恶性疟原虫裂殖子表面蛋白1 DNA与改良痘苗病毒组合疫苗诱导小鼠抗体应答  

DNA/MVA Combined Immunization: Antibody Response to Plasmodium falciparum Merozoite Surface Protein 1 in Mice

作　　者：李淑梅[1] 李珣[2] 薛采芳[1] 缪军[1] 雷俊川[1] 刘忠湘[1] 王宪锋[1] 

机构地区：[1]第四军医大学病原生物学教研室,西安710032 [2]西藏军区总医院,拉萨850003

出　　处：《中国寄生虫学与寄生虫病杂志》2005年第2期93-96,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：联合国开发署/世界银行/世界卫生组织热带病研究与培训特别规划基金课题(A30125)~~

摘　　要：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 DNA/MVA combined immunization in enhancing antibody response to MSP1. Methods DNA vaccine and recombined MVA were constructed based on synthesized MSP1 gene (3D7). BALB/c mice were primed with DNA solely or together with GM-CSF expressing plasmid and then boosted with rMVA/190. Serum IgG and subtype IgG1 and IgG2a were assayed by ELISA. All mice were challenged with allelic replaced Plasmodium berghei. Results Antibodies to MSP1-190 were detected after DNA immunization with an end-point dilution titer of 1:2500. When GM-CSF plasmid was added, the antibody end-point dilution titer reached 1:11150, with an increase of 53 and 10 times respectively after MVA boosting. Among them anti-19 000 antibodies were prominent, 1/4-1/3 of total IgG in serum. However, when the mice were challenged with Pb-PfM19 no prolonged survival was observed(P>0.05). Conclusion High titer antibodies can be elicited in mice by using codon optimized MSP1 gene and DNA/MVA combined immunization. The specificity and protection of these antibodies is being further investigated.

关 键 词：恶性疟原虫裂殖子表面蛋白 抗体应答 痘苗病毒 疫苗诱导 组合 改良 BALB/c小鼠 ELISA测定 DNA免疫 伯氏疟原虫 MSP1 血清IgG MVA CSF 免疫组 人工合成 表达质粒 重组病毒 IgG1 抗体亚类 总IgG 存活时间 MSPI 保护作用 

分 类 号：R392.7[医药卫生—免疫学] R186[医药卫生—基础医学]