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???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
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dc.contributor.advisor | 蔡錦華 | |
dc.contributor.author | Wen-Yi Luo | en |
dc.contributor.author | 羅文宜 | zh_TW |
dc.date.accessioned | 2021-06-13T03:37:47Z | - |
dc.date.available | 2006-08-04 | |
dc.date.copyright | 2006-08-04 | |
dc.date.issued | 2006 | |
dc.date.submitted | 2006-07-26 | |
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Serum cytokine levels in i | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32228 | - |
dc.description.abstract | 在被EB病毒(Epstein-Barr virus)感染的B細胞中,EB病毒利用抑制細胞凋亡和促使細胞持續生長兩個方式來建立類淋巴母細胞株(Lymphoblastoid cell line)。本論文的目的在於研究受到EB病毒調控,並參與在抑制細胞凋亡或促使細胞持續生長的細胞蛋白質。利用核酸微陣列晶片(microarray)來分析被EB病毒感染的B細胞中凋亡抑制蛋白質家族(inhibitor of apoptosis protein famiy)的變化,發現survivin轉錄因子(transcript)有顯著地增加。另外,在EB病毒感染血液單核細胞(mononuclear cells)的過程中,利用同步定量聚合酶連鎖反應(real-time quantitative PCR)來偵測四個凋亡抑制蛋白質表現,包括HIAP-1, HIAP-2, XIAP和survivin。實驗結果發現survivin轉錄因子會隨著感染時間的延長而增加表現量。同時,被EB病毒感染七天後的血液單核細胞中,也可以偵測到survivin蛋白質表現。更進一步地發現survivin轉錄因子增加的現象是藉由EB病毒核抗原蛋白質第二型(EBNA2)來調控。根據以上觀察,認為EB病毒可以利用EBNA2來調控survivin的表現,藉此促進細胞存活。
在促進細胞生長過程中,細胞激素(cytokine)扮演著一個重要的因子。在本論文中,藉由人類細胞激素抗體微陣列(human cytokine antibody array),觀察受EB病毒感染血液單核細胞的培養液中,細胞激素的變化。根據實驗的結果,觀察到在受EB病毒感染血液單核細胞的培養液中,GRO, IL-6, IL-10, IL-13, MCP-2和RANTES的表現量有顯著的增加。本論文著重於參與B細胞增生的細胞激素,而以IL-13作為目標蛋白質。在EB病毒感染血液單核細胞的二十八天周期中,利用同步定量聚合酶連鎖反應和酵素連結免疫吸附法(ELISA),發現IL-13轉錄因子和蛋白質的表現在病毒感染之後都有上升的現象。然而,IL-13轉錄因子和蛋白質的表現在感染末期則有下降的趨勢。另外,在EB病毒感染的血液單核細胞培養液中加入IL-13中和抗體,觀察到被感染細胞的增生能力降低。综合以上結果,推測EB病毒感染細胞後,可以利用分泌IL-13來幫助B細胞的增生。 | zh_TW |
dc.description.abstract | Anti-apoptosis and unlimiting proliferation are curial steps for Epstein-Barr virus (EBV) immortalization. In this thesis, the cellular proteins induced by EBV which may participate in prevention of cell death or stimulation of cell growth were investigated. The expression of IAPs, an anti-apoptotic protein family, was examined in EBV-infected B cells using microarray analysis. Survivin was the obvious target which was increase after virus infection. In addition, the kinetics of IAPs, including HIAP-1, HIAP-2, XIAP and survivin were further investigated in EBV-infected peripheral blood mononuclear cells (PBMCs) by real-time quantitative PCR (Q-PCR). Survivin transcript was up-regulated after EBV infection in a time-dependent manner. Moreover, survivin protein was also detected in EBV-infected PBMCs at day 7 post infection. Furthermore, EBV nuclear antigen, EBNA2, induced expression of survivin transcript in PBMCs using transient transfection. These data suggest that EBV may utilize EBNA2 to regulate survivin expression in order to facilitate the cell survival.
Cell proliferation is stimulating by cytokine expression. In this thesis, the differential expression of cytokine in the supernatant of PBMCs upon EBV infection was analyzed using human cytokine antibody array method. The expression of GRO, IL-6, IL-10, IL-13, MCP-2 and RANTES were increased after virus infection. IL-13, which is able to stimulate the growth of B cells, was the target we were interested in. The kinetics of IL-13 transcript and protein were determined in EBV-infected PBMCs by Q-PCR and ELISA, respectively. In twenty-eight days of infection period, IL-13 was increased at the middle of period but decreased at the end in both transcriptional and translational levels. Moreover, the proliferation of EBV-infected PBMCs was declined after treatment of IL-13 neutralizing antibody. It suggests that IL-13 may involve in the growth of EBV-immortalized cells. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T03:37:47Z (GMT). No. of bitstreams: 1 ntu-95-R93445116-1.pdf: 1233896 bytes, checksum: c86e7af17689acef6d8d42b23ec9a37f (MD5) Previous issue date: 2006 | en |
dc.description.tableofcontents | Introduction………………........………………………………………………………1
EBV……………………………….....……………………………………………………1 Historical background…………......……………………………………………………1 Establishment of EBV latent infection in B cells………………………………............1 Characteristics of EBV-immortalized B cells…………………………………….........2 EBV and anti-apoptosis……………………………………………………………........3 Apoptotic death pathway………………………………………………………….........3 Bcl-2 family………………………………………………………………………….....3 Regulation of Bcl-2 family by EBV………………………………………………........4 Inhibition of apoptosis protein family………………………………………………......5 Historical background………………………………………………………………......5 Structure…………………………………………………………………………….......5 Inhibition mechanism……………………………………………………………........5 Survivin………...……………………………………………………………………........6 Characteristics and tissue distribution……………………………………………….....6 Structure…………………………………………………………………………….......6 Regulation ………………………………………………………………………….......6 Function..…………………………………………………………………………….....7 IL-13…………………………………………………………………………………........8 Tissue distribution…………………………………………………………………........8 Receptors………………………………………………………………………..............8 Signaling pathway…………………………………………………………………........9 Function…………………………………………………………………………...........9 Aims of the study………………..…………………......………………………………..10 Materials and Methods……………………….........……………....………………11 Cell culture…………………………………………………………………….........…11 Virus preparation, titration and infection………………………………………...........11 PBMCs isolation ………………………………………………………………......….12 RNA extraction ………………………………………………………………….....…12 Reverse transcription…………………………………………………………….....…12 Q-PCR…………......................................................…………………………………..13 Western blotting assay………………………….....…………………………………..13 Plasmid preparation…………………………….....…………………………………..14 DNA electroporation……………………………......…………………………………14 B cell purification and microarray experiment…......…………………………………15 Human cytokine antibody array…………..……......………………………………….15 ELISA…..................................................……………………..................……………15 [3H]-thymidine incorporation assay………………...……......………………………..16 Results…………….........………………………………………………………………17 Expression profiles of IAP gene family in EBV infected B cells……........…………..17 Expression of survivin transcript and protein in kinetics upon EBV infection….........17 EBNA2 involved in the regulation of survivin…………………………………..........18 Expression profiles of cytokine and chemokine in EBV infected PBMCs…….......…18 Expression of IL-13 transcript and protein in kinetics upon EBV infection….......…..19 IL-13 promoted proliferation of EBV-infected PBMCs……………………….....…...20 Discussion………………….......……………………………………………………...21 Regulation of survivin by EBV……........……………………………………………..21 Regulation of IL-13 by EBV………........……………………………………………..23 Tables and Figures………………………......………………...……………………25 References……………………………………......……………………………………39 | |
dc.language.iso | en | |
dc.title | 探討凋亡抑制蛋白質及細胞激素在EB病毒感染過程之變化 | zh_TW |
dc.title | To investigate the expression profile of inhibitor of apoptosis proteins and cytokines during Epstein-Barr virus infection | en |
dc.type | Thesis | |
dc.date.schoolyear | 94-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 呂春敏,李建國 | |
dc.subject.keyword | 凋亡抑制蛋白質,細胞激素,EB病毒,感染, | zh_TW |
dc.subject.keyword | EBV,survivin,IL-13,apoptosis,proliferation, | en |
dc.relation.page | 49 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2006-07-27 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 微生物學研究所 | zh_TW |
Appears in Collections: | 微生物學科所 |
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