摘要
目的 为HCV/HBV重叠或混合感染的防治寻求新的高效联合基因免疫策略。方法 分别将与HCV核心区基因互补的cDNA和HBV核心区基因克隆于真核表达载体 (pRSC) ,构建pRSC HBV/HCV ,转染小鼠骨髓瘤细胞 (SP2 / 0 ) ,通过免疫荧光和Western印迹法检测蛋白的表达 ,免疫Balb/c小鼠 ,用酶联免疫法、乳酸脱氢酶释放法及荷瘤试验观察小鼠体液免疫应答及细胞免疫应答。结果 pRSC HBV/HCV转染的SP2 / 0细胞可见HBcAg及HCV核蛋白染色阳性荧光细胞 ,相对分子质量 14 0 0 0及 2 10 0 0处均可见蛋白条带 ,Western印迹分析可见特异性的蛋白条带。 10只免疫鼠中全部出现抗 HCV及抗 HBV抗体 ,而对照组 (n =10 )全阴性。免疫组小鼠脾细胞对转染pRSC HBV/HCV的SP2 / 0细胞的细胞毒活性明显高于对照组 ,荷瘤试验显示免疫组小鼠注射转染pRSC HBV/HCV的SP2 / 0细胞后肿瘤生长缓慢。结论 pRSC
Objective To develop a HCV combined HBV DNA based therapeutic vaccine. Methods The HBV core gene and HCV core cDNA were inserted into the eukaryotic expression vector with two multiple cloning sites mammalian expression vector, which can be used for expression of two foreign genes, under the CMV promoter and RSC promoter respectively(named pRSC HBV/HCV). Cellular expression of pRSC HBV/HCV was assessed following transfection into SP2/0 cells. The Balb/c mice were immunized by multiple sites intramuscular injection with pRSC HBV/HCV and the immune responses were detected. Results The 21 kd and 14 kd core protein was observed. Both anti HBc IgG and anti HCV core Ab were detected in all immunized mice. Strong CTL activity of splenocytes against SP2/0 cells expressing both HBV and HCV core proteins were measured in immunized mice both in vitro and in vivo. Conclusion The investigation demonstrated that pRSC HBV/HCV could generated both humoral immune response and Strong CTL activity against HBcAg and HCV nucleocapsid in mice.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2002年第2期77-80,共4页
National Medical Journal of China
基金
国家自然科学基金资助项目 (3 9770 660 )
