泛素与EB病毒核抗原1融合基因的DNA免疫研究

构建了EB病毒核抗原1(EBNA1)基因的表达质粒pCI-EBNA1和泛素(ubiquitin,Ub)与EBNA1融合基因的表达质粒pCI-Ub-EBNA1.间接免疫荧光和Western blot分析表明,两重组质粒转染HeLa细胞后均能瞬时表达.两种质粒DNA肌肉注射免疫Balb/C小鼠后,分别检测小鼠血清抗EBNA1的抗体和特异性的细胞毒性T淋巴细胞(CTL)反应,比较单基因与融合基因DNA免疫所诱生免疫应答的强度.结果显示:二者诱生抗体的效率无明显差别,但泛素的融合使得针对EBNA1的特异性CTL反应明显增强.

DNA Immunization with Fusion Gene of Ubiquitin and Epstein-Barr Virus Nuclear Antigen 1

Two kinds of expression plasmids were constructed,one of which integrated with EBNA1(pCI-EBNA1),and the other with ubiquitin and EBNA1 fusion gene(pCI-Ub-EBNA1).After transfected into HeLa cell,indirect immunofluorescence assay and Western blot could detect the expression of EBNA1 gene in both plasmids.And then,these plasmids,respectively,were used to intramuscularly immunize Balb/C mice.The serum antibodies and cytotoxic T lymphocyte(CTL)response of the mice were detected.The results showed that there were not much more differences between the two plasmids in antibody response,but the fusion gene DNA induced stronger CTL response than single gene DNA did,which indicated that Ub conjugation did improve the cellular immune response to EBNA1 in mice.