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Title: | 利用DNA微陣列晶片分析研究調節性T細胞的基因表現 Comparative Studies on the Gene Expression of Regulatory T Cells by DNA Microarray Analysis |
Authors: | Wen-Jan Chiang 姜文然 |
Advisor: | 江伯倫(Bor-Luen Chiang) |
Keyword: | 調節性T細胞,CD4+CD25+調節性T細胞,誘導型調節性T細胞,第一型調節性T細胞, Regulatory T cell (Treg),Naturally occurring CD4+CD25+Foxp3+ T cell (nTreg),Inducible regulatory T cells (iTreg),Type 1 regulatory T cell (Tr1), |
Publication Year : | 2008 |
Degree: | 碩士 |
Abstract: | 目前廣泛地認知在人類和齧齒類動物中有一群被命名為”調節性T細胞”的細胞族群具有調節免疫反應的能力,來維持自體耐受性和免疫系統的體內平衡。在過去幾年內,免疫學者研究指出在CD4+CD25+調節性T細胞裡,除了CD25分子標記外,還發現轉錄因子Foxp3在此種T細胞裡扮演著關鍵的角色,無論如何現今仍存在著許多關於這些調節性T細胞發育、分化和作用機轉,甚至是調節性T細胞彼此之間相互影響的問題等待解決。
因此在本實驗當中,我們使用帶有IL-10基因之重組慢病毒系統來感染T細胞,使得T細胞會大量表現IL-10基因並且轉變成為一種”類第一型調節性T細胞”,接著利用DNA微陣列晶片技術來進行基因表現分析,並且加以比較我們實驗室另外兩種以同樣系統表現Foxp3和TGF-β基因之微陣列晶片的結果進行綜合分析,進一步地挑選出標的基因進行實驗,觀察它們在活化或未活化的調節性T細胞裡的基因表現,另外我們從BALB/c小鼠中取出CD4+ T細胞和細胞激素TGF-β一起繼代培養,在加入TGF-β後的CD4+ T細胞可引起轉錄因子Foxp3之表現,轉變成為一種週邊誘導型調節性T細胞,並且同樣地去觀察標的基因在此種調節性T細胞裡的表現。 在我們初步分析微陣列晶片的結果中發現共有37個基因各自在兩種晶片的結果裡顯示有表現,經過篩選後挑出11個受到我們注意並想要進一步研究的基因,有Gsk3b, Ogt, Dub1, Ptprj, Ccr2, Lck, Icos, Ptprk, Slc2a1, Adm 和 Crem,另外值得注意的是我們發現其中在Foxp3和TGF-β基因之晶片結果當中各自有個具高度表現的基因,目前在一些研究中被認為可能是調節性T細胞的專一性標記,分別是Gzmb 和 Nkg7基因,然而在這些初步的分析結果後,還有待我們進一步地研究這些基因和調節性T細胞之間的關係。 It is widely known that a population of T cells named “regulatory T cells” (Tregs) might control the immune response in humans and rodents, maintain the self-tolerance and immune homeostasis. For the past few years, except for the surface marker CD25, immunologists found Foxp3 to be the key master factor in naturally occurring CD4+CD25+ T cells. However, there still have many questions about development, differentiation and mechanisms in the subset of Tregs even including their interaction between each other that need to be answered. Therefore, in our study, we exploited lentivirus system carrying IL-10 gene to infect T cells. Making T cells over-express IL-10 gene and converted it into an IL-10-secreting Tr1-like T cell. Following that, we used the DNA microarray technique for gene analysis and compared with the data of Foxp3- and TGF-β-transduced T cells in our laboratory, then selected some target genes for examination of gene expression both in fresh and activated Treg compared with non-Treg. In addition, we isolated naïve T cells (CD4+CD25-) from BALB/c mice and treated in the presence of TGF-β in vitro and the cells can be induced to express Foxp3, converted it into peripheral inducible Treg. We also investigated gene expression of the target genes in these TGF-β-derived Foxp3+ T cells. In our basic finding, there were 37 genes expression in both of the microarray data, and we narrowed down to 11 genes which we interest and further investigate, such as Gsk3b, Ogt, Dub1, Ptprj, Ccr2, Lck, Icos, Ptprk, Slc2a1, Adm and Crem. Notably, we also found that the two genes were highly expressed in the data of Foxp3- and TGF-β-transduced T cells, and have been considered as specific markers of natural Treg in several reports, such as Gzmb and Nkg7. However, there is still needed to further investigate the relationship between these genes and nTreg. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40117 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 口腔生物科學研究所 |
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