摘要
目的用pcDNA_3－P30真核表达质粒直接免疫小鼠，观察所诱导的小鼠细胞免疫应答。方法大量制备量级质粒pcDNA_3－P30，经肌肉注射BALB／c小鼠，每隔3周接种1次，一共免疫3次。用MTT方法对脾脏的NK杀伤细胞率和淋巴细胞的转化率进行测定，采用免疫荧光法对CD4+、CD8+细胞进行测定。结果实验组NK细胞杀伤率为：70．0％±3．64，对照组及空白对照组分别为48．5％±6．06和470％±5．93，实验组NK细胞活性比对照组明显增高（P＜0．05）；ConA刺激小鼠淋巴细胞转化实验，实验组与对照组及空白对用级差异无显著性（P＞0．05）;对T淋巴细胞亚群CD4＋、CD8＋进行动态分析，可见随着感染时间的延长，CD8＋的数量逐渐上升，CD4＋／CD8＋的比率逐渐下降，实验组与对照组及空白对照有明显差异（P＜0．05）。结论重组质粒pcDNA3-p30免疫BALB／c小鼠可诱导一定的细胞免疫。
Aim Directly immunize mice with the recombinant plasmid pcDNA_3-p30,to observe the cellular immuneresponses induced by the DNA vaccine. Methods Large scale prepared recombinant plasmid pcDNA_3-p30 was injected intothe BALB/c mice via muscles. Two booster injections were given at the 3rd week and 6th week followed the first injection.The killing activity of NK cells and the proliferation rate of spleen cells were tested by MTT assay,and T cell subgroupCD4+ 、CD8+ were tested by the indirect immune fluorescent assay. Result The killing rate of the NK cells from immunizedmice was 70. 0% ± 3. 64,and that of control groups were 48. 5 % ±6. 06 and 47.0% ±5. 93 ;The killing activity of NK cells inthe immunized mice was remarkable higher than that of control groups(P＜0.05) ;Cthe results of kinetic analysis of CD4+,CD8+ T cell subgroup showed that the number of CD8+ gradually increased and the rate of CD4+/CD8+ gradually reducedwith time prolong. There was remarkable differenc between control mice and immunized ones (P＜0. 05) ;the proliferating re-sponses of control group and immunized mouse spleen cells were generated when stimulated by ConA,but there is no remark-able difference between control mice and immunized ones (P＞0. 05).Conclusion pcDNA_3-p30 vaccine could elicit cellular im-mune responses.
出处
《中国人兽共患病杂志》
CSCD
北大核心
1999年第3期11-13,共3页
Chinese Journal of Zoonoses
基金
中山医科大学211工程重点学科建设课题
关键词
弓形虫
DNA疫苗
P30基因
细胞免疫应答
Toxoplasma gondii
Recombinant plasmid pcDNA_3-p30
DNA vaccine
Cellular immune
