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A porcine alveolar macrophage cell line stably expressing CD163 demonstrates virus replication and cytokine secretion characteristics similar to primary alveolar macrophages following PRRSV infection

文献类型： 外文期刊

作者： Xu, Yu-lin 1 ; Wu, Shao-peng 1 ; Li, Yun-gang 4 ; Sun, Feng-xia 5 ; Wang, Qiu-ju 1 ; Zhao, Qing 1 ; Yu, Jiang 6 ; Tian, F 1 ;

作者机构： 1.Shandong Agr Univ, Coll Vet Med, Tai An 271000, Shandong, Peoples R China

2.Shandong Prov Key Lab Anim Biotechnol & Dis Contr, Tai An 271000, Shandong, Peoples R China

3.Minist Agr & Rural Affairs China, East China Res Ctr Anim Epidem Pathogen Biol, Tai An 271000, Shandong, Peoples R China

4.Shandong Ctr Anim Dis Control & Prevent, Jinan 250022, Peoples R China

5.Shandong Agr Univ, Coll Resources & Environm, Tai An 271000, Shandong, Peoples R China

6.Shandong Acad Agr Sci, Inst Anim Sci & Vet Med, Jinan 251000, Peoples R China

7.Shandong Key Lab Anim Dis Control & Breeding, Jinan 250100, Peoples R China

关键词： Porcine alveolar macrophage cell line; CD163; Stable expression; Cytokine; Secretion characteristics; Porcine reproductive and respiratory syndrome virus

期刊名称：VETERINARY MICROBIOLOGY （ 影响因子：3.293； 五年影响因子：3.599 ）

ISSN： 0378-1135

年卷期： 2020 年 244 卷

页码：

收录情况： SCI

摘要： The in vitro investigation of cytokine secretion induced by porcine reproductive and respiratory syndrome virus (PRRSV) requires porcine alveolar macrophages (PAMs) and their interaction with immunocytes. However, immortalized monoclonal PAMs (mPAMs) are non-permissive for PRRSV infection. The porcine CD163 receptor isolated from primary PAMs (pPAMs) confers susceptibility to PRRSV infection; thus, this approach could be used to establish a novel cell line to facilitate the exploration of PRRSV infection kinetics. Here, we amplified the coding region of the CD163 gene from pPAMs and integrated it into an mPAM line using a lentivirus expression system. After verification, the monoclonal PAM cell line stably expressing CD163 (mPAM-CD163-GFP) was infected with either the highly pathogenic PRRSV strain JXA1 or the classical PRRSV strain SD1, which produced high infectious titers of progeny virus reaching > 10(9) copies/mL or a 50 % tissue culture infective dose of 10(5.5) over at least 100 cell generations. We also investigated cytokine and Toll-like receptor expression in infected mPAM-CD163-GFP cells and pPAMs. The mPAM-CD163-GFP cell line showed similar patterns of viral replication and cytokine secretion compared with pPAMs, so it may be extremely useful for replacing primary cells for in vitro investigations of the mechanisms of cytokine secretion and interactions between PRRSV-infected PAMs and immunocytes.

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