Division of Biological Products, Indian Council of Agricultural Research (ICAR)-Indian Veterinary Research Institute (IVRI), Izatnagar, 243122, Bareilly, Uttar Pradesh, India; ICAR -National Research Centre on Pig, Rani, Guwahati, Assam, 781131, India.
Division of Biological Products, Indian Council of Agricultural Research (ICAR)-Indian Veterinary Research Institute (IVRI), Izatnagar, 243122, Bareilly, Uttar Pradesh, India.
Microb Pathog. 2021 Jul;156:104940. doi: 10.1016/j.micpath.2021.104940. Epub 2021 May 5.
Cellular receptors play an important role in entry and cell to cell spread of morbillivirus infections. The cells expressing SLAM and Nectin-4 have been used for successful and efficient isolation of canine distemper virus (CDV) in high titre. There are several methods for generation of cells expressing receptor molecules. Here, we have used a comparatively cheaper and easily available method, pcDNA 3.1 (+) for engineering Vero cells to express SLAM gene of goat, sheep and dog origin (Vero/Goat/SLAM (VGS), Vero/Sheep/SLAM (VSS) and Vero/Dog/SLAM (VDS), respectively). The generated cell lines were then compared to test their efficacy to support CDV replication. CDV could be grown in high titre in the cells expressing SLAM and a difference of log two could be recorded in virus titre between VDS and native Vero cells. Also, CDV could be grown in a higher titre in VDS as compared to VGS and VSS. The finding of this study supports the preferential use of SLAM expressing cells over the native Vero cells by CDV. Further, the higher titre of CDV in cells expressing dog-SLAM as compared to the cells expressing SLAM of non-CDV hosts (i.e. goat and sheep) points towards the preferential use of dog SLAM by the CDV and may be a plausible reason for differential susceptibility of small ruminants and Canines to CDV.
细胞受体在麻疹病毒感染的进入和细胞间传播中起着重要作用。表达 SLAM 和 Nectin-4 的细胞已被用于成功和有效地分离高滴度的犬瘟热病毒 (CDV)。有几种方法可以生成表达受体分子的细胞。在这里,我们使用了一种相对便宜且易于获得的方法 pcDNA 3.1(+),用于工程化表达山羊、绵羊和犬源性 SLAM 基因的 Vero 细胞 (Vero/Goat/SLAM(VGS)、Vero/Sheep/SLAM(VSS)和 Vero/Dog/SLAM(VDS))。然后比较生成的细胞系,以测试它们支持 CDV 复制的效果。CDV 可以在表达 SLAM 的细胞中以高滴度生长,并且在 VDS 和天然 Vero 细胞之间可以记录到病毒滴度的对数差异为 2。此外,与 VGS 和 VSS 相比,CDV 可以在 VDS 中以更高的滴度生长。这项研究的结果支持 CDV 优先使用表达 SLAM 的细胞而不是天然的 Vero 细胞。此外,与非 CDV 宿主(即山羊和绵羊)表达 SLAM 的细胞相比,犬 SLAM 表达细胞中 CDV 的滴度更高,这表明 CDV 优先使用犬 SLAM,这可能是小反刍动物和犬类对 CDV 易感性不同的一个合理原因。