Mannan conjugate
刺激 U937 細胞基因表現之探討
人類巨噬細胞 (macrophage) 的 mannose receptor ，在感染時期，對病原
體的偵測和反應是相當重要的。 mannose receptor 可以直接和病原體結
合而仲介吞噬作用 (phagocytosis) ，引發許多 cytokines 分泌調節免疫
系統，也可以增加活性氧中間產物的產生，增強細胞毒殺作用。過去
的研究，已經對於 mannose receptor 的基因、蛋白質結構有了深入的探
討，但對於 mannose receptor 所影響的分子層次方面，了解不多，甚至
對於 mannose receptor 所引起吞噬作用的訊號來源，也不是很清楚。近
年來，發展出許多 high-throughput 的研究方法，得以讓研究人員以系
統生物學的角度窺伺細胞運作的全貌，本篇論文中，依照 mannose rec
eptor 和 ligands 之間的結合特性，製備了 mannan conjugate 為刺激劑，
利用微矩陣列 (oligonucleotide microarray) 的技術分析出共有 5091 個基
因受到影響，並使用 real-time quantitative reverse-transcription polymera
se chain reaction (RT-PCR) 的方法，加以佐證結果的正確性，得到 80﹪
的一致性；此外，以二維電泳 (two-dimensional electrophoresis) 分析方
式，觀察細胞蛋白質表現變化，並進一步使用 ESI-Q-Tof 鑑別蛋白質
，希望能夠藉由 mannosylated glycoconjugate 和 mannose receptor 交互
作用下對基因、蛋白質表現的影響，而對 mannose receptor 在生物系統
上的角色有更深一層的認識。
Gene expression profile of U937-cell stimulated by mannan conjugate.
During infection, mannose receptor of human macrophages play a important role in pathogenic detection and responses. Macrophages have a lot of effector functions i nduced via mannose receptor, including mediated phagocytosis of pathogens, induc ed cytokines secretion to regulate immune system, and increased reactive oxygen in termediates production to enhance cytotoxicity. In past several years, there are man y researches about gene and protein structure of mannose receptor, but its molecula r effects, even the initiated signal(s) of phagocytosis events, are not identified yet.
Currently, advancement in methodology, researchers allows using high-throughput
methods to investigate behaviors of the cell by systematic biology. In the present st
udy, mannan conjugates was applied as mannose receptor ligands to allow us to exa
mine effected genes by oligonucleotide microarray. A total of 5091 genes were alte
red in the mannan conjugate treatment as compared with control. The results were
verified using real-time quantitative reverse-transcription polymerase chain reactio
n (RT-PCR). 80% consistence was acquired. Cellular protein expression level was f
urther analyzed by two-dimensional electrophoresis and identified by ESI-Q-Tof. B
ased on the interaction between mannose receptor and mannosylated glycocojugate
s, as an approach details about the effect of mannose receptor on expression of gene
s and proteins, as well as the biological mechanism, are hopefully more well unders
tood.
