Mosley Yung-Yi C, Hsieh Ming Kun, Wu Ching Ching, Lin Tsang Long
Department of Comparative Pathobiology, College of Veterinary Medicine, Purdue University, West Lafayette, IN 47907, USA.
Department of Comparative Pathobiology, College of Veterinary Medicine, Purdue University, West Lafayette, IN 47907, USA.
J Virol Methods. 2015 Jan;211:36-42. doi: 10.1016/j.jviromet.2014.10.011. Epub 2014 Oct 28.
DNA vaccine coding for infectious bursal disease virus (IBDV) polyprotein gene and that for avian influenza virus (AIV) hemagglutinin (HA) gene have been shown to induce immunity and provide protection against the respective disease. The present study was carried out to determine whether an IBDV polyprotein gene-based DNA fused with AIV HA gene could trigger immune response to both IBDV and AIV. After transfection, VP2 and HA were detected in the cytoplasm and at cell membrane, respectively, by immunofluorescent antibody double staining method, suggesting the fusion strategy did not affect the location of protein expression. VP4 cleavage between VP2 and HA was confirmed by Western blot, indicating the fusion strategy did not affect VP4 function in transfected cells. After vaccination in chickens, the DNA construct VP24-HA/pcDNA induced ELISA and virus neutralizing antibodies against VP2 and hemagglutination inhibition antibody against the HA subtype. The results indicated that a single plasmid construct carrying IBDV VP243 gene-based DNA fused with AIV HA gene can elicit specific antibody responses to both IBDV and AIV by DNA vaccination.
编码传染性法氏囊病病毒（IBDV）多聚蛋白基因的DNA疫苗以及编码禽流感病毒（AIV）血凝素（HA）基因的DNA疫苗已被证明可诱导免疫并提供针对相应疾病的保护。本研究旨在确定与AIV HA基因融合的基于IBDV多聚蛋白基因的DNA是否能引发对IBDV和AIV的免疫反应。转染后，通过免疫荧光抗体双重染色法分别在细胞质和细胞膜中检测到VP2和HA，这表明融合策略不影响蛋白质表达的位置。通过蛋白质印迹法证实了VP2和HA之间的VP4裂解，表明融合策略不影响转染细胞中VP4的功能。在鸡中接种疫苗后，DNA构建体VP24-HA/pcDNA诱导了针对VP2的ELISA和病毒中和抗体以及针对HA亚型的血凝抑制抗体。结果表明，携带与AIV HA基因融合的基于IBDV VP243基因的DNA的单个质粒构建体可通过DNA疫苗接种引发对IBDV和AIV的特异性抗体反应。
