2013-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/650090摘要：蚊蟲傳播登革熱過程除了吐出登革病毒也將唾液注入宿主。已知蚊唾液具備抗凝血因子、抗血小板凝集因子、血管舒張物質、醣類分解酵素、溶菌素和過敏免疫調控因子等，然多數功能尚屬未知。為瞭解蚊唾液與登革病毒感染間之交互關係，及其宿主之免疫反應，本研究將於第一年利用蚊蟲唾液體學資料庫和蚊基因體資料庫，分析病媒蚊受到登革病毒感染時，其特異性唾液蛋白和基因的表現差異。藉由基因沉寂、定量 RT-PCR 和蛋白質電泳等分析其唾液 RNA、唾液蛋白質與登革病毒增殖的關係。第二年，建立桿狀病毒表達系統生產特異性唾液重組蛋白，以細胞和病毒培養方法釐清唾液對登革病毒感染和複製過程中的影響。以小鼠模式比較蚊叮咬和皮下注射特異性重組蛋白後的唾液抗體動態變化。第三年，建立登革病媒具鑑別力的特異性唾液蛋白標記，以作為病媒叮咬和登革感染的指標。利用斑蚊、家蚊和瘧蚊等進行小鼠叮咬試驗，釐清蚊屬間唾液蛋白之交叉反應。第四年，分析登革病人血液中蚊唾液抗體強度與登革感染嚴重度之關係，以及登革流行區潛藏之蚊唾液抗體流行病學特性。登革病媒蚊之唾液蛋白組成、登革病毒感染和登革病人免疫反應之參考資料有限，本計畫將釐清上述蚊唾液在登革病毒感染過程中所扮演之角色。<br> Abstract: During blood feeding, mosquitoes infected with dengue virus inject the virus along with their saliva into the host. Mosquito saliva consists of anti-clotting factors, anti-platelet factors, vasodilators, glycolytic enzymes, lysozymes, and immunomodulatory factors; however, the functions of most salivary components remain unknown. To better understand the interactions among mosquito saliva, dengue virus infection, and the immune responses of the host, we will employ mosquito sialomic and genomic databases to study the expression of specific salivary proteins in response to dengue virus infection in the first year of the proposed study. The regulation of salivary proteins by dengue virus infection will be confirmed by gene silencing, quantitative RT-PCR, and protein electrophoresis analyses. In the second year, a baculovirus overexpression system will be used to produce specific recombinant salivary proteins, and the influence of mosquito saliva components on dengue virus infection will be investigated. The kinetics of antibody responses to mosquito saliva will be evaluated in a mouse model using mosquito bites as well as subcutaneous injection of the recombinant proteins. In the third year, indicators of vector exposure and dengue infection will be established for specific salivary proteins. The cross-reactivity of saliva from different Culicidae groups, e.g., Aedes spp., Culex spp., and Anopheles spp., will also be clarified using the mouse model. In the fourth year, the relationship between levels of antibodies against mosquito saliva and disease severity will be analyzed using serum samples collected from dengue patients. These studies will reveal the underlying epidemiological characteristics of anti-mosquito antibodies during dengue outbreaks. Limited information is currently available on the composition of the mosquito vector's saliva, dengue viral infection, and patient immune responses. This project will elucidate the role of mosquito salivary proteins in dengue infection.蚊蟲唾液蛋白登革熱mosquitosalivary proteindengue