HIV-2 DNA疫苗免疫小鼠的免疫反应观察

目的：用构建的HIV－2外膜蛋白gp105和核心蛋白gag基因的DNA疫苗免疫小鼠,评价其免疫效果。方法：将HIV－2型gp 105和gag基因克隆到表达载体pIRES1neo中,构建重组DNA疫苗质粒。间接免疫荧光试验证明,构建的DNA疫苗在真核细胞中表达了gp105或／和gag.构建的疫苗免疫小鼠后,用流式细胞仪测定CD4^ 、CD8^ T淋巴细胞亚类数,并用HIV－2抗体ELISA检测试剂盒检测免疫鼠血清中抗HIV－2抗体水平。结果：构建了3种HIV－2 DNA疫苗pIRES1gag、pIRSE1gp105和pIRES1gag-gp105,转染BHK细胞后均能表达抗原蛋白,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pIRES1gag-gp105免疫鼠中,淋巴细胞增殖最显著,而pIRES1gp105免疫鼠中HIV－2特异性抗体水平最高。结论：构建的DNA疫苗均能诱导机体产生免疫反应,其中pIRES1gp105诱导的体液免疫应答最显著,而pIRES1gag-gp105 诱导的细胞免疫响应最显著。

Cellular and humoral immune responses induced by HIV-2 DNA vaccines

Objective To develop HIV 2 DNA vaccines and evaluate their immunological effects in BALB/c mice.Methods HIV 2 env gene and gag gene were inserted into a DNA vaccine expression vector pIRES1neo. The expressed products of env gene and gag gene were examined by indirect immunofluorescent assay (FACS). The CD4 + and CD8 + counts in immunized mice were analyzed via Fluorescence Acti vated cell Sorter and the induces HIV 2 specific antibodies were measured by ELISA Kit for HIV 2.Results Effective expression of HIV 2 antigens was observed in BHK cells transfected with recombinant DNA vaccine plasmids pIRES1gag, pIRES1gp105 and pIRES1gag gp105, the effective proliferation of CD4 + and CD8 + lymphocytes and production of HIV 2 specific antibodies were achieved in immunized mice, the most significant proliferation of CD4 + and CD8 + lymphocytes was achieved in immunized mice by pIRES1gag gp105 and the highest level of HIV 2 specific antibodies was achieved in immunized mice by pIRES1gp105. Conclusion All constructed DNA vaccines can induce both humoral and cellular immunity in immunized mice, pIRES1gag gp105 induces the most remarkable cellular response and pIRES1gp105 induces the most remarkable humoral response in immunized mice.