汉滩病毒部分核蛋白基因原核表达产物的免疫及其保护作用

Study on the immunogenicity and immune protection of prokaryotic expressed products of partial the gene of Hantaan virus nucleocapsid protein (HTNV NP)

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 study the immunogenicity and immune protection of prokaryotic expressed products of partial gene (37 294bp) of HTNV NP (NP aa1 86). Methods BALB/c mice were immunized with purified recombinant NP aa1 86 peptide. The antibodies against HTNV were detected by IFA, lymphocytes blasto genesis function and cytotoxin T lymphocytes (CTL) effect of the immunized mice were detected by 3H TdR labelled lymphocyte transformation and 51 Cr released from target cells within 4 hours, and immune protection were studied by passively transferring splenic cells from immunized mice into infant mice with fatal HTNV infection. Results The antibodies against HTNV were detected in the sera of mice 1 week after immunization and its titer was up to 1∶12?800 after 4 weeks. Both stimulation index (SI) of lymphocytes blasto genesis and killing rate of CTL in the immunized mice were high than that of the controls ( P <0.01), but no significant difference was found between the immunized mice and mice immunized by recombinant full length HTNV NP ( P >0.05). The transferred immunized splenic cells could partially protect the infant mice from HTNV fatal infection (protective rate was about 25%). Conclusion The recombinant NP aa1 86 peptide contains almost all humoral immune epitopes and partial cellular immune epitopes which can elicited certain immune protection against HTNV infection.