重组杆状病毒表达的EIAV Env蛋白与含有env基因重组痘苗病毒的联合免疫

目的：构建新型的马传染性贫血病毒(EIAV)的候选疫苗：方法：利用BAC—To—BAC杆状病毒表达系统，将中国马传贫驴白细胞弱毒疫苗(EIAV DLV)及其亲本株(EIAV LN)env基因导入到杆状病毒基因组中。转染昆虫细胞后，得到的重组病毒用SDS—PAGE和Western blot检测表达产物。以本实验室构建的含有EIAV env基因的重组痘苗病毒，单独或与重组杆状病毒表达的EIAV Env蛋白联合免疫小鼠。结果：构建的重组杆状病毒能正确表达全长Env蛋白。与单独免疫组相比，联合免疫组免疫应答显著增强，其中中和抗体的滴度提高5～9倍。结论：含有EIAV env基因的重组痘苗病毒与Env蛋白抗原联合免疫，能够诱导高滴度的中和抗体。

Combination immunization with EIAV Env protein expressed by recombinant baculovirus and recombinant vaccinia virus containing env gene

AIM: To develop a novel vaccine candidate of Equine infectious anemia virus(EIAV). METHODS: env genes of EIAV Chinese donkey leukocyte attenuated strain (EIAV DLV) and its parental virus strain (EIAV LN) were expressed using the BAC-To-BAC system, and Env proteins were confirmed by SDS-PAGE and Western blot. BALB/c mice were immunized with recombinant vaccinia viruses containing env genes of EIAV alone or boosted with Env proteins expressed by recombinant baculovirus. Both protective humoral and cellular immune responses were detected. RESULTS: Recombinant baculovirus could express complete Env proteins and the titer of neutralizing antibody in the prime-boost group was found to be 5 to 9 folds higher than that in the rVV group. CONCLUSION: Combination of protein antigen and recombinant vaccinia virus could induce high titer of neutralizing antibody against EIAV. This work facilitates development of a novel genetically engineered vaccine of EIAV as a model for future development of an HIV vaccine.