汉滩病毒囊膜糖蛋白G1、G2腺病毒载体的表达及免疫分析

为了研究利用腺病毒载体表达汉滩病毒囊膜糖蛋白G1、G2的可行性及免疫原性。通过克隆76-118株G1、G2基因至腺病毒表达载体pAdTrackCMV，得到阳性克隆padTrackCMV-G1、G2。PmeI线性化的阳性克隆与腺病毒骨架载体pAdeasy-1共转化BJ5183宿主菌，经同源重组后得到重组病毒rAdeasy-G1、rAdeasy-G2。重组病毒经PacI线性化后，脂质体介导转染293细胞，使重组病毒得到扩增。将重组病毒免疫Balb／c小鼠，并通过ELISA和间接免疫荧光对免疫小鼠血清进行了分析。结果表明，rAdeasy—G1组六只免疫小鼠、rAdeasy—G2组4只免疫小鼠均产生了能与汉滩病毒抗原发生反应的特异抗体。该研究为进一步研制以腺病毒为活载体的汉坦病毒工程疫苗奠定了基础。

Expression and Immunologenic Study of the Hantaan Virus Glycoprotein G1 and G2 by Using the Adenoviral Vector

In order to study the immune responses of Hantaan virus envelope glycoproteins G1 and G2 in adenovirus expression system, G1 and G2 gene were amplified from Hantaan virus 76-118 and cloned to the plasmid pAdTrackCMV. Positive clones pAdTrackCMV-G1 and G2 were determined with restriction enzyme digestion. The recombinant adenovirus rAdeasy-G1 and rAdeasy-G2 were generated by bacterial homologous recombination with adenoviral plasmid pAdeasy-1 in E.coli BJ5183. After amplification in the 293 cell, the recombinant viruses were immuned to the Balb/c mice. The antibody reaction were detected by ELISA and Immunofluorescence Assay. Six rAdeasy-G1 immuned mice and four rAdeasy-G2 immuned mice produce antibody which can specifically react with the Hantaan virus. These results suggested a possible application of adenovir- us expression system.