摘要
目的：观察丙型肝炎病毒（ＨＣＶ）核心蛋白ＤＮＡ疫苗诱导ＢＡＬＢ／ｃ小鼠体液免疫应答的效力，为研制应用于人类的ＨＣＶＤＮＡ疫苗提供实验依据。方法：将全长ＨＣＶ核心蛋白基因插入真核表达质粒载体ｐｃＤＮＡ３．１，构建ＨＣＶ核心蛋白重组表达质粒ｐｃＤＮＡ３．１ＨＣＶｃｏｒｅ，肌注ＢＡＬＢ／ｃ小鼠，以ＥＬＩＳＡ法检测小鼠血清ＨＣＶ抗体。以此重组质粒转染小鼠ＮＩＨ３Ｔ３细胞，以ＨＣＶ抗体阳性小鼠血清为捕获抗体进行Ｗｅｓｔｅｒｎｂｌｏｔ，检测ＤＮＡ疫苗编码的ＨＣＶ核心蛋白的表达。结果：３０％（３／１０）免疫小鼠产生ＨＣＶ核心蛋白抗体，该抗体能检测到重组质粒在ＮＩＨ３Ｔ３细胞表达的ＨＣＶ核心蛋白。
Objective:To investigate the humoral immune response induced by HCV core DNA vaccine in mice and to furnish evidence for HCV DNA vaccine suitable for human. Methods: HCV core DNA vaccine (pcDNA3.1HCVcore)was constructed by inserting fulllength cDNA of HCV core into an eukaryotic expression vector pcDNA31. BALB/c mice were injected intramuscularly with the recombinant construct and antiHCV core antibody was screened by ELISA. Lysate of NIH3T3 cells transfected with pcDNA31 core was analyzed by Western blot and the antiHCV positive murine sera were used as specific antibodies. Results: ELISA detection revealed that 3 out of 10 vaccinated mice produced HCV antibodies. Western blot analysis showed that the antiHCV core antibodies could detect HCV core protein expressed by NIH3T3 cells. Conclusion: HCV core DNA vaccine is able to induce specific antibodies in mice.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
1998年第3期209-211,共3页
Academic Journal of Second Military Medical University
基金
国家自然科学基金
