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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 朱清良(Ching-Liang Chu) | |
dc.contributor.author | Hsueh-Han Lu | en |
dc.contributor.author | 呂學翰 | zh_TW |
dc.date.accessioned | 2021-05-15T17:52:18Z | - |
dc.date.available | 2014-10-09 | |
dc.date.available | 2021-05-15T17:52:18Z | - |
dc.date.copyright | 2014-10-09 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-08-12 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/5128 | - |
dc.description.abstract | 半乳糖凝集素 (Galectins) 是一種動物性凝集素,可以專一地辨識β-半乳糖 (β-galactosides)。 在免疫系統中,galctin-1 和 -3已經被報導可以調控各種不同的免疫細胞的免疫反應,然而galectin-1和 -3對於漿狀樹突細胞 (plasmacytoid dendritic cell, pDC) 之生長與功能的影響尚未釐清。 pDC在受到病毒感染時,能夠分泌大量的第一型干擾素 (type I Interferon, IFN-I) 並引發各種抗病毒的活性。因此,我們除了研究galectin-1和 -3對於pDC的生長影響外,也用類鐸受體7/9的配體,R848或CpG刺激pDC,以研究pDC產生IFN-I的能力是否會受到galectin-1和 -3的影響。在初步的實驗結果中,我們發現不論是活體外 (in vitro) 或是活體內 (in vivo) 的試驗,皆發現galectin-1和 -3對於pDC的生長沒有顯著的影響。此外,半乳糖凝集素-3基因剔除 (lgals3-/-) pDC在CpG刺激後,其第一型干擾素IFN-I (ifnα, ifnα4, ifnβ)和干擾素調控因子7 (Interferon regulatory factor 7, IRF7) 的基因表現量的增加顯著高於野生型 (wild type, WT) pDC。而且,lgals3-/- pDC在R848或CpG刺激後所產生的IFN-I也比WT pDC來的高。與lgals3-/- pDC相反的是lgals1-/- pDC,在R848或CpG刺激後,其IFN-I 基因表現量的增加顯著低於WT pDC,另外產生的IFN-I 也較WT pDC來的低。因此,我們推測galectin-3 在pDC中可能是扮演了負向調控其產生IFN-I 功能的角色,而galectin-1 則是扮演了正向調控的角色。然而,galectin-1和 -3是經由何種機轉調控IFN-I 的產生,仍然需要進一步的研究。 | zh_TW |
dc.description.abstract | Galectins are animal lectins that can bind to β-galactosides. In the immune system, galectin-1 and galectin-3 have been shown to modulate immune responses in various types of immune cell. However, the role of galectin-1 and galectin-3 in the development and the function of plasmacytoid dendritic cell (pDC) has not yet been studied. pDCs produce type-1-interferon (IFN-I) in response to viral infection and then enhance antiviral activities. Thus, we examined the effect of galectin-1 and galectin-3 on pDC development and IFN-I production by pDCs upon the stimulation of R848 or CpG through TLR7/9. In preliminary results, we show that galectin-1 and galectin-3 didn’t influence the development of pDC in vitro or in vivo. In addition, we found that CpG or R848-induced mRNA levels of IFN-I (ifnα, ifnα4, ifnβ) and interferon regulatory factor (IRF7) were significantly higher in gal-3-/- Flt3L-BMDCs and in the contrast, significantly lower in gal-1-/- Flt3L-BMDCs when compared to that in WT Flt3L-BMDCs. Moreover, at the protein level, the production of IFNα by gal-3-/- Flt3L-BMDCs stimulated with CpG or R848 is higher than WT Flt3L-BMDCs. And, the production of IFNα by gal-3-/- Flt3L-BMDCs is lower than WT Flt3L-BMDCs. Our data suggested that galectin-3 may negatively regulate the function of pDCs, on the other hand that galectin-1 may positively regulate the function of pDC. However, the mechanism of how galectin-3 and galectin-1 involved in IFN-I production need to be further explored. | en |
dc.description.provenance | Made available in DSpace on 2021-05-15T17:52:18Z (GMT). No. of bitstreams: 1 ntu-103-R01449010-1.pdf: 3926646 bytes, checksum: 9f681fd554d256546e8c8441ccb4e2e9 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 誌謝 i
摘要 ii Abstract iii Abbreviations iv Table of contents v List of figures viii Chapter I Introduction 1 1.1 The role of galectin-1 and galectin-3 in DC 2 1.2 pDC 3 1.3 Development of pDC 4 1.4 The main function of pDC 5 1.5 TLR7/9 dependent signaling pathway 6 1.6 Specific aims 7 Chapter II Materials and Methods 11 2.1 Mice 12 2.2 In vitro pDC culture and DC subsets analysis 12 2.3 In vivo expansion of pDCs 13 2.4 Splenic DC isolation and DC subsets analysis 13 2.5 Functional assays of pDCs 13 2.6 qPCR 14 2.7 Bioassay for IFN-I 14 2.8 Assessment of cytokine production by ELISA 15 2.9 Flow cytometry 15 2.10 PMDC05 cell line 15 2.11 Statistical analysis 15 Chapter III Results 16 3.1 Establishing in vitro pDC culture system 17 3.2 Expansion of pDC in vivo 17 3.3 Positive control of IFNα intracellular staining 17 3.4 Galectins expression in pDC 18 3.5 Galectin expression profile in human pDC cell line, PMDC05 18 3.6 Galectin-3 doesn’t get involved in mouse pDC development 19 3.7 Gal-3-/- Flt3L-BMDCs exhibit augmented IFN-I and IL-12 production in response to CpG or R848 19 3.8 Sorted Flt3L-BMpDC from Gal-3-/- mice had higher IFN-I production after stimulation with CpG or R848 21 3.9 Sorted splenic pDC from Gal-3-/- mice had higher IFN-I production after stimulation with R848 22 3.10 Galectin-1 doesn’t get involved in mouse pDC development 22 3.11 Gal-1-/- Flt3L-BMDCs exhibit an impaired IFN-I response after CpG or R848 stimulation 23 Chapter IV Discussion 24 4.1 Distinct roles of galectin-1 and -3 in regulating the production of IFN-I by pDC 25 4.2 Galectin expression profile of human pDC cell line, PMDC05 27 4.3 Autophagosome involved in transferring the nucleic-acid to TLR 27 4.4 Functionless sorted splenic pDC 28 4.5 phosphorylation of galectin 28 Figures 30 References 62 | |
dc.language.iso | en | |
dc.title | 半乳糖凝集素-1和半乳糖凝集素-3在調控漿狀樹突細胞發育及功能之角色 | zh_TW |
dc.title | The Role of Galectin-1 and Galectin-3 in Development and Function of Plasmacytoid Dendritic Cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 劉扶東(Fu-Tong Liu),李建國(Chien-Kuo Lee) | |
dc.subject.keyword | 漿狀樹突細胞,半乳糖凝集素-1,半乳糖凝集素-3,第一型干擾素,類鐸受體7/9, | zh_TW |
dc.subject.keyword | plasmacytoid dendritic cell,galectin-1,galectin-3,type-I-IFN,TLR7/9, | en |
dc.relation.page | 74 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2014-08-12 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 免疫學研究所 | zh_TW |
顯示於系所單位: | 免疫學研究所 |
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ntu-103-1.pdf | 3.83 MB | Adobe PDF | 檢視/開啟 |
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