PRRS Research Laboratory, Viral Diseases Division, Animal and Plant Quarantine Agency, Gimcheon, 39660, Republic of Korea.
Division of Animal Care, Yonam College, 313, Yeonam-ro, Seonghwan-eup, Seobuk-gu, Cheonan-si, Chungcheongnam-do, Republic of Korea.
J Gen Virol. 2023 Aug;104(8). doi: 10.1099/jgv.0.001889.
Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important virus within the swine industry. The virus causes respiratory disease and reproductive failure. Two species of PRRSV-I and II are co-dominant, yet no effective vaccination strategy has been developed to protect against these two types. With an aim to develop a chimeric vaccine strain to protect against both types, in this study, a chimeric porcine reproductive and respiratory syndrome virus (PRRSV) type I and II was rescued using reverse genetics for the first time. Four chimeric infectious clones were designed based on the genomic arrangement of the structural proteins. However, only the clone carrying the transcriptional regulatory sequence (TRS) and ORF6 of a PRRSV-I and ORF6 of a PRRSV-II generated a viable recombinant virus, suggesting that concurrent expression of ORF6 from both parental viruses is essential for the recovery of type I and II chimeric PRRSV. The chimeric virus showed significantly lower replication ability than its parental strains , which was improved by serial passaging. , groups of pigs were inoculated with either the chimeric virus, one of the parental strains, or PBS. The chimeric virus replicated in pig tissue and was detected in serum 7 days post-inoculation. Serum neutralization tests indicated that pigs inoculated with the chimeric virus elicited neutralizing antibodies that inhibited infection with strains of both species and with greater coverage than the parental viruses. In conclusion, the application of this technique to construct a chimeric PRRSV holds promise for the development of a highly effective modified live vaccine candidate. This is particularly significant since there are currently no approved commercial divalent vaccines available to combat PRRSV-I and II co-infections.
猪繁殖与呼吸综合征病毒(PRRSV)是养猪业中一种具有重要经济意义的病毒。该病毒可引起呼吸道疾病和繁殖障碍。PRRSV-I 和 II 两种病毒同时存在,但目前尚无有效的疫苗接种策略来预防这两种病毒。本研究旨在开发一种嵌合疫苗株以预防这两种类型的病毒,首次使用反向遗传学技术拯救了一种 PRRSV-I 和 II 型嵌合病毒。根据结构蛋白的基因组排列设计了四个嵌合感染性克隆,但只有携带 PRRSV-I 的转录调控序列(TRS)和 ORF6 以及 PRRSV-II 的 ORF6 的克隆能够产生有活力的重组病毒,这表明同时表达两种亲本病毒的 ORF6 对于 I 型和 II 型嵌合 PRRSV 的恢复是必需的。嵌合病毒的复制能力明显低于其亲本株,通过连续传代可以提高。将猪分为几组,分别接种嵌合病毒、亲本株之一或 PBS。嵌合病毒在猪组织中复制,并在接种后 7 天从血清中检测到。血清中和试验表明,接种嵌合病毒的猪产生了中和抗体,可抑制两种病毒株的感染,并且比亲本株的覆盖面更广。总之,该技术用于构建嵌合 PRRSV 具有开发高效改良活疫苗候选物的应用前景。由于目前尚无批准的用于对抗 PRRSV-I 和 II 混合感染的商业二价疫苗,这一点尤其重要。