丙型肝炎病毒及恶性疟原虫复合多表位抗原基因与谷胱甘肽转硫酶基因的融合表达及其在小鼠中免疫应答的研究

目的：探讨复合多表位丙型肝炎病毒（ＨｅｐａｔｉｔｉｓＣｖｉｒｕｓ，ＨＣＶ）及恶性疟原虫（Ｐｌａｓｍｏｄｉｕｍｆａｌｃｉｐａｒｕｍ，Ｐｆ）双价疫苗的可行性。方法：将人工合成的复合多表位ＨＣＶ基因ＰＣＸ及Ｐｆ抗原基因ＡＢ（ＳＰｆ６６＋ＳＰｆ１０５）克隆到谷胱甘肽转硫酶（ＧｌｕｔａｔｈｉｏｎｅＳｔｒａｎｓｆｅｒａｓｅ，ＧＳＴ）表达载体ｐＧＥＸ２Ｔ中，并在大肠杆菌中高效表达ＧＳＴＨＣＶＰｆ复合抗原（ＧＳＴＣＡＢ），分析表达产物的免疫特异性、免疫原性及安全性。结果：ＧＳＴＣＡＢ可被ＨＣＶ患者血清、鼠抗ＧＺＰＣＸ及鼠抗Ｐｆ抗体特异识别，可诱发小鼠产生针对ＧＳＴＣＡＢ、ＧＺＰＣＸ及ＰｆＡｇ的特异性体液免疫应答及针对ＧＳＴＣＡＢ、ＧＺＰＣＸ的迟发性超敏反应，免疫后ＣＤ８＋Ｔ细胞比例略为升高，免疫小鼠未见明显的毒副作用。结论：ＧＳＴＣＡＢ融合蛋白具有良好的ＨＣＶ及Ｐｆ免疫特异性，可诱发理想的体液及细胞免疫应答，可望成为ＨＣＶＰｆ双价疫苗的候选抗原。

Expression of multi-epitopes antigen gene of hepatitis C virus and plasmodium falciparum fused to glutathione S-transferase gene and studies on its immunogenicity in mice

Objective: To explore the possibility of bivalent vaccine corresponding to hepatitis C virus (HCV) and plasmodium falciparum (Pf). Methods: Two synthetic multiple epitopes antigen genes PCX of HCV and AB of Pf were cloned into glutathione Stransferase fusion expression vector pGEX2T to highly express the fusion antigen GSTCAB. The immunological specificity,immune response and safety in mice were studied. Results: GSTCAB specifically reacted with antiHCV and antiPf sera, and induced highly specific antibodies, which could recognize GSTCABGZPCX antigen and the ratio of CD8 T cells was slightly higher as compared with that of blank control. Conclusion: The results show the hybrid antigen GSTCAB has excellent antigenicity and might be able to serve as a bivalent vaccine candidate specific for both hepatitis C virus and plasmodium falciparum.