Toll-like receptor 4 (TLR4)-MD-2 and ligand-induced TLR4 oligomerization. Int Immunol. 16: 961-969.
Schaeffer, H.J. and Weber, M.J. (1999) Mitogen-activated protein kinases: specific messages from ubiquitous messengers. Mol Cell Biol. 19: 2435-2444.
Silvain, C. (2001) [Liver abscess]. Rev Prat. 51: 2069-2074.
Simoons-Smit, A.M., Verweij-van Vught, A.M. and MacLaren, D.M. (1986) The role of K antigens as virulence factors in Klebsiella. J Med Microbiol. 21: 133-137.
Takeda, K. and Akira, S. (2005) Toll-like receptors in innate immunity. Int Immunol. 17: 1-14.
Underhill, D.M. (2004) Toll-like receptors and microbes take aim at each other. Curr Opin Immunol. 16: 483-487.
Werts, C., Tapping, R.I., Mathison, J.C., Chuang, T.H., Kravchenko, V., Saint Girons, I., et al (2001) Leptospiral lipopolysaccharide activates cells through a TLR2-dependent mechanism. Nat Immunol. 2: 346-352.
Wu, J.H., Hong, L.C., Tsai, Y.Y., Chen, H.W., Chen, W.X. and Wu, T.S. (2006) Mitogen-activated protein kinase (MAPK) signalling pathways in HepG2 cells infected with a virulent strain of Klebsiella pneumoniae. Cell Microbiol. 8: 1467-1474.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關期刊
 
系統版面圖檔 系統版面圖檔