摘要
目的 构建含 HIV－2 gp 105和 gag基因的核酸疫苗，并评价其免疫效果。方法 将 HIV－2 gp 105型和gag 基因克隆到核酸疫苗载体 pcDNA3．l（＋）中，构建重组核酸疫苗质粒。通过间接免疫荧光试验在细胞水平上检测gp105和ggg基因的表达产物。重组核酸疫苗免疫小鼠后，用流式细胞仪测定CD4+、CD8+淋巴细胞亚类数，用HIV－2抗体ELISA检测试剂盒检测免疫鼠血清中抗HIV－2抗体。结果 构建了3种重组核酸疫苗质粒pcgag、pcgp105和pcgag-gp105-gp105，转染BHK细胞后都能表达外源蛋白，免疫小鼠后可有效地刺激淋巴细胞增殖，并诱导产生抗HIV-2特异性抗体，其中pcgag-gp105的免疫效果较pcgag和pcgp105显著。结论 构建的重组核酸疫苗质粒均能诱导机体产生免疫反应，含有融合基因的pcgag-gp105免疫效果最显著。
Abstract: Objective To construct DNA vaccines with HIV - 2 gp105 and gag genes and evaluate the immunological effect of them. Methods A recombinant DNA vaccine plasmid was constructed by cloning HIV - 2 gplOS and gag genes into vector pcDNA 3.1 ( + ). The expressed products of gplOS and gag genes were detected by indirect irnmunofluorescent assay. The CD4+ and CD8+ cells in spleens of mice immunized with the recombinant DNA vaccine were counted using Fluorescence Activated Cell Sorter, and the HIV - 2 antibodies in the sera of them were detected by ELISA kit. Results Three recombinant DNA vaccine plasmids, pcgag, pcgp105 and pcgag- gp105, were constructed. All of them expressed foreign protein after being transformed to BHK cells. The expressed products could stimulate the proliferation of T lymphocytes and induce HIV- 2 specific antibodies in mice. Compared with pcgag and pcgplOS, the immunological effect of pcgag-gplOS was more significant. Conclusion All the constructed recombinant DNA vaccine plasmids can induce immune responses, and that containing both gp105 and gag genes shows more significant effect.
出处
《中国生物制品学杂志》
CAS
CSCD
2002年第1期9-12,共4页
Chinese Journal of Biologicals
基金
国家杰出青年基金项(3977O56661)资助
