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  • 學位論文

腺病毒於禽流感M2疫苗發展之評估

Evaluation of M2-based avian influenza virus vaccine using an adenovirus vector

指導教授 : 王金和
共同指導教授 : 鄭益謙(Ivan-Chen Cheng)

摘要


一般商業化之家禽流行性感冒病毒之疫苗對於不同HA亞型之家禽流行性感冒病毒感染無交叉保護力。因此,本實驗以發展具交叉保護之疫苗為目的,利用複製缺陷腺病毒 (replication deficient adenovirus) 作為載體,建構帶有不同亞型間保守度高之M2基因片段的重組腺病毒 (Ad/M2),再將Ad/M2免疫雞隻。Ad/M2感染之293A細胞及轉導之Vero細胞以間接螢光免疫分析皆有陽性訊號產生。在Ad/M2免疫雞隻的實驗結果顯示,補強後2週CD4+及CD8+ T cell相較於初次免疫有顯著增加;抗M2特異性抗體則於初次免疫後2週有顯著增加,但補強後的抗體上升效果則不明顯。於攻毒後第4天及第7天採集的喉頭拭子之real time RT-PCR結果顯示免疫組與對照組之間的AIV含量沒有差異,而攻毒後免疫組與對照組死亡率分別為40%及33.3%,給予Ad/M2之組別並無法對抗AIV感染。

並列摘要


Current vaccines to avian influenza virus can’t induce broad cross-protection against different subtypes. Therefore, the purpose of this research was to develop a vaccine which can confer cross-protection by using replication-deficient adenovirus as a vector to construct a recombinant adenovirus carrying highly conserved M2 gene (Ad/M2). The immunofluorescence assay revealed positive signal in 293A and Vero cells transduced with Ad/M2. The Ad/M2 was intranasally and intramuscularly immunized in chickens twice at a 4-week interval. Anti-M2 specific antibody was significantly higher after two weeks post primary vaccination. Challenging with AIV after two weeks post boost vaccination, the consequence of real time RT-PCR revealed AIV content was no difference between Ad/M2 group and control group in oropharyngeal swab on day 4 and day 7 post challenge. The mortality of Ad/M2 group and control group was 40% and 33.3% respectively. The conclusion is giving chickens Ad/M2 couldn’t confer protection against AIV infection.

參考文獻


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