关键词: 人, 疫苗, DNA, 糖蛋白B截短基因, 流式细胞术, 羧基荧光素二醋酸盐琥珀酰亚胺酯

Abstract: To construct an eukaryotic expression plasmid containing the truncated gene encoding the part of herpes simplex virus type 1 glycoprotein B(HSV-1,gB) and evaluate its effects on cell-mediated immunity.Methods The part encoding sequence of the glycoprotein B 14-507 amino acid was amplified from HSV-1 SM44 DNA genome by polymerase chain reaction (PCR)，and then was directionally cloned into eukaryotic expression vector pcDNA₃，the recombinant vector pcDNA₃-gBt was confirmed by the restrictived endonuclease analysis，PCR and sequence analysis.It was employed to evaluate immune response of the mice inoculated triply with the DNA vaccine.The transformation of CD4⁺/CD8⁺T lymp hocyte was detected by means of antibody and the influence of cell-mediated immunity represented by CTL was assayed through CFSE/PI flow cytometry.Results The construction of the HSV-1 gBt DNA vaccine was identified; cytotoxic activity was strengthened; the amount of CD4⁺ T cell from BALB/c mice immuned with pcDNA₃-gBt was higher than mice immuned with plasmid pcDNA₃and saline，the tendacy was the same for CTL，but the amount of CD4⁺ T cell and CD4⁺/CD8⁺ from BALB/c mice immuned with pcDNA₃-gBt had no obvious difference with mice immuned with plasmid pcDNA₃ and saline.Conclusion The HSV-1 gBt DNA vaccine is powerful to induce cell-mediated immunity .

Key words: human, vaccines, DNA, glycoprotein B truncated gene, flow cytometry, carbox fluorescenceindiacetate succinimidyl ester