Wei Qing-kuan, Li Jin, Fu Ting-xia, Bai Xue-lian, Cui Yong, Zhang Dian-bo, Wang Hong-fa, Liu Yu-bing, Fu Bin, Zai De-fu, Huang Bing-cheng, Liu Ke-yi, Han Guang-dong
Shandong Institute of Parasitic Diseases, WHO Collaborating Centre for Lymphatic Filariasis and Taeniasis/ Cysticercosis, Jining 272033, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2006 Oct;24(5):337-41.
To study the protective effect of ROP2 nuclei acid vaccine in mice.
Forty-two BALB/c mice were divided into three groups. Each mouse in experiment group was injected with 50 microg recombinant plasmid pc-DNA3-ROP2 through musculus quadriceps fexoris. In control groups, each mouse was injected with 50 microg blank plasmid pc-DNA3 and with 50 microl PBS respectively. All mice were immunized for three times with an interval of three weeks. The volume was doubled for the final injection in the two plasmid groups. Blood, spleens and lymph nodes of 4 mice in each group were taken for the detection of CD4+, CD8+ T cells and cytokines 2 weeks after the final immunization. The rest mice in 3 groups were challenged with 500 tachyzoites of Toxoplasm gondii RH strain for further observation.
The vaccine induced strong cellular and humoral immune response. The titer of antibody in serum was high after inoculation and recognized ROP2 protein antigen expressed in vitro. The lymphocyte phenotype was analyzed. CD4+ T cells proliferated sharply (69.5+/-3.4)%, and the ratio of CD4/CD8+ increased considerably by (4.69+/-1.32)% (P<0.01). The level of IL-2, IL4, IL-6, IL-12, IFN-gamma and TNF in serum and cultured supernatant of spleen cells and lymph cells was higher in the experiment group than that in control groups, especially in serum. 88.9% mice in the experiment group were protected 180 hours after the challenge of T. gondii. The death time of mice in experiment group was delayed and the survival time was prolonged in comparison to that in control groups with a significant difference (P< 0.01).
The recombinant ROP2 nuclei acid vaccine shows fair immunogeni-city and obviously produces immuno-protection.
研究ROP2核酸疫苗对小鼠的保护作用。
将42只BALB/c小鼠分为三组。实验组每只小鼠通过股四头肌注射50μg重组质粒pc-DNA3-ROP2。对照组每只小鼠分别注射50μg空白质粒pc-DNA3和50μl PBS。所有小鼠均免疫三次,间隔三周。两个质粒组的末次注射量加倍。末次免疫后2周,每组取4只小鼠的血液、脾脏和淋巴结检测CD4⁺、CD8⁺T细胞及细胞因子。其余三组小鼠用500个刚地弓形虫RH株速殖子攻击,进一步观察。
该疫苗诱导了强烈的细胞免疫和体液免疫反应。接种后血清抗体效价高,能识别体外表达的ROP2蛋白抗原。分析淋巴细胞表型,CD4⁺T细胞急剧增殖(69.5±3.4)%,CD4/CD8⁺比值显著增加(4.69±1.32)%(P<0.01)。实验组血清及脾细胞、淋巴细胞培养上清中IL-2、IL4、IL-6、IL-12、IFN-γ和TNF水平均高于对照组,尤其是血清中。实验组88.9%的小鼠在弓形虫攻击后180小时得到保护。与对照组相比,实验组小鼠死亡时间延迟,存活时间延长,差异有统计学意义(P<0.01)。
重组ROP2核酸疫苗具有良好的免疫原性,能明显产生免疫保护作用。