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目的:探讨HBV相关抗原在卡介苗(BCG)中的有效表达及其刺激产生的体液和细胞免疫应答效果。方法:将带有HBV截短C基因和preS1基因片段的穿梭载体转化BCG菌株,筛选重组菌,SDS-PAGE和Western blot实验分析其抗原表达特性;分别以生理盐水、BCG、BCG-pDE22、BCG-pDE22-CS1皮下注射BALB/c小鼠,进行连续抗体测定和细胞毒性T细胞杀伤实验。结果:与对照组相比,重组BCG可表达Mr为24000的新生蛋白,与CS1融合蛋白大小相符,Westernblot实验分析显示出良好的抗原结合特性;带有HBVCS1抗原基因的重组BCG免疫组抗体滴度显著升高,特异性杀伤能力更强。结论:BCG可以作为HBV相关抗原的活载体,为研制HBV新型疫苗提供了实验依据。
Objective: To investigate the effective expression of HBV-related antigens in BCG and the effect of humoral and cellular immune responses induced by their stimuli. Methods: The BCG strain was transformed with the shuttle vector of HBV truncated C gene and preS1 gene, and then the recombinant bacteria were screened by SDS-PAGE and Western blot analysis. The expression of BCG was detected by normal saline, BCG, BCG-pDE22, BCG -pDE22-CS1 BALB / c mice were injected subcutaneously for continuous antibody assays and cytotoxic T-cell killing assays. Results: Compared with the control group, recombinant BCG could express neogenetic protein with Mr 24000, which was consistent with the size of CS1 fusion protein. Western blot analysis showed good antigen binding properties. Antibody titer of recombinant BCG with HBVCS1 antigen gene Significantly increased, specific ability to kill more. CONCLUSION: BCG can serve as a live carrier of HBV-related antigens and provide experimental evidence for the development of new HBV vaccine.