Li Long, Fang Weihuan, Li Jianrong, Huang Yaowei, Yu Lian
Institute of Protective Veterinary, College of Animal Science, Zhejiang University, Hangzhou, 310029, PR China.
Vaccine. 2006 Aug 14;24(33-34):5919-27. doi: 10.1016/j.vaccine.2006.04.057.
Our previous study showed that vaccination with plasmid DNA containing infectious bursal disease virus (IBDV) gene which encodes complete polyprotein (VP2/4/3) induced protective immune responses. In this study, we examined the efficacy of an oral DNA vaccine carrying the IBDV polyprotein antigen delivered by attenuated Salmonella enterica sv. Typhimurium (S. typhimurium). The recombinant plasmid pCI-VP2/4/3 was transformed by electroporation into an attenuated S.typhimurium Strain (Dam Phop) (designated hereafter as SV/pCI-VP2/4/3). The IBDV polyprotein gene was expressed in chicken embryo fibroblast (CEF) cells infected with strain SV/pCI-VP2/4/3, as shown by gene-specific RT-PCR and Western blot. Oral immunization of 7-day-old specific-pathogen-free (SPF) chickens with SV/pCI-VP2/4/3 elicited specific humoral responses as measured by ELISA. Vaccination with the strain SV/pCI-VP2/4/3 at 10(9) CFU per chicken offered 11/15 (73%) protection of the chickens against virulent IBDV challenge. Our results have implications in the development of DNA vaccines against avian viral diseases by bacteria-vectored oral delivery system.
我们之前的研究表明,用含有传染性法氏囊病病毒(IBDV)基因(编码完整多聚蛋白(VP2/4/3))的质粒DNA进行疫苗接种可诱导保护性免疫反应。在本研究中,我们检测了由减毒肠炎沙门氏菌鼠伤寒亚种(鼠伤寒沙门氏菌)递送的携带IBDV多聚蛋白抗原的口服DNA疫苗的效力。通过电穿孔将重组质粒pCI-VP2/4/3转化到减毒鼠伤寒沙门氏菌菌株(Dam Phop)中(以下称为SV/pCI-VP2/4/3)。如基因特异性逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法所示,IBDV多聚蛋白基因在感染菌株SV/pCI-VP2/4/3的鸡胚成纤维细胞(CEF)中表达。用SV/pCI-VP2/4/3对7日龄无特定病原体(SPF)鸡进行口服免疫,通过酶联免疫吸附测定(ELISA)检测到特异性体液反应。每只鸡用10⁹CFU的菌株SV/pCI-VP2/4/3进行疫苗接种,为11/15(73%)的鸡提供了针对强毒IBDV攻击的保护。我们的结果对通过细菌载体口服递送系统开发抗禽病毒病DNA疫苗具有启示意义。