Generation of a porcine alveolar macrophage cell line for the growth of porcine reproductive and respiratory syndrome virus
Generation of a porcine alveolar macrophage cell line for the growth of porcine reproductive and respiratory syndrome virus
复制标题
DOI：
10.1016/j.jviromet.2009.11.003
复制
发表时间：
2010-02-01
影响因子：
3.1
通讯作者：
Lee, Changhee
中科院分区：
文献类型：
--
作者：
Lee, Yoo Jin;Park, Choi-Kyu;Lee, Changhee
猪繁殖与呼吸综合征病毒（PRRSV）对单核细胞-巨噬细胞谱系的细胞具有显著的嗜性，因此，在天然宿主中在完全分化的肺泡巨噬细胞中复制。尽管在猪肺泡巨噬细胞（PAM）上鉴定了几种假定的PRRSV受体，但发现仅CD 163能够使非允许细胞对PRRSV敏感，表明在生产性感染中需要CD 163。有趣的是，初步实验显示，以前显示不能支持PRRSV复制的永生化PAM细胞系不表达可检测水平的CD 163。这些数据表明，在连续PAM细胞系中，CD 163不可检测的表达水平与PRRSV非敏感性之间可能存在相关性。因此，在本研究中，试图用CD 163 cDNA稳定转染非允许性PAM细胞，以产生组成型表达CD 163的细胞系，并评估其对PRRSV的耐受性。新建立的PAM细胞系被证明表达稳健水平的CD 163，并且对I型和2型PRRSV毒株都是完全允许的。这种PRRSV-允许PAM细胞系将是一个有价值的工具，不仅有利于病毒的繁殖，但也提前在体外研究病毒的发病机制。(C)2009爱思唯尔有限公司版权所有。
Porcine reproductive and respiratory syndrome virus (PRRSV) has a marked tropism for cells of the monocyte-macrophage lineage and accordingly, replicates in fully differentiated alveolar macrophages in the natural host. Despite the identification of several putative receptors for PRRSV on porcine alveolar macrophages (PAM), only CD163 was found to be able to make non-permissive cells susceptible to PRRSV, indicating a requirement for CD163 in productive infection. Interestingly, the preliminary experiments revealed that the immortalized PAM cell line, which was previously shown to fail to Support PRRSV replication, does not express detectable levels of CD163. These data suggest that there may be a correlation between the CD163 undetectable expression level and PRRSV non-susceptibility in the continuous PAM cell line. In this study, therefore, it was attempted to stably transfect non-permissive PAM cells with CD163 cDNA to generate cell lines constitutively expressing CD163 and to evaluate their permissivity to PRRSV. The newly established PAM cell lines were demonstrated to express robust levels of CD163 and to be fully permissive for both type I and 2 PRRSV strains. This PRRSV-permissive PAM cell line will be a valuable tool not only to facilitate virus propagation but also to advance in vitro studies on virus pathogenesis. (C) 2009 Elsevier B.V. All rights reserved.