B细胞表位基因疫苗诱导特异性免疫应答及特性研究

为研究特异性B细胞表位短肽为基础的DNA疫苗的免疫应答特性,将鸭乙肝病毒Pre-S中的P127-138B细胞表位基因克隆到含多肽表达盒(peptideexpressioncassette,PEC)的载体中,体外转染C2C12肌肉细胞,用DOT-EIA检测表位短肽的表达,进一步分别免疫不同品系的BALB/c和C57BL/6小鼠,比较MHC限制性对免疫应答的影响。结果表明,重组pECk-b1b2载体可在体外瞬时高效表达,并保持原有的免疫活性;体内基因免疫能诱导针对表位短肽的特异性抗体;不同品系小鼠抗体产生动力学相似,但抗体滴度差异显著。这些结果提示,以B细胞表位为基础的基因免疫可诱导特异性免疫应答,为DNA疫苗的分子设计提供了新的思路和应用前景。

Characterization of immune response of DNA vaccine encoding B cell epitope

To explore the immune response of B cell epitope based DNA vaccine, the P127-138 B epitope gene of DHBV Pre-S was cloned into the pECK plasmid constructed in our laboratory. This recombinant plasmid (pECK-b1b2) was transfected into the C2C12 muscle cell line and expression of specific B cell epitope was detected by DOT-EIA. Moreover, the BALB/c (H-2d) and C57BL/6 (H-2b, B6) were immunized by pECK-b1b2 plasmid, respectively. The specific antibody in serum in mice was determined with ELISA. The results showed that B cell epitope of DHBV Pre-S was expressed in the C2C12 cell transfected by pECK-b1b2. Specific antibody against B epitope in sera was also produced after DNA immunization and specific antibody in serum of C57BL/6 was higher than that in BALB/c. These results demonstrated that DNA vaccine based on B epitope could efficiently induce specific immune response with MHC restriction.