Type I and type III interferons (IFNs) together regulate the early response to viral infection including SARS-CoV-2. Despite displaying structural and functional similarities, type I and type III IFNs operate via different heterodimeric receptor complexes. Type I IFNs (IFNα, IFNβ, etc) bind ubiquitously expressed IFN alpha/beta receptor (IFNAR)1 and IFNAR2. In contrast, type III IFNs (IFNλ1-4) bind a unique high affinity receptor, IFN lambda receptor (IFNLR)1 which displays restricted cellular expression and a widely expressed low affinity receptor shared with the Interleukin(IL)10 cytokine superfamily (IL10R2).
Here we report that the type I IFN receptor IFNAR2 is required for mediating the anti-viral properties of the type III IFN, IFNλ1 on lung epithelial cells. Using microscale thermophoresis, we observed high affinity binding of IFNλ1 to the extracellular domain of IFNAR2; a characteristic not displayed by IFNλ3 or IFNλ4. We used direct protein-protein interactions and mass spectrometry to show that IFNAR2 co-operates with IFNLR1 to bind IFNλ1, forming a novel ternary protein complex. Furthermore, IFNλ1, but not IFNλ3, utilise IFNAR2 on the surface of lung epithelial cells for efficient expression of interferon regulated genes (IRGs) including key anti-viral genes such as MX2, OAS2, BST2, STAT1, STAT2, IFITM1 and IFITM3. Finally, we demonstrate that IFNλ1, but not IFNλ3, requires IFNAR2 on lung epithelial cells to upregulate cell-surface levels of the viral restriction factor IFITM3 and to efficiently protect cells from viral infection.
Our data shows that IFNλ1 binds IFNAR2 to form a novel protein complex that is required for efficient IFNλ1-mediated gene induction and anti-viral activity in lung epithelial cells.