Yoshida Asuka, Samal Siba K
Virginia-Maryland Regional College of Veterinary Medicine, University of Maryland, College ParkMD, USA.
Front Microbiol. 2017 Apr 20;8:693. doi: 10.3389/fmicb.2017.00693. eCollection 2017.
Avian paramyxovirus serotype 3 (APMV-3) causes infection in a wide variety of avian species, but it does not cause apparent diseases in chickens. On the contrary, APMV-1, also known as Newcastle disease virus (NDV), can cause severe disease in chickens. Currently, natural low virulence strains of NDV are used as live-attenuated vaccines throughout the world. NDV is also being evaluated as a vaccine vector against poultry pathogens. However, due to routine vaccination programs, chickens often possess pre-existing antibodies against NDV, which may cause the chickens to be less sensitive to recombinant NDV vaccines expressing antigens of other avian pathogens. Therefore, it may be possible for an APMV-3 vector vaccine to circumvent this issue. In this study, we determined the optimal insertion site in the genome of APMV-3 for high level expression of a foreign gene. We generated recombinant APMV-3 viruses expressing the green fluorescent protein (GFP) by inserting the GFP gene at five different intergenic regions in the genome. The levels of GFP transcription and translation were evaluated. Interestingly, the levels of GFP transcription and translation did not follow the 3'-to-5' attenuation mechanism of non-segmented, negative-sense RNA viruses. The insertion of GFP gene into the P-M gene junction resulted in higher level of expression of GFP than when the gene was inserted into the upstream N-P gene junction. Unlike NDV, insertion of GFP did not attenuate the growth efficiency of AMPV-3. Thus, APMV-3 could be a more useful vaccine vector for avian pathogens than NDV.
禽副粘病毒3型(APMV-3)可感染多种禽类,但不会在鸡身上引发明显疾病。相反,APMV-1,也被称为新城疫病毒(NDV),可在鸡身上引发严重疾病。目前,天然低毒力的新城疫病毒株在全球被用作减毒活疫苗。新城疫病毒也正在被评估作为一种针对家禽病原体的疫苗载体。然而,由于常规疫苗接种计划,鸡通常预先存在针对新城疫病毒的抗体,这可能导致鸡对表达其他禽病原体抗原的重组新城疫病毒疫苗敏感性降低。因此,APMV-3载体疫苗有可能规避这个问题。在本研究中,我们确定了APMV-3基因组中用于外源基因高水平表达的最佳插入位点。我们通过将绿色荧光蛋白(GFP)基因插入基因组中的五个不同基因间隔区,构建了表达绿色荧光蛋白的重组APMV-3病毒。评估了GFP转录和翻译水平。有趣的是,GFP转录和翻译水平并不遵循非节段性负链RNA病毒的3'到5'衰减机制。将GFP基因插入P-M基因连接处导致GFP的表达水平高于将该基因插入上游N-P基因连接处时的表达水平。与新城疫病毒不同,GFP的插入并未减弱APMV-3的生长效率。因此,与新城疫病毒相比,APMV-3可能是一种更适用于禽病原体的疫苗载体。