Comparison of Primary Virus Isolation in Pulmonary Alveolar Macrophages and Four Different Continuous Cell Lines for Type 1 and Type 2 Porcine Reproductive and Respiratory Syndrome Virus

Jiexiong Xie; Nick Vereecke; Sebastiaan Theuns; Dayoung Oh; Nathalie Vanderheijden; Ivan Trus; Jannes Sauer; Philip Vyt; Caroline Bonckaert; Christian Lalonde; Chantale Provost; Carl A Gagnon; Hans Nauwynck

doi:10.3390/vaccines9060594

Comparison of Primary Virus Isolation in Pulmonary Alveolar Macrophages and Four Different Continuous Cell Lines for Type 1 and Type 2 Porcine Reproductive and Respiratory Syndrome Virus

Vaccines (Basel). 2021 Jun 3;9(6):594. doi: 10.3390/vaccines9060594.

Authors

Jiexiong Xie¹, Nick Vereecke^{1

2}, Sebastiaan Theuns², Dayoung Oh¹, Nathalie Vanderheijden¹, Ivan Trus³, Jannes Sauer², Philip Vyt⁴, Caroline Bonckaert⁵, Christian Lalonde⁶, Chantale Provost⁶, Carl A Gagnon⁶, Hans Nauwynck^{1

2}

Affiliations

¹ Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium.
² PathoSense BV, 2500 Lier, Belgium.
³ Vaccine and Infectious Disease Organization (VIDO)-International Vaccine Centre, University of Saskatchewan, Saskatoon, SK S7N 5E3, Canada.
⁴ Dialab, Belsele, 9111 Sint-Niklaas, Belgium.
⁵ Animal Health Care Flanders (DGZ), 8820 Torhout, Belgium.
⁶ Swine and Poultry Infectious Diseases Research Center (CRIPA)-Fonds de Recherche du Québec, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, QC J2S 2M2, Canada.

Abstract

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) has a highly restricted cellular tropism. In vivo, the virus primarily infects tissue-specific macrophages in the nose, lungs, tonsils, and pharyngeal lymphoid tissues. In vitro however, the MARC-145 cell line is one of the few PRRSV susceptible cell lines that are routinely used for in vitro propagation. Previously, several PRRSV non-permissive cell lines were shown to become susceptible to PRRSV infection upon expression of recombinant entry receptors (e.g., PK15^Sn-CD163, PK15^S10-CD163). In the present study, we examined the suitability of different cell lines as a possible replacement of primary pulmonary alveolar macrophages (PAM) cells for isolation and growth of PRRSV. The susceptibility of four different cell lines (PK15^Sn-CD163, PK15^S10-CD163, MARC-145, and MARC-145^Sn) for the primary isolation of PRRSV from PCR positive sera (both PRRSV1 and PRRSV2) was compared with that of PAM. To find possible correlations between the cell tropism and the viral genotype, 54 field samples were sequenced, and amino acid residues potentially associated with the cell tropism were identified. Regarding the virus titers obtained with the five different cell types, PAM gave the highest mean virus titers followed by PK15^Sn-CD163, PK15^S10-CD163, MARC-145^Sn, and MARC-145. The titers in PK15^Sn-CD163 and PK15^S10-CD163 cells were significantly correlated with virus titers in PAM for both PRRSV1 (p < 0.001) and PRRSV2 (p < 0.001) compared with MARC-145^Sn (PRRSV1: p = 0.22 and PRRSV2: p = 0.03) and MARC-145 (PRRSV1: p = 0.04 and PRRSV2: p = 0.12). Further, a possible correlation between cell tropism and viral genotype was assessed using PRRSV whole genome sequences in a Genome-Wide-Association Study (GWAS). The structural protein residues GP2:187L and N:28R within PRRSV2 sequences were associated with their growth in MARC-145. The GP5:78I residue for PRRSV2 and the Nsp11:155F residue for PRRSV1 was linked to a higher replication on PAM. In conclusion, PK15^Sn-CD163 and PK15^S10-CD163 cells are phenotypically closely related to the in vivo target macrophages and are more suitable for virus isolation and titration than MARC-145/MARC-145^Sn cells. The residues of PRRSV proteins that are potentially related with cell tropism will be further investigated in the future.

Keywords: MARC-145; PK15S10-CD163; PK15Sn-CD163; PRRSV; macrophages; virus isolation/production.