Abstract：CD163 is a macrophage-specific protein in the cysteine superfamily rich in scavenger receptors and is the most important cellular receptor of Porcine reproductive and respiratory syndrome virus (PRRSV). Induced pluripotent stem cells (iPSCs) are pluripotent stem cells with biological characteristics of embryonic stem cells obtained by reprogramming after introducing foreign transcription factors into somatic cells. Firstly, porcine (Sus scrofa) CD163 gene was transferred into porcine induced pluripotent stem cells (piPSCs) by lentiviral vector system, and an induced pluripotent stem cell line stably expressing CD163 was established, which was named CD163OE-piPSC. The susceptibility of the cell line to PRRSV was verified by virus infection, and the pluripotency and proliferation of iPSCs were not affected by alkaline phosphatase staining, cell population doubling time test and 5-ethynyl-2'- deoxyuridine (EdU) cell proliferation test.The cell lines obtained in this study could be effectively used to study the pathogenesis of PRRSV-host interaction and screen the target molecules of viral diseases.
[1] 段安琴, 王宁, 于童等. 2014. 四环素调控基因(TetO)诱导重编程的猪体细胞系建立[J]. 农业生物技术学报, 22(09): 1065-1073. (Duan A Q, Wang N, Yu T, et al.2014. A porcine (Sus scrofa domesticus) somatic cell line of tetracycline operator (Tet0)-inducible system for reprogramming[J]. Journal of Agricultural Biotechnology, 22(09): 1065-1073.) [2] An T Q, Li J N, Su C M, et al.2020. Molecular and cellular mechanisms for PRRSV pathogenesis and host response to infection[J]. Virus Research, 286: 197980. [3] Burkard C, Lillico S G, Reid E, et al.2017. Precision engineering for PRRSV resistance in pigs: Macrophages from genome edited pigs lacking CD163 SRCR5 domain are fully resistant to both PRRSV genotypes while maintaining biological function[J]. PLOS Pathogens, 13(2): e1006206. [4] Ezashi T, Yuan Y, Roberts R M.2016. Pluripotent stem cells from domesticated mammals[J]. Annual Review of Animal Biosciences, 4: 223-253. [5] Fang J, Qiao S, Wang K, et al.2021. Quantitative proteomic analysis of global protein acetylation in PRRSV-infected pulmonary alveolar macrophages[J]. Proteomics, 21(2): e2000019. [6] Li L, Wu C, Hou G, et al.2017. Generation of murine macrophage-derived cell lines expressing porcine CD163 that support Porcine reproductive and respiratory syndrome virus infection[J]. BMC Biotechnology, 17: 77. [7] Montaner-Tarbes S, Del Portillo H A, Montoya M, et al.2019. Key gaps in the knowledge of the Porcine respiratory reproductive syndrome virus (PRRSV)[J]. Frontiers in Veterinary Science, 6: 38. [8] Nielsen M C, Hvidbjerg Gantzel R, Clària J, et al.2020. Macrophage activation markers, CD163 and CD206, in acute-on-chronic liver failure[J]. Cells, 9(5): 1175. [9] Sharma A, Sances S, Workman M J, et al.2020. Multi-lineage human iPSC-derived platforms for disease modeling and drug discovery[J]. Cell Stem Cell, 26(3): 309-329. [10] Skytthe M K, Graversen J H, Moestrup S K.2020. Targeting of CD163 macrophages in inflammatory and malignant diseases[J]. International Journal of Molecular Sciences, 21(15): 5497. [11] Thanawongnuwech R, Thacker E L, Halbur P G.1997. Effect of Porcine reproductive and respiratory syndrome virus (PRRSV) (isolate ATCC VR-2385) infection on bactericidal activity of porcine pulmonary intravascular macrophages (PIMs): In vitro comparisons with pulmonary alveolar macrophages (PAMs)[J]. Veterinary Immunology and Immunopathology, 59(3-4): 323-335. [12] Wang G, Yu Y, Cai X, et al.2020. Effects of PRRSV infection on the porcine thymus[J]. Trends in Microbiology, 28(3): 212-223. [13] Wang X, Wei R, Li Q, et al.2013. PK-15 cells transfected with porcine CD163 by PiggyBac transposon system are susceptible to Porcine reproductive and respiratory syndrome virus[J]. Journal of Virological Methods, 193(2): 383-390. [14] Wu X L, Zhu Z S, Xiao X, et al.2021. Inhibits family phosphatases and activates MAPK signaling pathway to maintain pluripotency in porcine induced pluripotent stem cells[J]. Zoological Research, 42(3): 377-388. [15] Xu H, Liu Z, Zheng S, et al.2020a. CD163 antibodies inhibit PRRSV infection via receptor blocking and transcription suppression[J]. Vaccines, 8(4): 592. [16] Xu K, Zhou Y, Mu Y, et al.2020b. CD163 and pAPN double-knockout pigs are resistant to PRRSV and TGEV and exhibit decreased susceptibility to PDCoV while maintaining normal production performance[J]. eLife, 9: e57132. [17] Yin B, Qi S, Sha W, et al.2021. Molecular characterization of the Nsp2 and ORF5 (ORF5a) genes of PRRSV strains in nine provinces of China during 2016-2018[J]. Frontiers in Veterinary Science, 8: 605832. [18] Yu P, Wei R, Dong W, et al.2019. CD163 MARC-145 cells resist PRRSV-2 infection via inhibiting virus uncoating, which requires the interaction of CD163 with calpain 1[J]. Frontiers in Microbiology, 10: 3115. [19] Yu S, Zhang R, Shen Q, et al.2021. ESRRB facilitates the conversion of trophoblast-like stem cells from induced pluripotent stem cells by directly regulating CDX2[J]. Frontiers in Cell and Developmental Biology, 9: 712224. [20] Zhang Y, Zhang K, Zheng H, et al.2020. Development of a monoclonal antibody against porcine CD163 SRCR5 domain which partially blocks infection of PRRSV[J]. Frontiers in Veterinary Science, 7: 597843. [21] Zhu Z, Wu X, Li Q, et al.2021. Histone demethylase complexes KDM3A and KDM3B cooperate with OCT4/SOX2 to define a pluripotency gene regulatory network[J]. FASEB Journal, 35(6): e21664.
