论文部分内容阅读
目的 :将艾滋病病毒核心蛋白 (gag)与干扰素 (IFNα - 2b)融合基因表达的融合蛋白作为免疫原免疫小鼠 ,动态观察小鼠的体液免疫、细胞免疫与CTL应答。方法 :将IFNα - 2b基因片段插入到gag基因的nt5 31位点 ,经脂质体转染与血凝素阴性蚀斑筛选 ,挑出重组痘苗病毒。经SDS -PAGE和Westernblot鉴定表达产物。以小鼠为实验对象 ,用重组痘苗病毒vJ38gag/IFNα - 2b免疫小鼠 ,用ELISA方法检测血清IgG抗体含量。用流式细胞仪测定小鼠外周血CD+ 4 、CD+ 8T淋巴细胞计数。 3H -TdR掺入法检测细胞毒性T淋巴细胞杀伤活性。结果 :血清IgG抗体含量逐渐增高 ,实验组与对照组比较差异有显著性意义 (p <0 .0 5 )。CD+ 4 、CD+ 8T淋巴细胞计数、CTL检测实验组与对照组比较差异均有显著性意义 (p <0 .0 5 )。结论 :重组痘苗病毒vJ38gag/IFNα - 2b能增强小鼠的体液免疫、细胞免疫和CTL应答。IFNα - 2b可以作为免疫佐剂增强机体的免疫状态。
OBJECTIVE: To immunize mice with the fusion protein of fusion gene of HIV core protein (gag) and interferon (IFNα - 2b) as immunogen to observe the humoral immunity, cellular immunity and CTL response in mice. Methods: IFNα - 2b gene fragment was inserted into nt531 of gag gene. The recombinant plasmid was selected by liposome transfection and hemagglutinin negative plaque screening. The expression products were identified by SDS-PAGE and Western blot. The mice were used as the experimental mice, and the mice were immunized with the recombinant vaccinia virus vJ38gag / IFNα - 2b. The serum IgG was detected by ELISA. Flow cytometry was used to determine the counts of CD + 4 and CD + 8T lymphocytes in peripheral blood of mice. 3H-TdR incorporation assay cytotoxic T lymphocyte killing activity. Results: Serum IgG antibody levels gradually increased, the experimental group compared with the control group, the difference was significant (p <0. 05). CD + 4, CD + 8T lymphocyte count, CTL test in the experimental group compared with the control group were significant differences (p <0.05). Conclusion: Recombinant vaccinia virus vJ38gag / IFNα - 2b can enhance humoral immunity, cellular immunity and CTL response in mice. IFNα - 2b can be used as immune adjuvant to enhance the body ’s immune status.