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| 1 | DNA immunization with fusion genes encoding different regions of hepatitis C virus E2 fused to the gene for hepatitis B surface antigen elicits immune responses to both HCV and HBV显示文摘AIM: Both Hepatitis B virus (HBV) and Hepatitis C virus(HCV) are major causative agents of transfusion-associatedand community-acquired hepatitis worldwide. Developmentof a HCV vaccine as well as more effective HBV vaccines isan urgent task. DNA immunization provides a promisingapproach to elicit protective humoral and cellular immuneresponses against viral infection. The aim of this study is toachieve immune responses against both HCV and HBV by DNAimmunization with fusion constructs comprising various HCVE2 gene fragments fused to HBsAg gane of HBV.METHODS: C57BL/6 mice were immunized with plasmid DNAexpressing five fragments of HCV E2 fused to the gene forHBsAg respectively. After one primary and one boostingimmunizations, antibodies against HCV E2 and HBsAg weretested and subtyped in ELISA. Splenic cytokine expressionof IFN-γ and IL-10 was analyzed using an RT-PCR assay.Post-immune mouse antisera also were tested for theirability to capture HCV viruses in the serum of a hepatitis Cpatient in vitro.RESUTLTS: After immunization, antibodies against bothHBsAg and HCV E2 were detected in mouse sera, withIgG2a being the dominant immunoglobulin sub-class. High-level expression of INF-γ was deuetected in cultured splenic cells.Mouse antisera against three of the five fusion constructs wereable to capture HCV viruses in an in vitro assay.CONCLUSION: The results indicate that these fusionconstructs could efficiently elicit humoral and Th1 dominantcellular immune responses against both HBV S and HCV E2antigens in DNA-immunized mice. They thus could serve ascandidates for a bivalent vaccine against HBV and HCVinfection. In addition, the capacity of mouse antisera againstthree of the five fusion constnucts to capture HCV virusses invitro suggested that neutralizing epitopes may be present inother regions of E2 besides the hypervariable region 1. | JinJ YangJY 等 | 2002 | World Journal of Gastroenterology2002，8，3： | 18 |
| 2 | 丙型肝炎病毒包膜蛋白E2的DNA免疫显示文摘为了比较启动子及分泌信号对丙型肝炎病毒包膜蛋白E2 (HCVE2 )的DNA免疫效果的影响 ,构建了 4个不同的HCVE2 (384~ 6 6 0 )融合表达质粒 :CMV启动子控制下和EF1α启动子控制下的分泌表达质粒pCMVSec S1E2t6 6 0和pEF1αSec S1E2t6 6 0以及非分泌表达质粒 pCMV S1E2t6 6 0和 pEF1α S1E2t6 6 0。4个表达质粒在HeLa细胞中进行了暂时表达 ,免疫印迹分析表明 ,表达产物都同时具有HBVpreS1及HCVE2蛋白的抗原性 ,夹心ELISA测定结果表明 ,分泌表达质粒的表达量明显高于非分泌表达质粒 ,带CMV启动子的质粒表达量高于EF 1α ;并且只有分泌型质粒转染细胞后 ,才能在细胞培养上清液中检测到融合蛋白。4种表达质粒免疫C5 7BL/ 6小鼠 ,可产生 preS1及E2的抗体 ,比较了抗体转阳率、抗体滴度和维持时间等 ,发现带CMV启动子的E2分泌表达质粒 pCMV S1E2t6 6 0明显优于其他 3组。对 4种质粒产生体液免疫反应不同的可能原因进行了分析。 | 朱珺 王春林 朱立新 孔玉英 汪垣 李光地 | 2002 | 生物化学与生物物理学报2002，34，4： | 1 |
| 3 | 融合有HBV前S1(21~47)片段的HCV E2糖蛋白在CHO细胞中的表达和鉴定(英文)显示文摘在构建丙型肝炎病毒 (HCV)被膜蛋白E2的CHO细胞稳定表达株时 ,为了方便进行表达产物的检测和纯化 ,将编码E2N端 2 77个氨基酸 (384~ 6 6 1)的基因片段和编码乙型肝炎病毒前S1(2 1~ 4 7)区的基因融合 ,一起插入分泌表达载体pSecTagB ,所得重组质粒转染的CHO细胞经Zeocin选择 ,得到稳定细胞株CHO/sS1E2。以E2和 preS1特异的抗体进行免疫印迹分析 ,从筛选到的稳定细胞株检测到了分泌的以及细胞内的表达产物。用糖链类型特异的糖苷酶对表达产物进行了酶解分析 ,发现绝大多数胞内的E2糖蛋白产物是高甘露糖型的 ,而分泌的E2糖蛋白产物则绝大多数是复杂糖化的 ,表明表达的E2在分泌过程中经历了高尔基体的进一步修饰。初步的分析显示 ,融合抗原能特异地和前S1抗体偶联的Sepharose结合 ,并能被顺利地洗脱 ,表明该免疫亲和介质能用于融合抗原的大规模制备。为进一步研究HCVE2糖蛋白的免疫学性质以及重组的HBV/HCV双价疫苗创造了条件。 | 王春林 朱立新 刘晶 张祖传 汪垣 李光地 | 2002 | 生物化学与生物物理学报2002，34，4： | 0 |
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