Characterization of Monoclonal Antibodies to Measure Cell Surface Protein Levels of Human Interferon-Lambda Receptor 1.
Nicole A de WeerdOlamide OgungbolaXinyun LiuAntony Y MatthewsAmina IsmailJulian P VivianSan S LimD Lorne TyrrellNiru PutchaMike SkawinskiHarold DickensheetsThomas B LavoieRaymond P DonnellyPaul J HertzogDeanna M SanterPublished in: Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research (2023)
Type III interferons (IFN-lambdas, IFN-λs) are important antiviral cytokines that can also modulate immune responses by acting through a heterodimeric receptor composed of the specific and limited expressed IFN-λR1 chain and the ubiquitous IL-10R2 chain, which is shared with IL-10 family cytokines. Conflicting data have been reported regarding which cells express the IFN-λR1 subunit and directly respond to IFN-λs. This is, in part, owing to transcript levels of the IFN-λR1 gene, IFNLR1 , not always correlating with cell surface protein levels. In this study, we tested a panel of novel monoclonal antibodies (mAbs) that specifically recognize human IFN-λR1. Initially, antigen specificity was confirmed by enzyme-linked immunosorbent assay (ELISA), from which a subset of antibodies was selected for additional flow cytometry and neutralization assays. We further characterized two antibodies based on their strong ELISA binding activity (HLR1 and HLR14) and found only HLR14 could reliably detect cell surface IFN-λR1 protein on a variety of cell lines by flow cytometry. HLR14 could also detect IFN-λR1 protein on certain primary human blood cells, including plasmacytoid dendritic cells and B cells from peripheral blood. Availability of the HLR14 mAb will enable the quantification of IFN-λR1 protein levels on cells and better characterization of the cell specificity of the IFN-λ response.
Keyphrases
- dendritic cells
- immune response
- cell surface
- flow cytometry
- regulatory t cells
- induced apoptosis
- endothelial cells
- binding protein
- peripheral blood
- protein protein
- bone marrow
- high throughput
- stem cells
- type iii
- cell death
- monoclonal antibody
- gene expression
- signaling pathway
- cell proliferation
- oxidative stress
- single cell
- deep learning
- induced pluripotent stem cells
- inflammatory response
- rna seq
- structural basis