Lee Ho-Bin, Piao Da-Chuan, Lee Jun-Yeong, Choi Jae-Yun, Bok Jin-Duck, Cho Chong-Su, Kang Sang-Kee, Choi Yun-Jaie
Department of Agricultural Biotechnology, Seoul National University, Seoul, 115-921, Republic of Korea.
Institute of Green-Bio Science and Technology, Seoul National University, 1447-1 Pyeongchang-Daero, Daehwa-Myeon, Pyeongchang-Gun, Gangwon-Do, 25354, Republic of Korea.
Microb Cell Fact. 2017 Feb 22;16(1):33. doi: 10.1186/s12934-017-0648-2.
Concerns regarding the safety of inactivated foot-and-mouth disease (FMD) vaccine have been raised since it is produced from cultured live FMD virus (FMDV). To overcome this issue, recombinant protein has been studied as an alternative vaccine.
We designed a chimerical multi-epitope recombinant protein (5BT), which is comprised of tandem repeats of five B cell epitopes (residue of VP1 136-162) derived from different FMDV variants and one T-cell epitope (residue of 3A 21-35). To increase solubility and stability of 5BT, it was conjugated with BmpB, the membrane protein B of Brachyspira hyodysenteriae (B5BT). Our results indicated that 5BT was susceptible to degradation by host protease and produced with substantial fraction of inclusion body. The stability and solubility of 5BT was greatly increased by conjugating to BmpB. FMDV specific antibodies were observed in the serum of mice immunized with 5BT and B5BT comparable to inactivated FMD vaccine. Sera from 5BT and B5BT groups also exhibited high epitope-specific antibody titers in peptide specific ELISA, indicating that all five epitopes are exposed to the B cell receptor for the antibody reaction. Thus the multi-epitope recombinant protein designed in this study may be a potential candidate as an alternative vaccine against FMDV epidemic variants.
由于灭活口蹄疫(FMD)疫苗是由培养的活口蹄疫病毒(FMDV)生产的,因此人们对其安全性表示担忧。为克服这一问题,重组蛋白已作为一种替代疫苗进行研究。
我们设计了一种嵌合多表位重组蛋白(5BT),它由来自不同FMDV变异株的五个B细胞表位(VP1 136 - 162残基)和一个T细胞表位(3A 21 - 35残基)的串联重复序列组成。为提高5BT的溶解性和稳定性,将其与猪痢疾短螺旋体的膜蛋白B(BmpB)进行偶联(B5BT)。我们的结果表明,5BT易被宿主蛋白酶降解,并产生大量包涵体。通过与BmpB偶联,5BT的稳定性和溶解性大大提高。在用5BT和B5BT免疫的小鼠血清中观察到FMDV特异性抗体,与灭活FMD疫苗相当。在肽特异性ELISA中,5BT和B5BT组的血清也表现出高表位特异性抗体滴度,表明所有五个表位都暴露于B细胞受体以进行抗体反应。因此,本研究设计的多表位重组蛋白可能是针对FMDV流行变异株的替代疫苗的潜在候选物。