Abstract
Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most devastating pathogens in the swine industry worldwide. Due to the lack of robust cell lines and small animal models, the pathogenesis of PRRSV infection and mechanism for protective vaccination are still not yet well understood. To obtain useful cell lines, several groups have attempted to construct different transgenic cell lines with three PRRSV receptors: CD163, CD169, and CD151. The results showed that CD163 is essential for PRRSV entry into target cells and replication, and both CD169 and CD151 play key roles during PRRSV infection. However, their interplay and combined effect remains unclear. In this study, we generated transgenic BHK-21 derived cell lines co-expressing different combinations of the three receptors, which were transfected with CD163 alone, or the combination of CD163 and CD169, or the combination of CD163 and CD151, or the combination of CD163, CD169, and CD151 using the PiggyBac transposon system. Our results showed that the synergistic interaction among the three receptors was important to improve the susceptibility of cells during PRRSV infection. Through a series of comparable analyses, we confirmed that the cell line co-expressing triple receptors sustained viral infection and replication, and was superior to the current cell platform used for the PRRSV study, MARC-145 cells. Moreover, we found that PRRSV infection of the transgenic cell lines could trigger IFN-stimulated gene responses similar to those of porcine alveolar macrophages and MARC-145 cells. In summary, we developed a stable transgenic cell line susceptible to PRRSV, which may not only provide a useful tool for virus propagation, vaccine development, and pathogenesis studies, but also establish the foundation for small animal model development.
Highlights
Porcine reproductive and respiratory syndrome (PRRS) is one of the most serious contagious diseases in pigs and causes significant economic losses in the global swine industry [1,2]
We found that the BHK-21 cell line co-expressing all three receptors was more susceptible to Porcine reproductive and respiratory syndrome virus (PRRSV) than MARC-145 cells
BHK-21-DTG2 and BHK21-TTG clones were sorted by flow cytometry for EGFP and the purity was above 95% (Fig 1B)
Summary
Porcine reproductive and respiratory syndrome (PRRS) is one of the most serious contagious diseases in pigs and causes significant economic losses in the global swine industry [1,2]. Generate a BHK-21 Derived Cell Line Susceptible for PRRSV Infection no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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