- 學位論文
利用腺病毒載體及第三區域套膜蛋白融合重組鞭毛蛋白進行prime-boost免疫法研發四價登革熱疫苗
Prime-boost immunization using adenovirus vector and envelope domain III protein fused with flagellin for tetravalent dengue vaccine development
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摘要
每年感染登革熱的病人有顯著增加的現象，顯示登革熱已經成為全球密切注意的健康問題。套膜蛋白中的第三區域是病毒與細胞受器結合的位置，此外第三區域蛋白也被證實是一段具有血清型特異性的片段。在此篇研究中，我們利用大腸桿菌製造出一個融合蛋白，此蛋白除了帶有膜蛋白的第三區域之外還具有TLR5的配體(鞭毛重組蛋白flagellin)，實驗證實在小鼠體內，這一種特殊的蛋白可以誘發顯著的中和性抗體。除此之外，我們結合這一種融合蛋白與帶有前膜蛋白和套膜蛋白序列的腺病毒載體來引誘更高的免疫反應，這一種方式稱為heterologous prime-boost strategy。我們發現在小鼠的血清中，第三區域特異性的抗體是主要提供中和能力的亞群，同時我們也偵測到heterologous prime-boost strategy可誘發顯著的第三區域特異性total IgG力價。最重要的是，heterologous prime-boost strategy可以引誘出針對四種血清型的中和性抗體。這些結果顯示以腺病毒載體與套膜第三區域片段組合而成的heterologous prime-boost strategy具有發展登革熱疫苗的可行性。
並列摘要
A dramatic increase in incidents has been documented indicating that DENV has become a major international health concern. E protein domain III (EDIII) is the receptor binding domain, and was demonstrated that EDIII contains serotype-specific epitopes. In this study, the E.coli-based fusion protein, containing a TLR5 agonist, flagellin (FliC), and EDIII epitopes in a single polypeptide, which could elicit significantly higher neutralizing antibodies than EDIII mixed with FliC. We further evaluated the heterologous prime-boost strategy using adenovirus vector encoding prME and FliC-EDIII fusion protein in BALB/c mice. We found anti-EDIII specific antibodies were the dominant neutralizing ability, while heterologous prime-boost strategy elicited significantly higher anti-EDIII total IgG titer compared with homologous prime-boost immunization. Moreover, the neutralizing ability against all serotypes of DENV was observed in tetravalent Ad-prME prime/FliC-EDIII boost regimen. These result demonstrated the feasibility of developing DENV vaccine using heterologous prime-boost strategy.
參考文獻
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