出版年:

研究生:

楊世婷

研究生(英文姓名):

Shih-Ting Yang

論文名稱:

登革病毒感染小鼠細胞

英文論文名稱:

Infection of murine cells by dengue virus

指導教授:

黎煥耀

指導教授(英文姓名):

Huan-Yao Lei

學位類別:

碩士

校院名稱:

國立成功大學　

系所名稱:

微生物及免疫學研究所

學年度:

90

語文別:

中文

論文頁數:

65

關鍵詞:

第一型干擾素 ; 小鼠馴化登革二型病毒 ; 感染 ; 源自小鼠骨髓樹狀突細胞

英文關鍵詞:

Bone marrow-derived dendritic cells (BMDC) ;
Mouse adapted dengue virus type 2 (mDV2) ;
Type 1 Interferons (Type 1IFNs) ;
Infection

被引用次數:

0

[ 摘要 ]

登革熱是一種由蚊子媒介傳染的急性發作但可自然痊癒的病毒性傳染病，登革病毒感染有可能是類似感冒發燒造成較輕微的登革熱或具威脅性會致命的登革出血熱/登革休克症狀，目前並無有效抗病毒治療法及疫苗。最近登革病
毒目標細胞的研究顯示，單核球/巨噬細胞、神經細胞、肝臟細胞、內皮細胞、血球細胞株、週邊血液B淋巴細胞以及樹狀突細胞，都可能是登革病毒感染的目標，至於受感染的目標細胞在登革病毒致病機轉中所扮演角色及其影
響仍究不明。由於樹狀突細胞在病毒感染產生免疫反應的機制中扮演重要的調控角色，故其活體內是否可被登革病毒感染是相當重要的課題。我們以顆粒球/巨噬細胞聚落刺激因子培養源自骨髓小鼠樹狀突細胞，再以登革二型病
毒感染之，發現感染後24小時只在以OptiPrepTM梯度純化的低密度樹狀突細胞族群中測得代表病毒正在複製的負股核酸及病毒抗原。顯示只有極少數細胞被感染且容許登革二型病毒複製，我們同時測得樹狀突細胞產
生大量α干擾素可能導致病毒不易在其內複製。為模擬蚊子叮咬皮膚自然感染的情形，我們將登革二型病毒直接皮下給予腳掌，觀察皮膚組織特化樹狀突細胞可否被登革病毒感染或擷取病毒抗原而移行至淋巴結藉此感染其他細胞
。發現淋巴結懸浮液以OptiPrepTM梯度純化分離，低密度細胞族群中CD11c+Gr-1-細胞約佔40~50%為樹狀突細胞，13、17小時各測得約0.01%的細胞表現登革核心蛋白，除為CD205+樹
狀突細胞外，也觀察數顆病毒蛋白陽性反應並不表現CD205的細胞。為增加小鼠被登革病毒感染我們於腳掌給予能生產病毒的小鼠肝癌細胞株，12、18小時於腿部淋巴結切片也偵測到登革病毒核心蛋白、外套膜蛋白表現

[ 英文摘要 ]

Dengue virus (DV), an arthropod-borne flavivirus, causes a febrile illness or severe hemorrhagic fev
er for which there is no effective antiviral treatment and no vaccine available. It is still not cle
arly understood which tissue cells are the natural targets of DV. In addition to monocytes/ macropha
ges, neuron, hepatocytes, endothelial cells, B lymphcytes and dendritic cells have also been reporte
d as potential targets for DV. Recent studies have demonstrated two subsets of dendritic cells, mono
cyte-derived dendritic cells and human skin Langerhans cells are targets for dengue virus infection.
We are interested whether mouse dendritic cells can be infected by DV. Large numbers of dendritic c
ells from mouse bone marrow cultures supplemented with Granulocyte/ Macrophage Colony-stimulating Fa
ctor (GM-CSF) were generated. We found that bone marrow-derived dendritic cells (BMDC) can be infect
ed with dengue virus. At 24 hours post-infection, negative- and positive-strand viral genome was det
ected on low density BMDC by RT-PCR, viral antigens were also detected by immunofluorescence analysi
s. The efficiency of DV infection on BMDC was extremely low, because BMDC express Type 1 Interferon.
Furthermore, DV was injected subcutaneously into mouse skin to mimic mosquito bite. The spread of D
V from inoculation site to the draining lymph node was characterized. The lymph node dendritic cells
were isolated by optiprep gradient and the viral antigen was analyzed by immunofluorescence assays.
The efficiency of DV infection on mouse cells was extremely low. A mouse hepatoma cell ML-15a can s
upport dengue virus replication. This DV2 infected-ML-15a cell was inoculated into mouse footpad. DV