细胞与分子免疫学杂志
細胞與分子免疫學雜誌
세포여분자면역학잡지
2001年
1期
52-54
,共3页
黄建生%陈丽珊%解咏梅%张潜%任大明%雷呈祥%沈先荣%贾福星%张丽芸%陈立茵%郭明秋
黃建生%陳麗珊%解詠梅%張潛%任大明%雷呈祥%瀋先榮%賈福星%張麗蕓%陳立茵%郭明鞦
황건생%진려산%해영매%장잠%임대명%뢰정상%침선영%가복성%장려예%진립인%곽명추
丙型肝炎病毒%多表位抗原%基因免疫%免疫应答
丙型肝炎病毒%多錶位抗原%基因免疫%免疫應答
병형간염병독%다표위항원%기인면역%면역응답
目的探讨 HCV复合多表位 DNA疫苗的可行性。方法把 HCV多表位抗原基因 PCX克隆到真核表达载体 pREP9(RSV启动子 )及 pcDNA3 (CMV启动子 )中，构建真核表达载体 pREP9/PCX及 pcDNA3/PCX。将其肌肉注射免疫小鼠及家兔，检测特异性体液免疫和细胞免疫的水平，并观察免疫小鼠的安全性。结果质粒 pREP9/PCX及 pcDNA3/PCX于免疫后第 6wk和第 10 wk时，开始可检测到抗 GZ-PCX IgG，随后抗体滴度逐渐升高，但水平较低，持续时间较短。 pcDNA3/PCX 肌肉注射免疫家兔后，于第 8wk开始出现特异性抗体，至 3个月后滴度升至 1∶ 3 200，随后也开始下降。免疫小鼠及家兔可诱发针对 GZ-PCX融合蛋白的迟发型超敏反应，并刺激淋巴细胞转化。免疫后小鼠的体重正常，肝脾脏未见明显肿大，具有良好的安全性。结论 HCV多表位基因可诱发特异性免疫应答且安全性好，为 HCV疫苗的研究提供了一定的理论及实验依据。
目的探討 HCV複閤多錶位 DNA疫苗的可行性。方法把 HCV多錶位抗原基因 PCX剋隆到真覈錶達載體 pREP9(RSV啟動子 )及 pcDNA3 (CMV啟動子 )中，構建真覈錶達載體 pREP9/PCX及 pcDNA3/PCX。將其肌肉註射免疫小鼠及傢兔，檢測特異性體液免疫和細胞免疫的水平，併觀察免疫小鼠的安全性。結果質粒 pREP9/PCX及 pcDNA3/PCX于免疫後第 6wk和第 10 wk時，開始可檢測到抗 GZ-PCX IgG，隨後抗體滴度逐漸升高，但水平較低，持續時間較短。 pcDNA3/PCX 肌肉註射免疫傢兔後，于第 8wk開始齣現特異性抗體，至 3箇月後滴度升至 1∶ 3 200，隨後也開始下降。免疫小鼠及傢兔可誘髮針對 GZ-PCX融閤蛋白的遲髮型超敏反應，併刺激淋巴細胞轉化。免疫後小鼠的體重正常，肝脾髒未見明顯腫大，具有良好的安全性。結論 HCV多錶位基因可誘髮特異性免疫應答且安全性好，為 HCV疫苗的研究提供瞭一定的理論及實驗依據。
목적탐토 HCV복합다표위 DNA역묘적가행성。방법파 HCV다표위항원기인 PCX극륭도진핵표체재체 pREP9(RSV계동자 )급 pcDNA3 (CMV계동자 )중，구건진핵표체재체 pREP9/PCX급 pcDNA3/PCX。장기기육주사면역소서급가토，검측특이성체액면역화세포면역적수평，병관찰면역소서적안전성。결과질립 pREP9/PCX급 pcDNA3/PCX우면역후제 6wk화제 10 wk시，개시가검측도항 GZ-PCX IgG，수후항체적도축점승고，단수평교저，지속시간교단。 pcDNA3/PCX 기육주사면역가토후，우제 8wk개시출현특이성항체，지 3개월후적도승지 1∶ 3 200，수후야개시하강。면역소서급가토가유발침대 GZ-PCX융합단백적지발형초민반응，병자격림파세포전화。면역후소서적체중정상，간비장미견명현종대，구유량호적안전성。결론 HCV다표위기인가유발특이성면역응답차안전성호，위 HCV역묘적연구제공료일정적이론급실험의거。
Aim To explore the possibility of the multiple epitope DNA vaccines of hepatitis C virus (HCV). Methods A synthetic multiple epitope antigen gene PCX of HCV was cloned into vector pREP9(RSV promoter) and pcDNA3 (CMV promoter) to construct eukaryotic expression vectors pREP9/PCX and pcDNA3/PCX, then they were used to immunize mice and rabbits, the titer of specific humoral and cellular responses were detected and their safety were observed. Results In mice, specific anti-GZ-PCX antibody(IgG) was lower than 1∶ 1 000 and did not persist well. In rabbits, the highest titer of anti-GZ-PCX IgG reached at 1∶ 3 200 and remained for about one month. Delayed type hypersensitivity reactions (DTH)and proliferation response of peripheral lymphocytes were induced by GZ-PCX antigen. Body weights of immunized mice were normal and no obvious toxic reaction was observed. Conclusion The multiple epitope antigen gene of HCV could induce specific immune responses without obvious toxicity and it might be able to serve as an effective HCV vaccine candidate.
