Establishment and Characterization of a High and Stable Porcine CD163-Expressing MARC-145 Cell Line.

Establishment and Characterization of a High and Stable Porcine CD163-Expressing MARC-145 Cell Line.
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高稳定表达猪 CD163 MARC-145 细胞系的建立和表征

DOI:
10.1155/2018/4315861
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发表时间:
2018
影响因子:
--
通讯作者:
Du Y
Du Y
中科院分区:
生物学3区
文献类型:
--
作者:
Wu X;Qi J;Cong X;Chen L;Hu Y;Yoo D;Wang G;Tian F;Li F;Sun W;Chen Z;Guo L;Wu J;Li J;Wang J;Zhao X;Du Y

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多种猪繁殖与呼吸综合征病毒(PRRSV)的分离鉴定对PRRSV研究和疾病管理具有重要意义。然而,PRRSV对感染具有有限的细胞趋向性。MARC-145细胞通常用于北美基因型PRRSV的分离和疫苗生产。但是MARC-145细胞有一些局限性,比如病毒产量低。CD163是一种细胞受体,在多种非允许细胞系中介导PRRSV的生产性感染。在这项研究中,我们建立了一个高稳定表达CD163- (pCD163-)的猪MARC-145细胞系,以提高其对PRRSV感染的易感性。间接免疫荧光法(IFA)和Western blot检测显示pCD163在pCD163- marc细胞株中的表达高于MARC-145细胞。与MARC-145细胞相比,pCD163-MARC细胞系增殖PRRSV的能力显著提高。最后,我们发现pCD163-MARC细胞株对临床PRRSV样本的分离率更高,并且比MARC-145细胞更有效地繁殖PRRSV减毒活疫苗株。该pCD163-MARC细胞系将为PRRSV的繁殖和研究提供有价值的工具。
Isolation and identification of diverse porcine reproductive and respiratory syndrome viruses (PRRSVs) play a fundamental role in PRRSV research and disease management. However, PRRSV has a restricted cell tropism for infection. MARC-145 cells are routinely used for North American genotype PRRSV isolation and vaccine production. But MARC-145 cells have some limitations such as low virus yield. CD163 is a cellular receptor that mediates productive infection of PRRSV in various nonpermissive cell lines. In this study, we established a high and stable porcine CD163- (pCD163-) expressing MARC-145 cell line toward increasing its susceptibility to PRRSV infection. Indirect immunofluorescence assay (IFA) and Western blotting assays showed that pCD163 was expressed higher in pCD163-MARC cell line than MARC-145 cells. Furthermore, the ability of pCD163-MARC cell line to propagate PRRSV was significantly increased as compared with MARC-145 cells. Finally, we found that pCD163-MARC cell line had a higher isolation rate of clinical PRRSV samples and propagated live attenuated PRRS vaccine strains more efficiently than MARC-145 cells. This pCD163-MARC cell line will be a valuable tool for propagation and research of PRRSV.
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