中国寄生虫学与寄生虫病杂志 ›› 2005, Vol. 23 ›› Issue (2): 8-96.

• 论著 • 上一篇    下一篇

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

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   

  1. 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
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2005-04-30 发布日期:2005-04-30
  • 通讯作者: 薛采芳

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

LI Shu-mei;LI Xun;XUE Cai-fang;MIAO Jun;LEI Jun-chuan;
LIU Zhong-xiang;WANG Xian-feng
  

  1. Department of Etiology, The Fourth Military Medical University, Xi′an 710032, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2005-04-30 Published:2005-04-30
  • Contact: XUE Cai-fang

摘要: 目的?摇探索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∶2 500,GM-CSF质粒共免疫组抗体的终点稀释度为1∶11 150,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19 000抗体(1∶32 000),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。

关键词: 裂殖子表面蛋白1, 疫苗, DNA, 改良痘苗病毒, 恶性疟原虫

Abstract: 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.

Key words: Merozoite surface protein 1, Vaccines, DNA, Modified Vaccinia virus Ankara, Plasmodium falciparum