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
| Autor: | Nick Vereecke, Ivan Trus, Jiexiong Xie, Dayoung Oh, Sebastiaan Theuns, Jannes Sauer, Carl A. Gagnon, Nathalie Vanderheijden, Hans Nauwynck, Christian Lalonde, P Vyt, Caroline Bonckaert, Chantale Provost |
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| Přispěvatelé: | Université de Montréal. Faculté de médecine vétérinaire |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
INVOLVEMENT viruses 0403 veterinary science Drug Discovery INFECTION PK15Sn-CD163 Pharmacology (medical) Receptor NUCLEOCAPSID PROTEIN virus isolation biology 04 agricultural and veterinary sciences SYNDROME PRRS VIRUS macrophages Infectious Diseases Medicine Cellular Tropism Cell type PK15(S10-CD163) 040301 veterinary sciences PK15(Sn-CD163) Immunology PK15S10-CD163 DENDRITIC CELLS Article Virus 03 medical and health sciences NORTH-AMERICAN Sialoadhesin SIALOADHESIN Veterinary Sciences Tropism Pharmacology MARC-145 IDENTIFICATION Macrophages Porcine reproductive and respiratory syndrome virus biology.organism_classification EFFICACY Virology 030104 developmental biology Cell culture PRRSV REPLICATION Virus isolation/production production |
| Zdroj: | VACCINES Vaccines, Vol 9, Iss 594, p 594 (2021) Vaccines Volume 9 Issue 6 |
| ISSN: | 2076-393X |
| Popis: | 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., PK15Sn-CD163, PK15S10-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 (PK15Sn-CD163, PK15S10-CD163, MARC-145, and MARC-145Sn) 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 PK15Sn-CD163, PK15S10-CD163, MARC-145Sn, and MARC-145. The titers in PK15Sn-CD163 and PK15S10-CD163 cells were significantly correlated with virus titers in PAM for both PRRSV1 (p < 0.001) and PRRSV2 (p < 0.001) compared with MARC-145Sn (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, PK15Sn-CD163 and PK15S10-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-145Sn cells. The residues of PRRSV proteins that are potentially related with cell tropism will be further investigated in the future. |
| Databáze: | OpenAIRE |
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