CAJ | 학술논문

目的构建人巨细胞病毒(human cytomegalovirus,HCMV)糖蛋白(gB)和胞膜蛋白(pp150)的单基因及融合基因的真核表达载体并比较其免疫学活性,为研制HCMV核酸疫苗奠定实验基础.方法 PCR扩增HCMV gB及pp150并将其连接成融合基因,分别克隆至peDNA3.1(+)真核表达载体;制备纳米粒DNA疫苗并免疫BALB/e小鼠,检测其体液免疫与细胞免疫应答.结果成功制备了peDNA3.1(+)-gB、peDNA3.1(+)-pp150和peDNA3.1(+)-gB-pp150纳米核酸疫苗;gB和gB-pp150核酸疫苗均能刺激小鼠产生较强的特异性抗体(P<0.01);pp150和gB-pp150核酸疫苗免疫组的小鼠脾细胞能产生较多的IFN-γ和较高的刺激指数.结论 HCMV gB单基因核酸疫苗能诱导较强的体液免疫应答,pp150单基因核酸疫苗则能诱导较强的细胞免疫应答;而gB-pp150的融合基因疫苗能诱导较强的体液免疫和细胞免疫应答.
목적 구건인거세포병독(human cytomegalovirus,HCMV)당단백(gB)화포막단백(pp150)적단기인급융합기인적진핵표체재체병비교기면역학활성,위연제HCMV핵산역묘전정실험기출.방법 PCR확증HCMV gB급pp150병장기련접성융합기인,분별극륭지peDNA3.1(+)진핵표체재체;제비납미립DNA역묘병면역BALB/e소서,검측기체액면역여세포면역응답.결과 성공제비료peDNA3.1(+)-gB、peDNA3.1(+)-pp150화peDNA3.1(+)-gB-pp150납미핵산역묘;gB화gB-pp150핵산역묘균능자격소서산생교강적특이성항체(P<0.01);pp150화gB-pp150핵산역묘면역조적소서비세포능산생교다적IFN-γ화교고적자격지수.결론 HCMV gB단기인핵산역묘능유도교강적체액면역응답,pp150단기인핵산역묘칙능유도교강적세포면역응답;이gB-pp150적융합기인역묘능유도교강적체액면역화세포면역응답.
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.