Research & Development Center, NH Foods Ltd., 3-3 Midorigahara, Tsukuba, Ibaraki 300-2646, Japan.
Division of Animal Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, 1-2 Ohwashi, Tsukuba, Ibaraki 305-8634, Japan.
J Virol Methods. 2021 Feb;288:114026. doi: 10.1016/j.jviromet.2020.114026. Epub 2020 Nov 22.
Porcine reproductive and respiratory syndrome virus (PRRSV) displays restricted tropism to porcine alveolar macrophages in nature. Meanwhile, non-porcine cell lines derived from African green monkey kidney cell lines are permissive to PRRSV, resulting in their widespread use in PRRSV research. Furthermore, genetically modified cell lines expressing receptors targeted by PRRSV have been established. We previously established porcine immortalized kidney-derived macrophages (IPKMs) that maintained typical macrophage function. In the present study, we demonstrated the advantages of IPKMs for PRRSV research. IPKMs expressed receptors for PRRSV such as CD163 and CD169. The efficiency of virus isolation from field biological samples was higher for IPKMs than for MARC-145 cells. Five different clusters of North American type PRRSV were propagated in IPKMs. Four field strains continuously produced progeny viruses during 10 continuous passages. The efficiency of virus isolation from field biological samples and continuous progeny virus production in the sequential passages using IPKMs indicated that these cells are good vessels for PRRSV research.
猪繁殖与呼吸综合征病毒(PRRSV)在自然界中对猪肺泡巨噬细胞具有有限的嗜性。同时,来源于非洲绿猴肾细胞系的非猪源细胞系也允许 PRRSV 感染,这导致它们在 PRRSV 研究中被广泛应用。此外,还建立了表达 PRRSV 靶向受体的基因修饰细胞系。我们之前建立了具有典型巨噬细胞功能的猪永生化肾源性巨噬细胞(IPKMs)。在本研究中,我们证明了 IPKMs 在 PRRSV 研究中的优势。IPKMs 表达 PRRSV 的受体,如 CD163 和 CD169。从田间生物样本中分离病毒的效率在 IPKMs 中比在 MARC-145 细胞中更高。五种不同的北美型 PRRSV 聚类在 IPKMs 中繁殖。四个田间分离株在 10 个连续传代中连续产生子代病毒。使用 IPKMs 从田间生物样本中分离病毒和连续传代中产生子代病毒的效率表明,这些细胞是 PRRSV 研究的良好载体。