汉滩病毒结构蛋白昆虫细胞融合表达产物的免疫原性评价

Evaluation of immunological properties of fused expression product of Hantaan virus structural proteins in insect cells

目的 研究汉滩病毒 (HTNV)囊膜糖蛋白G1与核蛋白 (NP)部分片段在昆虫细胞中融合表达产物的免疫学特性 ,为HTNV基因工程疫苗的研制提供依据。方法 构建含有HTNVG1S0 .7嵌合基因的重组杆状病毒Bac G1S0 .7,用其感染的Sf9细胞免疫Balb c小鼠。用间接免疫荧光法、ELISA法、微量细胞培养中和试验及T淋巴细胞增殖试验对被免疫小鼠的体液免疫及细胞免疫应答效果进行检测。结果 Bac G1S0 .7感染的昆虫细胞免疫小鼠后 ,测得免疫鼠血清抗HTNV抗体滴度最高达 1∶3 2 0 0 ,含有特异性抗HTNVNP及抗HTNV糖蛋白G1的抗体 ,被免疫小鼠中有部分产生了较低滴度的中和抗体 ,免疫鼠脾细胞对HTNVNP或糖蛋白的增殖反应指数均高于阴性对照组。结论 G1S0 .7嵌合基因的昆虫细胞表达产物具有较好的免疫原性 。

Objective To study the immunological properties of fused expression product of Hantaan virus glycoprotein G1 and nucleoprotein partial fragment in insect cells, and lay the foundation for developing Hantaan virus genetic engineering vaccine.Methods The recombinant baculovirus Bac G1S0.7 with chimeric gene G1S0.7 of Hantaan virus was constructed. Mice were injected with Sf9 cells infected by the recombinant baculovirus. The humoral and cellular immunological effects were identified by indirect immunofluorescence, ELISA, micro cell culture neutralization test and T lymphocytes stimulation test.Results Immunized by Bac G1S0.7 infecting insect cells, specific antibody with the highest titre of 1∶3 200 was observed, which could specifically bind to NP and glycoprotein of Hantaan virus. Low neutral antibody titres were tested in part of the immunized mice. The stimulation indexes of splenocytes of immunized mice to NP and glycoprotein of Hantaan virus were all higher than those found in the negative control.Conclusions The expression product of chimeric gene G1S0.7 in insect cells is immunogenic, which can simultaneously stimulate humoral and cellular immunological responses against glycoprotein G1 and NP of Hantaan virus.