论文部分内容阅读
目的将艾滋病病毒外膜蛋白(env)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,观察小鼠的免疫功能状态和细胞免疫应答。方法将IFNα-2b基因片段插入到env基因的下游,经脂质体转染,筛选重组痘苗病毒,经SDS-PAGE和Western blot鉴定表达产物。用重组病毒vJ16env/IFNα-2b免疫小鼠,以生理盐水和野生型痘苗病毒作为对照,检测小鼠脾淋巴细胞对ConA、LPS及IgG的反应性;用流式细胞仪测定小鼠脾淋巴细胞CD4、CD8T细胞计数与CTL。结果与对照组比较,实验组脾淋巴细胞对ConA、LPS及IgG的反应性显著增高(P<0.05);CD4T淋巴细胞计数和CTL活性也显著增高(P<0.05);CD8T淋巴细胞计数呈增高趋势,但未达到显著意义的程度。结论重组痘苗病毒vJ16env/IFNα-2b能增强小鼠的免疫功能和诱导细胞免疫。
Objective To immunize mice with the fusion protein expressed by the fusion protein of HIV-1 and interferon (IFNα-2b), and to observe the immunological status and cellular immune response in mice. Methods The IFNα-2b gene fragment was inserted into the downstream of env gene. The recombinant vaccinia virus was screened by lipofectamine 2000 and identified by SDS-PAGE and Western blot. The mice were immunized with the recombinant virus vJ16env / IFNα-2b, and the reactivity of splenic lymphocytes to ConA, LPS and IgG was detected by using normal saline and wild type vaccinia virus. The splenic lymphocytes CD4, CD8 T cell count and CTL. Results Compared with the control group, the reactivity of splenic lymphocytes to ConA, LPS and IgG in experimental group was significantly increased (P <0.05), the counts of CD4 T lymphocytes and the activity of CTL were also significantly increased (P <0.05), and the counts of CD8 T lymphocytes were increased Trend, but not to the point of significance. Conclusion Recombinant vaccinia virus vJ16env / IFNα-2b can enhance immune function and induce cellular immunity in mice.