人巨细胞病毒gB和pp150融合基因真核表达载体的构建及其免疫活性研究

Objective To provide experimental evidence for development of human cytomegalovirus (HCMV) nucleic acid vaccine,HCMV surface protein(gB),membrane protein(pplSO),and gB-pp150 fused gene eukaryotie expression vector were constructed.Methods gB and pp150 genes were amplified and fused into gB-pp150,then were cloned into pcDNA 3.1(+) to obtain recombinant expression plasmids pcDNA 3.1(+)-gB,pcDNA 3.1(+)-pp150 and pcDNA 3.1(+)-gB-pp150,which were encapsulated with chitosan.Mouse were vaccinated and the humoral and eell immune response were determined by ELISA,specific proliferative response of plenie lymphocytes.Results The gB,pp150 and gB-pp150 fusion gene eukaryotie expression vector were successfully constructed.The antibodies A value induced by peDNA3.1(+)-gB or peDNA3.1(+)-gB-pp150 were much higher than that of peDNA3.1(+)(P<0.01).The IFN-γ levels induced by pcDNA3.1(+)-pp150 and peDNA3.1(+)-gB-pp150 were significantly higher than that of pcDNA3.1(+).There are significant diferenee between the stimulating indexes of pcDNA3.1(+)-pp150 or peDNA3.1(+)-gB-pp150 immunized and normal mice.Conclusion The DNA vaccine pcDNA3.1(+)-gB can induce significant humeral immunity response.and pcDNA3.1 (+)-pp150 can induce high cellular immune response,whereas pcDNA3.1(+)-gB-pp150 can induce both humeral and cellar immune responses in BALB/c mice.

Study on immunogenicity of DNA vaccine encoding human cytomegalovirus gB and pp150 fusion gene