開發牛病毒性下痢重組E2蛋白次單位疫苗

Development of recombinant E2 antigen subunit vaccine against bovine viral diarrhea virus

林玟因(Lin Wen-Yin)

指導教授 ： 鍾曜吉

國立屏東科技大學/獸醫學院/動物疫苗科技研究所/碩士(2018年)

https://doi.org/10.6346/THE.NPUST.AVT.008.2018.D03

摘要

牛病毒性下痢病毒（Bovine viral diarrhea virus, BVDV）是造成全世界牛群疾病的主要病原體，並對其對生產和一般健康狀況的不利影響造成重大經濟損失。 接種疫苗廣泛用於控制全世界BVDV感染的表現，但是BVDV野毒株的異質性和BVDV建立持續感染的獨特能力限制了疫苗的有效性。
在這項研究中，我們為台灣BVDV野生型構建了一種新的重組pET24a-BVDV-E2，其表達BVDV具有高免疫原性E2蛋白。使用大腸桿菌表達系統(Escherichia coli expression system)來表達這種重組蛋白並 Western blotting驗證E2蛋白的抗原性，然後將E2蛋白與ISA 206佐劑混合製成疫苗。我們已經確認注射小鼠疫苗的功效，雖然E2亞單位疫苗沒有達到預期的效果，但E2蛋白與少量滅活疫苗混合可提高滅活疫苗的有效性。 因此，我們推測rE2蛋白可能可以增強滅活BVDV疫苗效力。

關鍵字

牛病毒性下痢病毒 ； E2蛋白 ； 次單位疫苗

並列摘要

Bovine viral diarrhea virus (BVDV) is a major pathogen responsible for diseases in cattle population worldwide and causes substantial economic losses for its negative effect on production and general health conditions. Vaccination is widely used to control manifestations of BVDV infections in the world, but in the heterogeneity of BVDV field strains and the unique ability of BVDV to establish persistent infections, limiting the effectiveness of the vaccine. In this study, we have constructed a novel recombinant pET24a-BVDV- E2 for Taiwan BVDV wild-type expressing the most immunogenic E2 antigen of BVDV. This recombinant protein was expressed by Escherichia coli expression system. The expression of E2 protein was verified by Western blotting. And then the E2 protein was mixed with ISA 206 adjuvent to produce subunit vaccine. We have confirmed the efficacy of the vaccine by Injected mice, the E2 subunit vaccine does not deliver on expectations, But the E2 protein mixed with a small amount of inactivated vaccine improve the effectiveness of inactivated vaccines. Therefore, we speculate that rE2 protein perhaps be an effective way to enhance the protective efficacy of inactivated BVDV vaccines.

並列關鍵字

Bovine viral diarrhea virus ； E2 protein ； subunit vaccine

參考文獻

中華民國《動物用藥品檢驗標準》106年版.

Yesilbag K, Alpay G, Becher P. Variability and Global Distribution of Subgenotypes of Bovine Viral Diarrhea Virus. Viruses. 2017;9.

Cibulski S, Rivera-Patron M, Suárez N, Pirez M, Rossi S, Yendo AC, et al. Leaf saponins of Quillaja brasiliensis enhance long-term specific immune responses and promote dose-sparing effect in BVDV experimental vaccines. Vaccine. 2018;36:55-65.

Ridpath JF. Immunology of BVDV vaccines. Biologicals. 2013;41:14-9.

Tautz N, Meyers G, Thiel H-J. Pathogenesis of mucosal disease, a deadly disease of cattle caused by a pestivirus. Clinical and Diagnostic Virology. 1998;10:121-7.