CAJ | 학술논문

目的构建含有SIVgag基因的DNA疫苗和重组腺病毒疫苗,为后期在SIV感染的猴模型中进行多载体疫苗联合免疫策略的治疗效果评价奠定基础.方法将SIV gag基因按照哺乳动物偏嗜密码子进行优化并构建至pVR载体,作为DNA疫苗.以Western Blot方法比较优化前后gag基因表达水平.将优化后的gag基因插入重组腺病毒载体,构建rAd5-SIVgag疫苗.在BALB/c小鼠中分别比较DNA疫苗及rAd5-SIV.gag疫苗单独及联合免疫的效果.结果密码子优化的SIVgag基因的表达水平远高于野生型SIVgag基因.重组腺病毒疫苗免疫一次或两次诱导的细胞免疫反水平分别高于DNA疫苗免疫一次或两次.两种疫苗联合免疫的反应水平与腺病毒疫苗免疫两次的水平相当,高于DNA疫苗单独免疫及腺病毒疫苗单独免疫一次的结果.结论成功优化了SIV gag基因,使其不依赖Rev高水平表达；成功构建了表达优化后SIV gag基因的DNA疫苗和rAd5疫苗,可以在小鼠体内诱导较强的gag基因特异性CTL应答.
목적 구건함유SIVgag기인적DNA역묘화중조선병독역묘,위후기재SIV감염적후모형중진행다재체역묘연합면역책략적치료효과평개전정기출.방법 장SIV gag기인안조포유동물편기밀마자진행우화병구건지pVR재체,작위DNA역묘.이Western Blot방법비교우화전후gag기인표체수평.장우화후적gag기인삽입중조선병독재체,구건rAd5-SIVgag역묘.재BALB/c소서중분별비교DNA역묘급rAd5-SIV.gag역묘단독급연합면역적효과.결과 밀마자우화적SIVgag기인적표체수평원고우야생형SIVgag기인.중조선병독역묘면역일차혹량차유도적세포면역반수평분별고우DNA역묘면역일차혹량차.량충역묘연합면역적반응수평여선병독역묘면역량차적수평상당,고우DNA역묘단독면역급선병독역묘단독면역일차적결과.결론 성공우화료SIV gag기인,사기불의뢰Rev고수평표체；성공구건료표체우화후SIV gag기인적DNA역묘화rAd5역묘,가이재소서체내유도교강적gag기인특이성CTL응답.
Objective To construct DNA and recombinant adenovirus vector vaccines containing SIV gag gene for future evaluation of therapeutic effects of combined immunization strategy in SIV-infected macaque models.Methods The modified SIV gag gene was cloned into pVR vector to get DNA vaccine.The wild type and codon-modified SIV gag gene expression was analyzed by Western Blot assay.Then rAdSSIVgag vaccine expressing modified SIV gag was constructed using AdmaxTM system.BALB/c mice were immunized with DNA or rAd5 vaccine once or twice alone or in DNA prime/rAd5 boost strategy.Gag specific cellular immune responses were detected by IFN-γ ELISPOT assay two weeks post the last immunization.Results The codon modification increased the expression of Gag protein significantly.The cellular immune responses in mice immunized with rAd5 vaccine alone once or twice were higher than that of DNA vaccine.And the immune responses elicited by DNA prime/rAd5 boost were comparative with that of immunization with rAd5 twice.Conclusion The codon modification of SIV gag gene was successful.DNA and rAd5 vaccines encoding codon-modified SIV gag were constructed and both can induce strong Gag-specific CTL responses in mice.