開發以鞭毛蛋白為佐劑之豬流行性下痢病毒不活化疫苗

豬流行性腹瀉病毒 (PEDV) 是對所有年齡豬都具有高度傳染性的病源，此病毒對養豬業和肉類市場造成了嚴重的經濟損失，而預防病毒性疾病的最佳方法是接種疫苗。本研究以滅活的PEDV為抗原，結合鞭毛蛋白佐劑，進行口服疫苗製備與測試的研究。口服疫苗具有許多優點，主要為易於儲存和接種、並能誘發黏膜性免疫反應。為了解決滅活PEDV抗原在口服疫苗使用上的局限性，並提高免疫反應，本研究嘗試利用添加佐劑來改善。鞭毛蛋白是沙門氏菌尾部的一種成分，它可以激活 Toll 樣受體5 (TLR5)以激活促炎免疫反應。在這項研究中，鞭毛蛋白被用作佐劑以增強免疫效力。鞭毛蛋白在大腸桿菌BL21表達系統中表達後進行純化，並經過SDS-PAGE及Western Blot分析驗證成功。接著，為了保護抗原和鞭毛蛋白佐劑免受小鼠胃部消化，抗原及佐劑利用褐藻酸鈉進行包覆。最後利用小鼠實驗測試口服疫苗的效果，在IgG和IgA評估中顯示鞭毛蛋白的添加確實能夠增強IgA的效果，顯示此口服疫苗在誘發黏膜保護的成果。

關鍵字

豬流行性腹瀉病毒；鞭毛蛋白；佐劑；滅活病毒；口服疫苗

並列摘要

Porcine Epidemic Diarrhea Virus (PEDV) is a cause of highly contagious diseases in all ages of pigs. It was causing severe economic damage to the pig breed industry and the meat market. The best way for prevention measure is vaccination. In this study, the inactivated PED virus was used as an antigen combined with the Flagellin protein adjuvant for oral immunization. Besides many advantages, including low budget price, short research and development time requirement, and easy storage and vaccination. The limitations of the inactivated vaccine against the PED virus problem needed to be solved to improve immune response; it could be improved by adjuvant. Flagellin is a component from the tail of Salmonella; it can activate Toll-like receptor 5 (TLR5) to activate pro-inflammatory immune responses. In this study, Fagellin was used as an adjuvant to enhance immune effectiveness. Flagellin was expressed in E.coli BL21 expression system and analyzed by SDS-PAGE, and Western Blot used His-Taq Antibody. Furthermore, to protect the antigen and flagellin adjuvant against the digestion on mice’s intestine, they were coated with Alginate before vaccination. The immune evaluation to confirm the effectiveness of adjuvant are direct ELISA for IgG and IgA evaluation.