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目的 探讨汉滩病毒 (HTNV)部分核蛋白 (NP)基因 (37～ 2 94bp)原核表达产物NPAA1～ 86多肽的免疫原性及其免疫保护作用。方法 大肠杆菌表达的HTNV部分核蛋白多肽NPAA1～ 86混合福氏佐剂 ,腹腔注射免疫BALB/c小鼠 ,IFA检测抗体应答 ;3H TdR掺入及 4h51Cr释放试验检测免疫小鼠脾细胞对HTNV的特异淋巴细胞增殖转化指数及细胞毒T淋巴细胞 (CTL)杀伤功能 ;免疫小鼠脾细胞转输感染HTNVA9株的乳鼠 ,观察对乳鼠致死性的免疫保护作用。结果 免疫后 1周小鼠即出现抗体应答反应 ,抗体滴度最高达 1∶12 80 0 ,与重组完整NP免疫组差异无显著性 (P >0 0 5 ) ;免疫小鼠的淋巴细胞增殖转化指数、CTL杀伤率较对照组明显增高 (P <0 0 1) ,与完整NP免疫组差异无显著性 (P >0 0 5 ) ,CTL杀伤率随效∶靶比例的增高呈逐渐上升趋势 ;免疫小鼠脾细胞转输可部分保护病毒致死性感染的乳鼠 ,保护率约 2 5 %。结论 大肠杆菌表达的汉滩病毒部分核蛋白多肽NPAA1～86不仅包含NP几乎所有的体液免疫表位 ,同时含有部分细胞免疫表位 ,对汉滩病毒感染可产生部分免疫保护作用。
Objective To investigate the immunogenicity and immunogenicity of prokaryotic expression product NPAA1 ~ 86 of partial nuclear protein (NP) gene of Hantaan virus (HTNV) (37 ~ 2 94bp). Methods BALB / c mice were immunized with NPAA1 ~ 86 mixed with Freund’s adjuvant (HTAA), and IFA was used to detect the antibody response. 3H TdR incorporation and 4h51Cr release assay were used to detect the effect of immunized mouse spleen cells on HTNV Specific lymphocyte proliferation and transformation index and cytotoxic T lymphocyte (CTL) killing function. The spleen cells of immunized mice were transfused to HTNVA9 strain of suckling mice to observe the immune protective effect on the lethality of suckling mice. Results One week after immunization, the antibody response appeared in mice with antibody titers up to 1:12 80 0, which showed no significant difference from that of the recombinant complete NP (P> 0.05). The proliferation of lymphocytes in immunized mice (P <0.01). There was no significant difference between CTL and whole NP group (P> 0.05). The killing rate of CTL increased gradually with the increase of target: Immune mouse spleen cell transfusion can be part of the protection of the virus lethal infection in neonatal rats, the protection rate of about 25%. Conclusion Some Hantaan virus nucleoprotein NPAA1 ~ 86 expressed in Escherichia coli not only contain almost all the humoral immune epitopes of NP, but also contain some cellular immunophenotypes, which may partially protect the Hantaan virus from infection.