Li Wen-Shu, Xie Zi-Xin, Chen Qing-Xin, Chen Shao, Zhang Li-Fang
Department of Microbiology and Immunology of Wenzhou Medical College, Wenzhou 325032, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2010 Oct 30;28(5):359-63.
To study the immune response elicited by the recombinant protein vaccine and DNA vaccine of the complex antigen ROP2-SAG1 from Toxoplasma gondii.
Sixty female BALB/c mice were randomly divided into 4 groups (15 per group). Mice in rROP2-SAGI group were immunized subcutaneously with 2.5 microg rROP2-SAG1 protein formulated in Freund's adjuvant. Mice in control group received only adjuvant emulsified with normal saline. Mice in recombinant plasmid pcROP2-SAG1 and control plasmid pcDNA3.1 groups were each injected intramuscularly with 100 microg of pcROP2-SAG1 and pcDNA3.1, respectively. All the mice received three immunizations at 2-week intervals. Serum samples were collected at 25, 45, and 70 days after immunization for determining antibody IgG, and at 2 weeks after the last immunization IgG1 and IgG2a were detected all by ELISA. Cell counting kit-8 (CCK-8) was used to determine the splenocyte proliferation, and the supernatant of cultured splenocytes was collected for the detection of IFN-gamma by ELISA.
The level of IgG continued to rise in rROP2-SAG1 group after immunization, and similarly in pcROP2-SAG1 group. At 2 weeks after the last immunization, level of IgG1 (1.538 +/- 0.183) was higher than that of IgG2a (0.618 +/- 0.122) (P < 0.05) in rROP2-SAG1 group. Whereas no significant difference between IgG1 (1.107 +/- 0.137) and IgG2a (0.830 +/- 0.185) was observed in pcROP2-SAG1 group (P > 0.05). Compared with the pcROP2-SAG1 group (A450 = 0.123 +/- 0.018), more significant proliferation response of splenocytes was observed in rROP2-SAG1 group (0.348 +/- 0.042) (P < 0.05). There was no significant difference (P > 0.05) of IFN-gamma and IL-2 in the supematant of cultured splenocytes between the groups of rROP2-SAG1 and pcROP2-SAG1.
The antibody level and splenocyte proliferation have been significantly higher in mice immunized with recombinant protein rROP2-SAG1 than those with recombinant plasmid pcROP2-SAG1.
研究弓形虫复合抗原ROP2-SAG1重组蛋白疫苗和DNA疫苗引发的免疫反应。
将60只雌性BALB/c小鼠随机分为4组(每组15只)。rROP2-SAGI组小鼠皮下注射用弗氏佐剂配制的2.5μg rROP2-SAG1蛋白。对照组小鼠仅接受用生理盐水乳化的佐剂。重组质粒pcROP2-SAG1组和对照质粒pcDNA3.1组小鼠分别肌肉注射100μg的pcROP2-SAG1和pcDNA3.1。所有小鼠每隔2周进行3次免疫。在免疫后25、45和70天采集血清样本以测定抗体IgG,在最后一次免疫后2周通过ELISA检测IgG1和IgG2a。使用细胞计数试剂盒-8(CCK-8)测定脾细胞增殖,并收集培养脾细胞的上清液通过ELISA检测IFN-γ。
免疫后rROP2-SAG1组IgG水平持续升高,pcROP2-SAG1组情况类似。在最后一次免疫后2周,rROP2-SAG1组IgG1水平(1.538±0.183)高于IgG2a水平(0.618±0.122)(P<0.05)。而pcROP2-SAG1组IgG1(1.107±0.137)和IgG2a(0.830±0.185)之间未观察到显著差异(P>0.05)。与pcROP2-SAG1组(A450 = 0.123±0.018)相比,rROP2-SAG1组脾细胞增殖反应更显著(0.348±0.042)(P<0.05)。rROP2-SAG1组和pcROP2-SAG1组培养脾细胞上清液中IFN-γ和IL-2无显著差异(P>0.05)。
用重组蛋白rROP2-SAG1免疫的小鼠抗体水平和脾细胞增殖显著高于用重组质粒pcROP2-SAG1免疫的小鼠。