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

Author:

Wu Xiangju12ORCID,Qi Jing2,Cong Xiaoyan2,Chen Lei2,Hu Yue2,Yoo Dongwan3ORCID,Wang Guisheng4,Tian Fulin4,Li Feng5,Sun Wenbo2,Chen Zhi2,Guo Lihui2,Wu Jiaqiang2,Li Jun2,Wang Jinbao12ORCID,Zhao Xiaomin1ORCID,Du Yijun2ORCID

Affiliation:

1. Key Laboratory of Animal Biotechnology and Disease Control and Prevention of Shandong Province, College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai’an 271018, China

2. Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Sangyuan Road No. 8, Jinan 250100, China

3. Department of Pathobiology, University of Illinois at Urbana-Champaign, 2001 South Lincoln Ave, Urbana, IL 61802, USA

4. Shandong Provincial Center for Animal Disease Control and Prevention, Jinan 250022, China

5. Department of Biology and Microbiology and Department of Veterinary and Biomedical Sciences, South Dakota State University, Brookings, SD 57007, USA

Abstract

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.

Funder

National Key Research and Development Program of China

Publisher

Hindawi Limited

Subject

General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Medicine

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