The Neonatal Fc Receptor (FcRn) is a pan-arterivirus receptor
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Jul 23, 2024 version files 784.08 KB
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README.md
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Abstract
Arteriviruses infect a variety of mammalian hosts, but the receptors used by these viruses to enter cells are poorly understood. We identified the neonatal Fc receptor (FcRn) as an important pro-viral host factor via comparative genome-wide CRISPR-knockout screens with multiple arteriviruses. Using a panel of cell lines and divergent arteriviruses, we demonstrate that FcRn is required for the entry step of arterivirus infection and serves as a molecular barrier to arterivirus cross-species infection. We also show that FcRn synergizes with another known arterivirus entry factor, CD163, to mediate arterivirus entry. Overexpression of FcRn and CD163 sensitizes non-permissive cells to infection and enables the culture of fastidious arteriviruses. Treatment of multiple cell lines with a pre-clinical anti-FcRn monoclonal antibody blocked infection and rescued cells from arterivirus-induced death. Altogether, this study identifies FcRn as a novel pan-arterivirus receptor, with implications for arterivirus emergence, cross-species infection, and host-directed pan-arterivirus countermeasure development.
https://doi.org/10.5061/dryad.h9w0vt4s6
Each tab in the .csv file corresponds to the data from a figure in the corresponding manuscript. RRA= Robust Rank Aggregation; DPI= days post infection; PFU= plaque-forming units; FFU= focus forming assay; WT= wild type; FcRn=neonatal Fc receptor; EV= empty vector; YFV= yellow fever virus; SHFV= Simian Hemorrhagic Fever Virus; PRRSV= porcine respiratory and reproductive syndrome virus; EAV= equine arteritis virus; KRCV-1= Kibale Red Colobus Virus-1; PBJV= Pebjah virus; Ct= cycle threshold. All blank cells are left intentionally blank.
Figure 1A: shows PFU production of SHFV PRRSV on wild-type MA-104 cells inoculated at MOI=0.01, 2 replicates per virus.
Figure 1E: shows RRA scores from the CRISPR-KO survival screen (RRA scores generated in MAGeCK software), with SHFV RRA vs. PRRSV RRA.
Figure 1 F: shows PFU (or FFU for YFV) production (per mL of supernatant) of multiple viruses on MA-104ΔFcRn cells vs. MA-104ΔEV cells, 3 replicates per virus/cell combination.
Figure 2D: shows percent vRNA+ cells (MA-104 WT vs. MA-104ΔFcRn vs. MA-104ΔFcRn+FcRn) 24 hours post-inoculation with SHFV, 3 replicates per cell line.
Figure 2E: Percent vRNA+ cells (MA-104 WT vs. MA-104ΔFcRn vs. MA-104ΔFcRn+FcRn) at multiple time points post-inoculation with SHFV, 3 replicates per cell line per time point.
Figure 3A: PFU production (per mL of supernatant) from cells (MA-104 WT vs. MA-104ΔFcRn) transfected with SHFV genome at multiple time points, 3 replicates per cell line per time point.
Figure 3B: PFU production (per mL of supernatant) from MA-104ΔFcRn cells 48 hrs after infection vs. transfection with SHFV genome, 3 replicates per condition.
Figure 3C: RT-qPCR data examining internalization of SHFV in cells (MA-104 WT vs. MA-104ΔFcRn vs. MA-104ΔFcRn+FcRn), normalized to WT condition, 3 replicates per condition.
Figure 3D: RT-qPCR data examining internalization of SHFV in cells (Vero vs. Vero+FcRn), normalized to WT condition, 3 replicates per condition.
Figure 3E: PFU production (per mL of supernatant) from MA-104 cells incubated with a concentration range of monoclonal antibodies (IgG4 isotype control vs. orilanolimab), 2 replicates per condition.
Figure 3F: SHFV N-gene copies per mL of supernatant produced from cells (MA-104, ACHN+hCD163, iPSC-derived macrophages) incubated with monoclonal antibodies (IgG4 isotype control vs. orilanolimab), 2 replicates per condition.
Figure 3G: Ct values from KRCV-1-specific RT-qPCR produced from iPSC-derived macrophages incubated with monoclonal antibodies (IgG4 isotype control vs. orilanolimab), 2 replicates per condition.
Figure 3H: Ct values from PBJV-specific RT-qPCR produced from iPSC-derived macrophages incubated with monoclonal antibodies (IgG4 isotype control vs. orilanolimab), 2 replicates per condition.
Figure 3J: Viability with fluorescence (arbitrary units) used as an indicator (Cell-titer-glo) for iPSC-derived macrophages incubated with monoclonal antibodies (IgG4 isotype control vs. orilanolimab) and infected with multiple arteriviruses, two replicates per condition.
Figure 4C: Measures of Viral (v)RNA quantification in images (viral (v)RNA positive cells (%), Viral (v)RNA positive puncta per infected cell, and Viral (v)RNA staining intensity per infected cell) of SHFV-infected cells (MA-104 WT vs. MA-104+gFcRn vs. MA-104+CD163 vs. MA-104+gCD163+gFcRn).
Figure 4D: PFU production (per mL of supernatant) from SHFV-infected cells (MA-104 WT vs. MA-104+gCD163+gFcRn) at multiple time points following different MOIs (0.03 vs. 3), 3 replicates per condition.
Figure 4G: PFU detection when using different MA-104 cells (MA-104 WT vs. MA-104+gFcRn vs. MA-104+CD163 vs. MA-104+gCD163+gFcRn) as substrate for plaque assay, with PFU production normalized to WT cells, 3 replicates per condition.
Figure 5B: PFU production (per mL of supernatant) from SHFV-infected cells (MA-104 WT vs. MA104∆FcRN+gCD163 vs. MA104+FcRN∆gCD163) at multiple time points, 3 replicates per condition.
Figure 5C: Measures of Viral (v)RNA quantification in images (viral (v)RNA positive cells (%), Viral (v)RNA positive puncta per infected cell, and Viral (v)RNA staining intensity per infected cell) of SHFV-infected cells (MA-104 WT vs. vs. MA104∆FcRN+gCD163 vs. MA104+FcRN∆gCD163).
Figure 6B: PFU production (per mL of supernatant) from SHFV-infected cells (Vero WT vs. Vero+gCD163 vs. Vero+gFcRn vs. Vero+gCD163+gFcRn) at multiple time points, 3 replicates per condition.
Figure 7A: PFU production (per mL of supernatant) from arterivirus (SHFV, PRRSV, EAV)-infected MA-104 cells expressing various FcRn orthologs (human, pig, mouse, horse, monkey), 2 replicates per condition.
Figure 7B: PFU production (per mL of supernatant) from arterivirus (SHFV, PRRSV, EAV)-infected MA-104 cells expressing various CD163 orthologs (human, pig, mouse, horse, monkey), 2 replicates per condition.
Figure 8A: Ct values (and N-gene copies for SHFV) from arterivirus-specific RT-qPCR measuring SHFV, PBJV, and KRCV, on cells (MA-104 WT vs. MA104+gFcRN+gCD163) at various time points, 3 replicates per condition.
Figure 8B: PFU production (per mL of supernatant) from KRCV-1-infected cells (MA-104 WT vs. MA104+gFcRN+gCD163), 3 replicates per condition.