摘要
目的探索应用恶性疟原虫DNA疫苗、重组痘苗病毒疫苗和重组蛋白疫苗进行组合免疫接种,诱导针对恶性疟原虫红内期抗原AMA1的保护性抗体。方法PCR扩增云南株恶性疟原虫AMA1编码基因,构建和制备DNA免疫质粒VR1020/E(疫苗D)、重组改良安卡拉痘苗病毒rMVAPE(疫苗V)和重组原核表达AMA1胞外域蛋白E(疫苗P)。用疫苗D单独或与小鼠GM2CSF表达质粒共同对小鼠进行初始免疫,用疫苗V和疫苗P进行单次或先后各一次追加强化。采用疫苗D2V组合方案免疫新西兰白兔。ELISA测定小鼠血清IgG及其亚类IgG1和IgG2a的水平,用免疫动物血清进行疟原虫裂殖子体外入侵抑制实验。结果在疫苗D初始免疫的基础上,采用疫苗V或疫苗P进行强化免疫可显著提高小鼠针对AMA1的抗体应答,小鼠血清特异性IgG平均增加15至137倍,GM2CSF表达质粒对疫苗D2V组合免疫小鼠的抗体应答有显著促进作用。采用疫苗D2V组合免疫在家兔可诱导明显的抗体应答。小鼠和家兔免疫血清在体外可显著抑制疟原虫裂殖子对RBC的入侵。结论将DNA疫苗、重组改良安卡拉痘苗病毒疫苗和重组蛋白疫苗进行组合免疫接种是诱导疟疾红内期保护性抗体的有效方法。
Objective To explore the potential of combined vaccination using DNA vaccine, recombinant vaccinia virus vaccine and recombinant protein vaccine in inducing protective antibodies to malaria antigen AMA1. Methods AMA 1 ectodomain encoding gene was amplified by PCR from the genome of YN strain of Plasmodium falciparum. DNA vaccine plasmid VR1020/E（vaecine D）, recombinant modified vaecinia virus Ankara rMVA/E（vaccine V） and prokaryotic expressed AMA1 ectodomain protein E（vaccine P） were comtructed and prepared. BAIB/c mice were primed two times with vaccine D only or together with GM-CSF expressing plasmid at 0 w and 3 w, at 9 w they were given a booster dose of vaccine V or vaccine P, at 15 w two groups received another booster with different type vaccines. New Zealand rabbits were immunized under vaccine D-V approach. The fiters of IgG in mice sera and the IgG subtypes were determined by EI,ISA. The ability of immune sere in inhibiting plasmodium merozoites invasion were evaluated. Results AMA1 specific antibodies in vaccine D primed mice were greatly enhanced after vaccination with vaccine V or vaccine P, their IgG levels increased by 15 to 137 times. The antibody responses can be promoted in mice immunization by vaccine D-V with additional pcDNA3/GM-CSF. Obvious antibodies were induced in rabbits under vaccine D-V approach. Both mice and rabbit immune sere showed remarkable inhibitory effect on the invasion of merozoites into B.BCs.Conclusion It is a potent way to induce protective antibodies to the intraerythrocytic malaria parasites by combined vaccination of DNA vaccine, recombinant MVA vaccine and recombinant protein vaccine.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2005年第11期949-952,共4页
Chinese Journal of Microbiology and Immunology
基金
军队医药卫生基金课题(01L095)
联合国发展计划/世界银行/世界卫生组织热带病研究与培训特别规划(TDR)项目(A30125)
关键词
恶性疟原虫
疫苗
顶端膜抗原1
改良的安卡拉痘苗病毒
Plasmodium falciparum
Vacine
Apical membrane antigen 1
Modified vaccinia virus Ankara
