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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。
Objective To construct a nucleic acid vaccine containing gp 105 and gag genes of HIV-2 and evaluate its immunogenicity. Methods HIV-2 gp 105 gene and gag gene were cloned into DNA vaccine vector pcDNA3. l (+), the construction of recombinant nucleic acid vaccine plasmid. Indirect immunofluorescence assay was used to detect the expression products of gp105 and ggg genes at the cellular level. After the mice were immunized with the recombinant DNA vaccine, the subsets of CD4 + and CD8 + lymphocytes were determined by flow cytometry. The anti-HIV-2 antibody in serum of immunized mice was detected by HIV-2 antibody ELISA kit. Results Three recombinant DNA vaccine plasmids pcgag, pcgp105 and pcgag-gp105-gp105 were constructed and transfected into BHK cells to express foreign proteins. After immunization, the mice were stimulated with lymphocyte proliferation and induced anti-HIV-2 Specific antibodies, which pcgag-gp105 immune effect than pcgag and pcgp105 significant. Conclusion The constructed recombinant DNA vaccine plasmid can induce the body to produce immune response, the pcGag-gp105 fusion gene contains the most significant immune effect.