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Published on: May 19, 2023
Establishing Porcine Monocyte-Derived Macrophage and Dendritic Cell Systems for Studying the Interaction with PRRSV-1
Helen Singleton1, Simon P Graham1, Katherine B Bodman-Smith2
1Virology Department, Animal and Plant Health AgencySurrey, UK; Faculty of Health and Medical Sciences, University of SurreySurrey, UK.
Abstract:
Monocyte-derived macrophages (MoMØ) and monocyte-derived dendritic cells (MoDC) are two model systems well established in human and rodent systems that can be used to study the interaction of pathogens with host cells. Porcine reproductive and respiratory syndrome virus (PRRSV) is known to infect myeloid cells, such as macrophages (MØ) and dendritic cells (DC). Therefore, this study aimed to establish systems for the differentiation and characterization of MoMØ and MoDC for subsequent infection with PRRSV-1. M-CSF differentiated MoMØ were stimulated with activators for classical (M1) or alternative (M2) activation. GM-CSF and IL-4 generated MoDC were activated with the well established maturation cocktail containing PAMPs and cytokines. In addition, MoMØ and MoDC were treated with dexamethasone and IL-10, which are known immuno-suppressive reagents. Cells were characterized by morphology, phenotype, and function and porcine MØ subsets highlighted some divergence from described human counterparts, while MoDC, appeared more similar to mouse and human DCs. The infection with PRRSV-1 strain Lena demonstrated different replication kinetics between MoMØ and MoDC and within subsets of each cell type. While MoMØ susceptibility was significantly increased by dexamethasone and IL-10 with an accompanying increase in CD163/CD169 expression, MoDC supported only a minimal replication of PRRSV These findings underline the high variability in the susceptibility of porcine myeloid cells toward PRRSV-1 infection.
Insights
Porcine reproductive and respiratory syndrome virus (PRRSV) infects myeloid cells. This study established porcine monocyte-derived macrophages (MoMØ) and dendritic cells (MoDC) models, revealing varied PRRSV-1 susceptibility and replication kinetics within these cell types.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Monocyte-derived macrophages (MoMØ) and dendritic cells (MoDC) are crucial for studying host-pathogen interactions.
- Porcine reproductive and respiratory syndrome virus (PRRSV) targets myeloid cells, including macrophages and dendritic cells.
- Establishing reliable porcine myeloid cell models is essential for understanding PRRSV pathogenesis.
Purpose of the Study:
- To establish and characterize porcine MoMØ and MoDC for PRRSV-1 infection studies.
- To investigate the susceptibility and replication kinetics of PRRSV-1 in differentiated porcine myeloid cells.
- To evaluate the impact of immune-modulatory reagents on PRRSV-1 infection in these models.
Main Methods:
- Differentiation of porcine monocytes into MoMØ using M-CSF and MoDC using GM-CSF and IL-4.
- Phenotypic and functional characterization of differentiated cells.
- Infection of MoMØ and MoDC with PRRSV-1 strain Lena, with and without dexamethasone and IL-10 treatment.
Main Results:
- Porcine MoMØ and MoDC exhibited distinct characteristics, with MoDC showing more similarity to human/mouse counterparts.
- PRRSV-1 demonstrated varied replication kinetics in MoMØ and MoDC, and within their subsets.
- Dexamethasone and IL-10 increased MoMØ susceptibility to PRRSV-1, correlating with CD163/CD169 expression, while MoDC supported minimal replication.
Conclusions:
- Porcine MoMØ and MoDC serve as valuable models for PRRSV-1 research, highlighting species-specific differences.
- Myeloid cell susceptibility to PRRSV-1 infection is highly variable and influenced by activation and immune status.
- These findings contribute to understanding PRRSV pathogenesis and host immune responses in pigs.
