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目的探索DNA与改良痘苗病毒(MVA)组合免疫对增强恶性疟原虫裂殖子表面蛋白1(MSP1)抗体应答的作用。方法以人工合成MSP1全基因为基础分别构建DNA免疫质粒VR1020/190和重组MVA,单用VR1020/190或与表达质粒GM-CSF共同对小鼠进行初始免疫后,用重组病毒追加强化,采用DNA/MVA组合方案免疫BALB/c小鼠,ELISA测定血清IgG及其亚类水平,经腹腔接种转基因伯氏疟原虫Pb-PfM19进行攻击。结果DNA免疫能有效诱导小鼠产生抗MSP1-190抗体,其终点稀释度为1∶2500,GM-CSF质粒共免疫组抗体的终点稀释度为1∶11150,抗体亚类的测定表明GM-CSF质粒显著促进了IgG1类抗体应答,MVA追加可使单独免疫组和共免疫组抗体分别增加53和10倍;两实验组产生了水平相近的抗19000抗体(1∶32000),其含量占血清中MSP1总IgG的1/4-1/3。经转基因伯氏疟原虫Pb-PfM19攻击后小鼠的存活时间并没有明显延长(P>0.05)。结论采用合成MSP1全基因进行DNA/MVA组合免疫可诱导小鼠产生显著的抗体应答,抗体的详细特性和保护作用正在进一步研究中。
Objective To explore the effect of combined immunization with DNA and modified vaccinia virus (MVA) on enhancing the response of Plasmodium falciparum merozoite surface protein 1 (MSP1) antibody. Methods The DNA immunization plasmids VR1020 / 190 and recombinant MVA were constructed respectively based on the synthetic MSP1 gene. The mice were immunized with VR1020 / 190 alone or with the expression plasmid GM-CSF. BALB / c mice were immunized with the MVA / MVA regimen. Serum IgG and its subclasses were measured by ELISA. The transgenic P. berghei Pb-PfM19 was challenged intraperitoneally. Results DNA immunization can effectively induce mice to produce anti-MSP1-190 antibody with an endpoint dilution of 1: 2500, and the final dilution of the GM-CSF plasmid coimmunization antibody was 1: 11150. The antibody subclass assay indicated that GM-CSF Plasmids significantly promoted the IgG1 antibody response, and MVA supplementation increased antibody levels by 53 and 10 fold, respectively, in both immunized and coimmunized groups; levels of anti-19000 (1: 32,000) MSP1 total IgG 1 / 4-1 / 3. The survival time of mice after the challenge with Pb-PfM19 transgenic berberine was not significantly prolonged (P> 0.05). Conclusion The combination of DNA / MVA combined with MSP1 gene can induce significant antibody response in mice. The detailed characterization and protection of antibody against MSP1 are under further study.