Virginia-Maryland Regional College of Veterinary Medicine, University of Maryland, College Park, MD 20740, USA.
Department of Poultry Diseases, Faculty of Veterinary Medicine, Cairo University, Giza 12211, Egypt.
Viruses. 2021 Feb 19;13(2):316. doi: 10.3390/v13020316.
A reverse genetic system for avian paramyxovirus type-3 (APMV-3) strain Wisconsin was created and the infectious virus was recovered from a plasmid-based viral antigenomic cDNA. Green fluorescent protein (GFP) gene was cloned into the recombinant APMV-3 genome as a foreign gene. Stable expression of GFP by the recovered virus was confirmed for at least 10 consecutive passages. APMV-3 strain Wisconsin was evaluated against APMV-3 strain Netherlands and APMV-1 strain LaSota as a vaccine vector. The three viral vectors expressing GFP as a foreign protein were compared for level of GFP expression level, growth rate in chicken embryo fibroblast (DF-1) cells, and tissue distribution and immunogenicity in specific pathogen-free (SPF) day-old chickens. APMV-3 strain Netherlands showed highest growth rate and GFP expression level among the three APMV vectors in vitro. APMV-3 strain Wisconsin and APMV-1 strain LaSota vectors were mainly confined to the trachea after vaccination of day-old SPF chickens without any observable pathogenicity, whereas APMV-3 strain Netherlands showed wide tissue distribution in different body organs (brain, lungs, trachea, and spleen) with mild observable pathogenicity. In terms of immunogenicity, both APMV-3 strain-vaccinated groups showed HI titers two to three fold higher than that induced by APMV-1 strain LaSota vaccinated group. This study offers a novel paramyxovirus vector (APMV-3 strain Wisconsin) which can be used safely for vaccination of young chickens as an alternative for APMV-1 strain LaSota vector.
创建了一个用于禽副黏病毒 3 型（APMV-3）威斯康星株的反向遗传学系统，并从基于质粒的病毒抗原基因组 cDNA 中回收了感染性病毒。将绿色荧光蛋白（GFP）基因克隆到重组 APMV-3 基因组中作为外源基因。回收病毒的 GFP 稳定表达至少连续传代 10 代。将 APMV-3 威斯康星株与 APMV-3 荷兰株和 APMV-1 拉索塔株进行了比较，作为疫苗载体。比较了三种表达 GFP 作为外源蛋白的病毒载体的 GFP 表达水平、在鸡胚成纤维细胞（DF-1）中的生长速度以及在无特定病原体（SPF）日龄鸡中的组织分布和免疫原性。在体外，三种 APMV 载体中，APMV-3 荷兰株的生长速度和 GFP 表达水平最高。接种 SPF 日龄鸡后，APMV-3 威斯康星株和 APMV-1 拉索塔株载体主要局限于气管，没有明显的致病性，而 APMV-3 荷兰株载体在不同的身体器官（脑、肺、气管和脾脏）中分布广泛，具有轻微的可见致病性。在免疫原性方面，APMV-3 株接种组的 HI 滴度均比 APMV-1 株拉索塔接种组高 2-3 倍。本研究提供了一种新型副黏病毒载体（APMV-3 威斯康星株），可安全用于小鸡接种，作为 APMV-1 拉索塔载体的替代物。
