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重组腺相关病毒载体(rAAV)可在动物体内高水平地持久表达外源基因,本研究采用两种rAAV载体(rAAV1与rAAV2)构建了表达丙型肝炎病毒中国分离株包膜糖蛋白(E1E2)的载体疫苗并以之免疫小鼠,分别采用免疫荧光证实其表达与总抗体,用HCV假病毒系统检测其中和抗体水平,用ELISpot分析其细胞免疫应答,结果表明:rAAV1-E1E2重组载体疫苗单针免疫激发的体液应答明显高于rAAV2-E1E2,rAAV1-E1E2单针注射后3个月可在肌肉组织中检出E2蛋白表达及特异性T细胞应答。上述结果提示HCV重组腺相关病毒载体疫苗单针免疫可引起明显持久的体液与细胞免疫应答。
Recombinant adeno-associated virus vector (rAAV) can persistently express foreign genes in animals at high level. In this study, two rAAV vectors (rAAV1 and rAAV2) were constructed to express the capsid glycoprotein (E1E2) Were used to immunize mice. Immunofluorescence was used to confirm the expression and total antibody respectively. The level of neutralizing antibody was detected by HCV pseudovirus and the cellular immune response was analyzed by ELISpot. The results showed that the recombinant vaccine of rAAV1-E1E2 The body fluid response induced by needle immunization was significantly higher than rAAV2-E1E2. E2 protein expression and specific T cell responses were detected in muscle tissue 3 months after rAAV1-E1E2 injection. The above results suggest that the single-needle immunization of HCV recombinant adeno-associated virus vector vaccine can cause a significant and lasting humoral and cellular immune response.