spaA嵌合体核酸疫苗增强抗体水平的研究  

Enhancement of antibodies to protective domain of surface protective antigen A of Erysipelothrix rhusiopathiae by DNA immunization with plasmids expressing spaA-chimeras

作　　者：陈开旭[1] 李轶杰[1] 张富春[1] 曹文尧[1] 李江伟[1] 

机构地区：[1]新疆大学生命科学与技术学院分子生物学重点实验室,新疆生物资源基因工程重点实验室,新疆乌鲁木齐830046

出　　处：《细胞与分子免疫学杂志》2009年第11期984-986,990,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30860265);新疆高校科研计划重点项目(XJEDU2005I05);新疆生物资源基因工程重点实验室基金资助项目(XJDX0201-2005-02)

摘　　要：目的:探讨提高丹毒丝菌表面抗原A(spaA)N端核酸疫苗免疫应答的策略。方法:通过基因重组构建含人α胰岛素抑制剂(AAT)的信号肽、大鼠寡聚软骨基质蛋白(COMP)片段、丹毒丝菌spaA基因N末端及3分子的C3d序列的真核细胞表达质粒pcDNA3-AAT-COMP-spaAN-C3d3(pcD-ACSC)和pcDNA3-spa(pcD-S)。肌注免疫LCR小鼠后,RT-PCR检测小鼠体内丹毒丝菌spaAN基因的瞬时表达,ELISA检测小鼠血清spaA抗体水平的变化,以丹毒丝菌XJ1249评价免疫小鼠的保护效果。结果:免疫第4周时pcD-ACSC核酸疫苗激发了较高水平的SpaAN抗体而pcD-S核酸疫苗激发的抗体水平与对照相比差异不显著;同时pcD-ACSC核酸疫苗具有70%保护效果,pcD-S核酸疫苗则不具保护效果。结论:通过融合高表达分泌蛋白信号肽、增加抗原溶解度的序列和分子佐剂C3d3能显著提高spaAN的抗体水平,为丹毒丝菌spaA核酸疫苗的应用奠定基础。AIM： DNA vaccines expressing protective domain of surface protective antigen A（spaA） of Erysipelothrix rhusiopathiae have been relatively ineffective at generating high-titer, long-lasting, neutralizing antibodies in murine models. METHODS： This paper report using a DNA vaccine expressing a fusion of the spaA protein and various elements, such as a secretion leader sequence from the highly expressed human gene encoding α1-antitrypsin （AAT）, a highly soluble and stably folded domain from the rat cartilage oligomerization matrix protein （COMP）, and three copies of the complement component, C3d3, to enhance the titers of neutralizing spaA-specific antibody. RESULTS： Analysis of titers of the antibody raised in vaccinated mice at different time points indicated that immunizations with the DNA expressing pcDNA3-AAT-COMP-spaAN-3C3d （ （ pcD-ACSC ） ） had higher titers than pcDNA3-spaAN （pcD-S） at weeks 4. Furthermore, the immune protective efficacy of the spaAchimeras was demonstrated by lethal challenge with a virulent homologous strain 1249 against immunized mice. CONCLUSION： These results suggest that using a plasmid vector containing a strong heterologous signal sequence that mediate efficient antigen secretion in vivo and a fused piece of sequence improving antigens solubility, as well as C3d3, genetic adjuvant, could enhance the antibody responses level. This approach might be an efficient way to improve the antibody level of spaAN DNA vaccination.

关 键 词：表面保护抗原A 丹毒丝菌 核酸疫苗 

分 类 号：S852[农业科学—基础兽医学]