弓形虫DNA疫苗联合诱导BALB/c小鼠保护性免疫力的研究  

Study on the protective immunity induced by the combined use of the surface antigen P30 and P22 DNA vaccine for toxoplasmosis in BALB/c mice

作　　者：占国清[1] 吴少庭[2] 高世同[2] 李国光[3] 

机构地区：[1]郧阳医学院附属人民医院感染性疾病教研室,十堰442000 [2]深圳市疾病预防控制中心 [3]华中科技大学同济医学院流行病与统计学教研室

出　　处：《中国人兽共患病学报》2006年第1期89-91,共3页Chinese Journal of Zoonoses

摘　　要：研究弓形虫表面抗原P30和P22 DNA疫苗联合诱导BALB/c小鼠的保护性免疫作用。48只BALB/c小鼠随机分成4组:A组(NS对照组),肌注生理盐水100μl/鼠;B组(空质粒对照组),肌注pBK-CMV 100μl/鼠(1mg/ml);C组(pBK-P30免疫组)肌注pBK-P30 100μl/鼠(1mg/ml);D组(pBK-P30&pBK-P22联合免疫组)分别肌注pBK-P30和pBK-P22各100μl/鼠(1mg/ml),免疫后5周,通过ConA刺激,MTT法测定免疫鼠脾脏T淋巴细胞转化率,使用间接免疫荧光法,用流式细胞仪对CD4+、CD8+T细胞群进行测定。免疫后10周,免疫鼠腹腔注射弓形虫速殖子攻击感染。结果表明,ConA刺激小鼠脾T淋巴细胞均发生增殖反应,疫苗免疫组略高于两对照组,但4组之间的差异均无显著性(P>0.05)。T淋巴细胞亚群CD4+、CD8+动态分析,CD4+T细胞在各组的增殖均不明显(P>0.05),但两免疫组CD8+T细胞数量升高,CD4+/CD8+比值降低,与空质粒pBK-CMV对照组和NS对照组之间差异均有显著性(P<0.05,P<0.01),pBK-CMV对照组与NS对照组之间差异也有显著性(P<0.01),两免疫组之间差异无显著性(P>0.05)。弓形虫速殖子攻击感染,联合免疫组平均存活时间较两对照组均延长(P<0.05)。但两免疫组间、pBK-P30免疫组与两对照组之间差异无显著性(P>0.05)。弓形虫DNA疫苗能诱导BALB/c鼠产生特异性细胞免疫应答及部分抗虫免疫保护作用,此两种DNA疫苗联合免疫有一定的免疫保护效果。To investigate the protective immunity induced by the combined use of P30 and P22 DNA vaccine for toxoplssmosis, 48 BALB/c mice were randomly divided into 4 groups, in which, in group A （normal saline control group） , each mice was injected with 100μl of normal saline hy intramuscular route; in group B （plasmid pBK-CMV control group）, each mice was injected with 100μl of pBK-CMV（ 1 mg/ml） ; in group C （pBK-P30 group）, each mice was injected with 100μl of pBK-P30 （ 1 mg/ml） and in group D（comhined immunization with pBK-P30 and pBK-P22 group）, each mice was injected with pBK-P30 and pBK-P22（ each mice reeiving 100 pl of pBK-P30 and 100μl of pBK-P22 respectively, 1 mg/ml）. After 5 weeks of immunization, the lymphocyte transformation rate of spleen cells stimulated by Con-A was detected by MTT assay, and the CD^4 ＋ and CD^8 ＋ T cell subgroups were to be assayed by flow cytometery. Ten weeks later, all the injected mice were challenged with the RH strain of T. gondii by intraperitoneal mute. The results showed that the proliferative responses of T lymphocytes of spleen induced by Con-A could be demonstrated in each group of mice, but there was no significant difference among the control and the immunized mice. The kinetic analysis of CD^4＋ and CD^8＋ T cell subgroups showed that the number of CD^4＋ T cells had no obvious increase in all groups, but those of CD^8＋ T cells in two immunized groups showed significant increases with a definite decrease of the CD4/CD8 ratio in comparison with those of pBK-CMV and normal saline controls after 5 weeks of immunization. However, there was remarkable difference between pBKCMV and normal saline controls, but there was no significant differences between group C and group D. The average survival time of group D mice was longer than those of the two control groups of mice, but there were no significant difference between the two immunized groups and among group C and two control groups of mice. It is concluded that the DNA vaccines for toxo

关 键 词：弓形虫 DNA疫苗联合 免疫 

分 类 号：R382[医药卫生—医学寄生虫学]