# Modulatory Effects of the Recombinant Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike S1 Subunit Protein on the Phenotype of Camel Monocyte-Derived Macrophages

## TL;DR

This study shows that the MERS-CoV S1 protein changes camel macrophages into an anti-inflammatory type, which may help explain why camels tolerate the virus better.

## Contribution

The first in vitro generation of camel monocyte-derived macrophages and the discovery of MERS-CoV S1's effect on their polarization.

## Key findings

- MERS-CoV S1 protein induces an M2-like macrophage phenotype in camels.
- MERS-CoV S1-polarized macrophages show increased phagocytosis activity.
- LPS/GM-CSF stimulation leads to an M1 macrophage phenotype in camel MDMs.

## Abstract

The current study represents the first report on the in vitro generation and polarization of monocyte-derived macrophages (MDMs) in camels and the impact of the MERS-CoV S1 protein on camel MDM phenotype. The results show a polarizing effect of the MERS-CoV S1 protein on camel MDM, turning it into an anti-inflammatory M2-like phenotype with enhanced phagocytosis activity.

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is an emerging zoonotic pathogen with different pathogenesis in humans and camels. The mechanisms behind the higher tolerance of camels to MERS-CoV infection are still unknown. Monocytes are innate myeloid cells that are able, depending on the local stimulation in their microenvironment, to differentiate into different functional subtypes of macrophages with an impact on the adaptive immune response. Several in vitro protocols have been used to induce the differentiation of monocyte-derived macrophages (MDMs) in human and several veterinary species. Such protocols are not available for camel species. In the present study, monocytes were separated from camel blood and differentiated in vitro in the presence of different stimuli into MDM. Camel MDMs generated in the presence of a combined stimulation of monocytes with LPS and GM-CSF resulted in the development of an M1 macrophages phenotype with increased abundance of the antigen-presentation receptor MHCII molecules and a decreased expression of the scavenger receptor CD163. The expression pattern of the cell markers CD163, CD14, CD172a, CD44, and CD9 on MDM generated in the presence of the MERS-CoV S1 protein revealed similarity with M-CSF-induced MDM, suggesting the potential of the MERS-CoV S1 protein to induce an M2 macrophages phenotype. Similarly to the effect of M-CSF, MERS-CoV-S protein-induced MDMs showed enhanced phagocytosis activity compared to non-polarized or LPS/GM-CSF-polarized MDMs. Collectively, our study represents the first report on the in vitro generation of monocyte-derived macrophages (MDMs) in camels and the characterization of some phenotypic and functional properties of camel MDM under the effect of M1 and M2 polarizing stimuli. In addition, the results suggest a polarizing effect of the MERS-CoV S1 protein on camel MDMs, developing an M2-like phenotype with enhanced phagocytosis activity. To understand the clinical relevance of these in vitro findings on disease pathogenesis and camel immune response toward MERS-CoV infection, further studies are required.