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目的 研究基因重组表达的汉滩病毒核蛋白及其氨基端 2 6 ku片段在刺激机体免疫应答中的作用 ,为汉滩病毒基因工程疫苗研究打下基础 .方法 采用基因重组技术和亲和层析技术对汉滩病毒核蛋白及其氨基端 2 6 ku片段进行表达和纯化 ,并通过体外和体内试验对上述表达产物刺激小鼠免疫应答的作用及其特性进行了初步鉴定 .结果 汉滩病毒核蛋白及其 2 6 ku片段对小鼠均具有较强的免疫原性 .EL ISA检测免疫鼠抗体滴度可达 1∶ 32 0 0 0以上 ,并且均可刺激产生不同滴度 (1∶ 10～ 1∶ 16 0 )的中和抗体 .被动转移免疫鼠脾细胞对汉滩病毒感染乳鼠具有保护作用 ,其中 2 6 ku片段免疫鼠脾细胞的保护作用尤为明显 .结论 汉滩病毒核蛋白及其氨基端 2 6 ku片段能刺激机体产生中和抗体 ,同时可刺激机体细胞免疫应答 .
Objective To study the role of Hantaan virus nucleoprotein and its amino-terminal 26 ku fragment in stimulating the immune response of human beings and lay a foundation for the study of Hantaan virus genetically engineered vaccine.Methods Recombinant DNA and affinity chromatography Hantaan virus nucleoprotein and its amino-terminal 26 ku fragment for expression and purification, and in vitro and in vivo experiments on the expression of the above-mentioned products to stimulate the immune response in mice and its characteristics were initially identified.Results Hantaan virus nucleoprotein and The 26 ku fragment was highly immunogenic in mice, and the antibody titer of immunized mice with ELISA was up to 1:32 0 0 0, and all of them could stimulate different titers (1:10 ~ 1: 16 0) .The passive transfer of splenocytes of immunized mice had a protective effect on Hantaan virus-infected neonatal mice, and the protective effect of spleen cells from 26 ku fragments was more obvious.Conclusion The expression of Hantaan virus nucleoprotein and its amino-terminal The 2 6 ku fragment can stimulate the body to produce neutralizing antibodies, while stimulating cellular immune responses.