CAJ | 학술논문

本文旨在分析体内电穿孔（EP）技术对 DNA载体 pDRVI1．0表达效率和人类免疫缺陷病毒1型（HIV-1）DNA疫苗免疫反应的辅助效果，为其在DNA疫苗中的应用提供参考数据。通过构建携带荧光素酶基因的pDRVI1．0-Fluc质粒，利用活体成像技术分析EP接种对荧光素酶蛋白的组织分布、表达水平和持续时间的影响；同时，构建携带我国HIV-1 CRF07＿BC流行毒株 env基因的DNA疫苗pDRVI1．0-HIV ，利用酶联免疫斑点法（ELISPOT）、酶联免疫吸附试验（ELISA）和中和抗体法对 EP辅助免疫反应的特点进行分析。结果显示，EP接种后，pDRVI1．0-Fluc质粒未改变组织分布特点，但其体内表达效率显著提高，载体的饱和接种量降低。同时，EP技术提高了pDRVI1．0-HIV疫苗免疫小鼠后诱导的γ干扰素（IFN-γ）分泌型T细胞反应和Env特异性结合抗体效价。结果提示，EP技术可在DNA疫苗应用方面发挥作用。
본문지재분석체내전천공（EP）기술대 DNA재체 pDRVI1．0표체효솔화인류면역결함병독1형（HIV-1）DNA역묘면역반응적보조효과，위기재DNA역묘중적응용제공삼고수거。통과구건휴대형광소매기인적pDRVI1．0-Fluc질립，이용활체성상기술분석EP접충대형광소매단백적조직분포、표체수평화지속시간적영향；동시，구건휴대아국HIV-1 CRF07＿BC류행독주 env기인적DNA역묘pDRVI1．0-HIV ，이용매련면역반점법（ELISPOT）、매련면역흡부시험（ELISA）화중화항체법대 EP보조면역반응적특점진행분석。결과현시，EP접충후，pDRVI1．0-Fluc질립미개변조직분포특점，단기체내표체효솔현저제고，재체적포화접충량강저。동시，EP기술제고료pDRVI1．0-HIV역묘면역소서후유도적γ간우소（IFN-γ）분비형T세포반응화Env특이성결합항체효개。결과제시，EP기술가재DNA역묘응용방면발휘작용。
This study aims to evaluate the effect of electroporation on the expression of DNA vaccines in vivo and induced adaptive immune responses .The gene expression in vivo following DNA delivery via electropo-ration was determined by assessing reporter gene products of firefly luciferase by using in vivo imaging de-vice ,which was further dissected as the tissue distribution of gene products ,the peak level and the lasting time of expression when compared to the traditional immunization through muscular injection . The data showed that electroporation did not alter the tissue distribution ,but increased the peak level and lowered the plateau dosage of DNA vaccine .Further experiments with DNA vaccine pDRVI1-HIV expressing env from human immunodeficiency virus type 1 (HIV-1) CRF07_BC demonstrated that electroporation was able to enhance cellular and humoral immune responses which were gauged by the level of interferon-γ enzyme-linked immunospot (ELISPOT ) responses and HIV-1 specific binding antibody titers , respectively . Altogether ,electroporation in vivo technology is likely to be a useful tool in the administration of DNA vaccines .