本文選題：6A8α-甘露糖苷酶 + Jurkat細胞�。� 參考：《中國協(xié)和醫(yī)科大學》2006年博士論文

【摘要】： 蛋白質糖基化在細胞的功能行為中起重要的作用。本實驗室以往克隆到了1個新的人α-甘露糖苷酶cDNA(6A8)，并觀察到了6A8α-甘露糖苷酶表達抑制影響包括Jurkat細胞在內的多種細胞的生物學行為。在預實驗中觀察到6A8α-甘露糖苷酶表達抑制的Jurkat細胞(AS)在培養(yǎng)中聚集成大的團塊，這提示AS細胞間的粘附性增強。為研究其機制，首先用含有粘附分子和其它免疫分子基因在內的440個基因的DNA芯片對AS和M細胞(轉導空載體的對照細胞)間的基因表達差異作了分析，見AS細胞有19個基因的表達上調，有20個基因的表達下調。表達上調的基因中包括CD11a(整合素α-L)、CD54(ICAM-1)、CD82、CD24、CD11c(整合素α-X)、整合素α-7、CD103(整合素α-E)、TNFSF9、IL-1R和IL-2Rγ等與細胞間粘附相關的基因。RT-PCR和免疫熒光染色支持芯片的結果。這些基因中以CD54和CD11a最為重要。本論文工作中對它們作了重點研究。與對照細胞(野生型細胞W與M細胞)相比，AS細胞與包被有CD54分子的培養(yǎng)板的粘附增強。封閉性CD11a單抗能阻斷細胞與包被有CD54分子的培養(yǎng)板的粘附及細胞間的粘附。這證明了CD54和CD11a表達增強參與AS細胞間粘附增強。AS細胞LFA-1分子的親和力也增高。另外，AS細胞間的粘附增強也與細胞骨架排列改變相關。異種細胞間的粘附在免疫應答等的生命活動中也起重要的作用。在對AS細胞與對照細胞和Raji細胞間粘附的比較中，觀察到前者和Raji細胞間的粘附增強。在超抗原SEB存在下，Jurkat細胞與Raji細胞間可形成免疫突觸，并有信號傳導發(fā)生。6A8α-甘露糖苷酶的功能是修剪N-糖鏈中的甘露糖，Con A結合實驗能檢測細胞糖蛋白N-糖鏈中的甘露糖被α-甘露糖苷酶修剪的程度。與對照細胞相比，Con A與AS細胞的結合明顯增強。關于6A8α-甘露糖苷酶表達抑制所致的蛋白質糖基化改變影響Jurkat細胞粘附性的確切機制須作深入研究。
[Abstract]:Glycosylation of proteins plays an important role in the functional behavior of cells. A new human 偽 -mannosidase cDNA (6A8) was cloned in our laboratory, and the inhibition of the expression of 6A8 偽 -mannosidase was observed to affect the biological behavior of many cells, including Jurkat cells. In the pre-experiment, Jurkat cells (as), whose expression of 6A8 偽 -mannosidase was inhibited, were observed to aggregate into large lumps in culture, which indicated that the adhesion between as cells was enhanced. In order to study its mechanism, the difference of gene expression between as and M cells (control cells of empty vector) was analyzed by DNA microarray containing 440 genes including adhesion molecules and other immunomolecular genes. There were 19 genes up-regulated and 20 genes down-regulated in as cells. The up-regulated genes included CD11a (integrin 偽 -L), CD54 (ICAM-1), CD82, CD24, CD11c (integrin 偽 -X), integrin 偽 -7CD103 (integrin 偽 -E), TNFSF9IL-1R, IL-2R 緯 and other genes associated with intercellular adhesion. Of these genes, CD54 and CD11a are the most important. In this paper, we focus on them. Compared with the control cells (wild-type cells W and M cells), the adhesion of as cells to the culture plates coated with CD54 molecules was enhanced. The closed CD11a McAb could block the adhesion of cells to the culture plate coated with CD54 molecule and the adhesion between cells. It is suggested that the increased expression of CD54 and CD11a may play an important role in the affinity of LFA-1 molecules in as cells. In addition, the enhancement of adhesion between as cells was also related to the change of cytoskeleton arrangement. Adhesion between xenogeneic cells also plays an important role in immune response and other life activities. The adhesion between as cells and Raji cells was observed in comparison with control cells and Raji cells. Immune synapses can be formed between Jurkat cells and Raji cells in the presence of superantigen SEB. The function of signal transduction. 6A8 偽 -mannosidase is to prune mannose Con A in N- sugar chain to detect the degree of mannose pruning by 偽-mannosidase in cell glycoprotein N- sugar chain. The binding of Con A to as cells was significantly increased compared with the control cells. The exact mechanism of the effect of 6A8 偽 -mannosidase expression inhibition on the adhesion of Jurkat cells needs further study.
【學位授予單位】：中國協(xié)和醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2006
【分類號】：R392

【參考文獻】

相關期刊論文 前2條

1 岳瑋,史耕先,王壯志,劉音,朱立平;轉導反向6A8α-甘露糖苷酶cDNA對抗Fas抗體誘導Jurkat細胞凋亡的影響[J];基礎醫(yī)學與臨床;2001年02期

2 李琳,王壯志,馬鳳蓉,史耕先,趙方萄,朱立平;6A8 α-甘露糖苷酶也表達于細胞膜[J];中華微生物學和免疫學雜志;2001年04期

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