CAJ | 학술논문

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SpaA_N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.
목적:탐토제고단독사균표면항원A(spaA)N단핵산역묘면역응답적책략.방법:통과기인중조구건함인α이도소억제제(AAT)적신호태、대서과취연골기질단백(COMP)편단、단독사균spaA기인N말단급3분자적C3d서렬적진핵세포표체질립pcDNA3-AAT-COMP-spaAN -C3d3 (pcD-ACSC)화pcDNA3- spa (pcD-S).기주면역LCR소서후, RT-PCR검측소서체내단독사균spaAN기인적순시표체, ELISA검측소서혈청spaA항체수평적변화, 이단독사균XJ1249평개면역소서적보호효과.결과:면역제4주시pcD-ACSC핵산역묘격발료교고수평적SpaA_N항체이pcD-S핵산역묘격발적항체수평여대조상비차이불현저;동시pcD-ACSC핵산역묘구유70%보호효과, pcD-S핵산역묘칙불구보호효과.결론:통과융합고표체분비단백신호태、증가항원용해도적서렬화분자좌제C3d3능현저제고spaAN적항체수평, 위단독사균spaA핵산역묘적응용전정기출.
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-spaA_N(pcD-S) at weeks 4. Furthermore, the immune protective efficacy of the spaA-chimeras 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 spaA_N DNA vaccination.