呼吸道融合瘤病毒(Respiratory Syncytial virus, RSV)為副黏液病毒科(Paramyxoviridae)成員之一，會感染幼兒、老年人及免疫低下的個體，造成嚴重的下呼吸道感染。美國每一年超過九萬的幼兒感染而住院治療。但是截至目前為止仍無臨床上認證的疫苗。
腺病毒載體是近年來熱門且吸引人的疫苗研究方向，可提升體液免疫反應及細胞免疫反應。本研究擬以呼吸道融合瘤病毒外套膜醣蛋白F基因為標的基因，研究腺病毒載體做為疫苗的效力。首先將呼吸道融合瘤病毒外套膜醣蛋白F 基因全長以及擁有不同功能區塊的跨模截斷蛋白基因構築至腺病毒載體(Ad-F0，Ad-F0ΔTM)；之後感染293A細胞48小時後，抽取細胞RNA以RT-PCR進行RNA表現測定，顯示腺病毒載體皆有進行基因轉錄作用。
動物實驗方面，將BALB/c以及C57BL/6小鼠鼻腔內注射各重組腺病毒載體疫苗，隔10天或20天皮下追加注射一劑，並於第10、20、30天進行採血，測定抗體效價反應及中和試驗以及分離脾臟細胞測定細胞激素IFN-γ、IL-2和IL-4分泌的情形，結果顯示免疫接種載體Ad-F0ΔTM，Ad-F0的組別，血清對於病毒之抗體效價(ELISA) 較為顯著；其病毒中和性抗體產生亦有明顯反應，追加免疫後，血清效價上升更為顯著；並且測定血清之同型抗體，結果顯示Ad-F0的組別可能趨向Th1免疫反應。在細胞免疫方面，免疫接種Ad-F0的組別，用CD8+、CD4+以及RSV F蛋白25個區段抗原性胜肽刺激，脾臟細胞增加分泌細胞激素IL-2和IFN-γ。
根據上述實驗結果，我們已經成功構築腺病毒載體疫苗，能夠引起對抗呼吸道融合瘤病毒的免疫反應。日後研究會著重於引起黏膜性反應，以達到發展老年人及小孩的呼吸道融瘤合病毒疫苗。

Respiratory Syncytial Virus (RSV), a member of the Paramyxoviridae, can cause severe lower respiratory tract infections in children, the elderly and immune-compromised individuals. More than 90,000 children in the US are hospitalized every year as a result of this viral infection. But there is no clinically licensed, effective RSV vaccine.
Adenovirus vector is a vaccine developing field. It can elicit persistent humoral and cellular immune responses. The aim of this study was to develop the recombinant vaccine: Adenoviral vectors carrying RSV F genes as vaccines. Therefore we construct RSV- full-length F0 and transmembrane truncated F0ΔTM glycoprotein transgene into the adenovirus type 5 vectors (Ad-F0, Ad-F0ΔTM ). We detected the F0 and F0ΔTM expression in 293A cell culture model. After 48 hours’ transfection, we extracted total cellular RNA and detected equivalent gene expressions. The result showed that all of two recombinant vectors could express efficiently.
In animal study, the female BALB/c or C57BL/6 mice were immunized with Ad-F0, Ad-F0ΔTM, Ad-LacZ, live RSV or inactivated RSV by intranasal injection. Booster was performed after 10 or 20 days by subcutaneous injection. Blood samples for serology assay were obtained by tail bleeding on day 10, 20 and 30. The results showed that mice immunized with Ad-F0ΔTM and Ad-F0 vectors had better antibody responses. After final booster inoculation, serum antibody titers were further upgraded and virus-neutralizing antibody titers were also dominantly detected. Ad-F0-vaccinated mice presented the switching of the ratio of IgG2a/ IgG1 specific antibody responses in the sera indicating the activation of Th1 immunity.
In cellular immune response, the results showed that mice immunized with Ad-F0 vectors had increased IL-2 and IFN-γ secretion upon the stimulation of CD8+, CD4+ peptides or 25 RSV-F regions peptides mixtures.
According to the presented results, we had successfully constructed adenoviral vector vaccines which can induce immune response against RSV. The further work, we will focus on mucosal immunity in order to develop the RSV vaccine for the elderly and children.

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