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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 江伯倫(Bor-luen Chiang) | |
dc.contributor.author | Yi-chen Tsai | en |
dc.contributor.author | 蔡宜臻 | zh_TW |
dc.date.accessioned | 2021-06-15T11:43:26Z | - |
dc.date.available | 2021-08-26 | |
dc.date.copyright | 2016-08-26 | |
dc.date.issued | 2016 | |
dc.date.submitted | 2016-08-15 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49710 | - |
dc.description.abstract | Treg-of-B細胞是一群具有抑制能力的調節性T細胞,他們和其他調節性T細胞不同,不同於第一型調節性T細胞不同的地方在於Treg-of-B是一群CD25陽性的細胞,而第一型調節性T細胞則是CD25陰性,在轉錄因子Foxp3的表現上,Treg-of-B並不表現Foxp3,這個部分與自然調節性T細胞亦不相同,而目前為止,關於Treg-of-B的研究有限,所以我們希望能夠更加了解Treg-of-B在免疫抑制上的功能。半乳糖凝集素是一群會和-galactoside結合的蛋白,目前有愈來愈多文獻指出,他們和先天免疫與後天免疫之間都有相關性,例如半乳糖凝集素-2能夠造成CD8陽性T細胞的細胞凋亡來達到抑制免疫反應,但在先天免疫上,半乳糖凝集素卻扮演一個促進發炎的角色。
而從我們的實驗中發現,Treg-of-B會高度表現半乳糖凝集素-2的基因,而自然調節性T細胞的表現量則和naïve T cell相差不多,接著我們進一步研究半乳糖凝集素-2如何去抑制T細胞增生,於是藉由阻斷naïve T cell表面上半乳糖凝集素-2的可能結合受體CD29來看是否會影響半乳糖凝集素-2的抑制能力,而實驗結果發現,實驗組和對照組並無差異,我們推測半乳糖凝集素-2並非直接抑制T 細胞的增生,可能是藉由影響其他細胞激素來抑制T細胞的增生,先前研究指出半乳糖凝集素-1與介白素-10的合成有關,而我們利用介白素-10剔除小鼠的T細胞和一般小鼠的B細胞養出不會產生介白素-10的Treg-of-B細胞,經實驗發現剔除介白素-10的Treg-of-B細胞同樣具有抑制能力,而細胞相較於一般Treg-of-B細胞會表現較高的半乳糖凝集素-1,半乳糖凝集素-2和半乳糖凝集素-12,由此實驗結果可知,在沒有介白素-10的情況下,Treg-of-B細胞可能會轉錄更多的半乳糖凝集素基因,不過介白素-10和半乳糖凝集素之間的關係以及之間的機轉仍有待後續研究。 | zh_TW |
dc.description.abstract | Regulatory T cells induced by B cells (Treg-of-B cells) are a subpopulation of regulatory T cells. They are different from other regulatory T cells. Compared to natural regulatory T cells, Treg-of-B cells are Foxp3 negative, and they are CD25 positive while type 1 regulatory T cells (Tr1 cells) are CD25 negative. However, Treg-of-B cells suppressive function is not clearly understood, so we aim to further investigate how Treg-of-B cells suppress immune responses. Galectins are -galactoside proteins, and there are many researches showing that galectins play roles in innate and adaptive immunity. For example, galectin-2 suppresses immune responses by inducing CD8+ T cells apoptosis. However, in innate immunity, galectin-2 induces a proinflammatory phenotype in monocytes and macrophages.
We focused on the roles of galectins in Treg-of-B cells. We first checked the galectins gene expression in Treg-of-B cells by real-time PCR and found that galectin-2 was highly expressed in Treg-of-B cells while natural regulatory T cells express similar level as CD4+CD25- T cells. We further investigated the suppressive mechanism by blocking the possible binding partner of galectin-2, CD29. However, it showed no difference between the experimental group and control group. To check the blocking efficiency of anti-CD29, we found that galectin-2 could not suppress T cell directly, which suggested that galectin-2 might interact with other molecules to suppress T cell proliferation. It was reported that galectin-1 contributed to IL-10 synthesis. In our study, we found that not only galectin-1 mRNA expression increase, galectin-2 also express higher when IL-10 was deficient. More researches should be performed to clarify the correlation between galectins and IL-10. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T11:43:26Z (GMT). No. of bitstreams: 1 ntu-105-R03449008-1.pdf: 26627366 bytes, checksum: 20e74500dbb844fc47fcfc105a7feae7 (MD5) Previous issue date: 2016 | en |
dc.description.tableofcontents | 致謝……………………………………………………………………………i
中文摘要…………………………………………………………………..iii Abstract………………………………………………………………….iv I. Introduction 1 1. Background 2 1.1 B cell 2 1.2 T cell 3 1.3 Natural regulatory T cell (nTreg) 5 1.4 Induced regulatory T cell (iTreg) 7 1.5 Regulatory T cells induced by B cells (Treg-of-B) 8 1.6 Galectins 9 2. Motivation 10 II. Materials and methods 12 1. Materials 13 1.1 Mice 13 1.2 Medium 13 1.3 Buffer 13 1.4 EasySepTM system 14 1.5 BD IMagTM system 15 1.6 Mitogens, monoclonal antibodies and cytokines 15 1.7 Flow cytometry 16 1.8 Suppression test 17 1.9 RNA extraction 17 1.10 Reverse transcription polymerase chain reaction (RT-PCR) 18 1.11 Real-time polymerase chain reaction (quantitative PCR, qPCR) 18 1.12 Western blotting 20 2. Methods 22 2.1 Collection of primary cells 22 2.2 Treg-of-B cell culture 24 2.3 Type 1 Regulatory T cell culture 25 2.4 Flow cytometry 26 2.5 Suppression test 27 2.6 RNA extraction 27 2.7 Reverse transcription polymerase chain reaction(RT-PCR) 28 2.8 Real-time polymerase chain reaction (quantitative PCR, qPCR) 28 2.9 Western blot 29 III. Results 32 1.The purity of the splenocytes separated by magnetic beads 33 2.The surface molecules of Treg-of-B cells 33 3.The expression of forhead box P3 (Foxp3) of Treg-of-B cells 34 4.The suppressive function of Treg-of-B cells 34 5.The galectin profiles of natural regulatory T cells and Treg-of-B cells 35 6.The galectin expression of Treg-of-B cells and type 1 regulatory T cells 35 7.The galectin expression of CD4+CD25- T cells (responder T cells) 36 8.Galectin-2 secretion by Treg-of-B cells 36 9.CD29 expression on CD4+CD25- T cells 37 10.The suppressive function of Treg-of-B cells with CD29 blockade 38 11.The suppressive function of recombinant galectin-2 38 12.The suppressive function of IL-10 KO Treg-of-B cells 39 13.The galectin profiles of IL-10 KO Teg-of-B cells (compared to Treg-of-B cells)………… 39 IV. Discussion 40 V. Reference 45 Figures 50 | |
dc.language.iso | en | |
dc.title | 研究Galectins在Treg-of-B細胞的調控機轉上所扮演的角色 | zh_TW |
dc.title | The role of galectins in the regulatory function of Treg-of-B cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 104-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 賈景山(Jean-San Chia),楊宏志(Hung-chih Yang) | |
dc.subject.keyword | 調節性T細胞,半乳糖凝集素,CD29,介白素-10,T細胞抑制, | zh_TW |
dc.subject.keyword | Regulatory T cells,galectins,CD29,IL-10,T cell suppression, | en |
dc.relation.page | 64 | |
dc.identifier.doi | 10.6342/NTU201602654 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2016-08-15 | |
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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