Henan Key Laboratory of Immunology and Targeted Therapy, Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang, Henan, 453003, China.
Henan Key Laboratory of Immunology and Targeted Therapy, Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang, Henan, 453003, China; Department of Clinical Laboratory, Xinxiang Central Hospital, Xinxiang, Henan, 453000, China.
J Virol Methods. 2019 Dec;274:113727. doi: 10.1016/j.jviromet.2019.113727. Epub 2019 Sep 4.
CD163 is a cellular receptor for Porcine reproductive and respiratory syndrome virus (PRRSV). Transgenic expression of CD163 can predispose a variety of PRRSV non-permissive cells to PRRSV infection. These resulting cells can then be used for PRRSV production and the study of PRRSV biology. The PiggyBac (PB) transposon is a non-viral, plasmid-based mobile genetic element that can be used for gene delivery into mammalian cells. In this study, a simple and efficient method for the transfection of the porcine CD163 transgene into an immortalized porcine alveolar macrophage cell line (3D4/21), a non-permissive cell line to PRRSV infection, by PB transposition was demonstrated. The resultant stably transformed 3D4/21/CD163 cells expressed CD163 constitutively and were shown to be fully permissive for PRRSV-2 strains and yielded an excess of 10 TCID/mL of progeny virus. The PRRSV replicated more efficiently in the 3D4/21/CD163 cells than in Marc-145 cells, and the titers of the progeny PRRSV produced in the 3D4/21/CD163 cells were higher than those produced in Marc-145 cells. This simplified PB transposon-generated PRRSV-2 permissive 3D4/21/CD163 cell line could facilitate PRRSV production and accelerate the study of virus-host interactions in vitro.
CD163 是猪繁殖与呼吸综合征病毒（PRRSV）的细胞受体。CD163 的转基因表达可使多种 PRRSV 非允许细胞易于感染 PRRSV。这些产生的细胞随后可用于 PRRSV 的生产和 PRRSV 生物学的研究。PiggyBac（PB）转座子是一种非病毒、基于质粒的可移动遗传元件，可用于将基因递送至哺乳动物细胞。在这项研究中，展示了一种简单有效的方法，通过 PB 转座将猪 CD163 转基因转染到一种永生化的猪肺泡巨噬细胞系（3D4/21）中，该细胞系对 PRRSV 感染不允许。所得的稳定转化的 3D4/21/CD163 细胞持续表达 CD163，并且对 PRRSV-2 株完全允许，产生超过 10 TCID/mL 的子代病毒。PRRSV 在 3D4/21/CD163 细胞中的复制效率高于 Marc-145 细胞，并且在 3D4/21/CD163 细胞中产生的子代 PRRSV 的滴度高于在 Marc-145 细胞中产生的滴度。这种简化的 PB 转座子生成的 PRRSV-2 允许的 3D4/21/CD163 细胞系可以促进 PRRSV 的生产，并加速病毒-宿主相互作用的体外研究。
