人巨细胞病毒pp65DNA疫苗诱导小鼠免疫效应的研究

目的探讨人巨细胞病毒pp65 DNA疫苗免疫小鼠产生的细胞免疫应答及相关的细胞因子变化,分析该疫苗是否具高度免疫性。方法将40只健康Balb/c小鼠随机分成A、B两组,每组20只。A组(对照组):注射生理盐水;B组(实验组):注射pp65 DNA疫苗;采用流式细胞仪检测CD3+、CD4+及CD8+;采用双抗体夹心ELISA法检测小鼠血清中的IFN-γ;采用放射免疫法检测小鼠血清中的IL-2。统计学处理用SPSS 10.0统计学软件,组间比较用t检验,P0.05为差异有统计学意义。结果B组小鼠T细胞亚群CD4+、CD8+、CD3+及细胞因子IFN-γ,IL-2水平明显高于A组,差异均有统计学意义(P0.01)。结论本研究表明HCMV pp65 DNA疫苗能诱导Balb/c小鼠产生一定的细胞免疫应答及细胞因子的变化。

Study on the mouse immune responses induced by DNA vaccine with the pp65 gene of human cytomegalovirus

Objective To evaluate cellular immune responses induced by the phosphoprotein 65（pp65） DNA vaccine of human cytomegalovirus（HCMV）in Balb/c mice,and to explore the immunity of the DNA vaccine.Methods Forty female Balb/c mice（aged 6-8 weeks）were randomly divided into vaccination（group B,n=20）and control group（group A,n=20）.Mice in group B were injected with 200 μg recombinant pp65 DNA vaccine at 0,2 and 4 weeks respectively,while mice in group A were injected with same volume of saline at the same schedule.CD3＋,CD4＋ and CD8＋ T cells were measured by flow cytometry（FCM）.The method of double antibody sandwich ELISA and radio-immunity assay（RI）were used to detect serum IFN-γ and IL-2 respectively.Results The number of CD3＋,CD4＋ and CD8＋ T cells as well as the concentration of cytokine IL-2 and IFN-γ were significantly higher in group B mice than those in group A mice（P0.01）.Conclusion The pp65 DNA vaccine of HCMV induced specific cellular immune responses and increased the production of cytokines in Balb/c mice.