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Characterization of Monoclonal Antibodies to Measure Cell Surface Protein Levels of Human Interferon-Lambda Receptor 1
de Weerd, Nicole A. Ogungbola, Olamide Liu, Xinyun Matthews, Antony Y. Ismail, Amina Vivian, Julian Philip Lim, San S. Tyrrell, D. Lorne Putcha, Niru Skawinski, Mike
de Weerd, Nicole A.
Ogungbola, Olamide
Liu, Xinyun
Matthews, Antony Y.
Ismail, Amina
Vivian, Julian Philip
Lim, San S.
Tyrrell, D. Lorne
Putcha, Niru
Skawinski, Mike
Abstract
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.
Keywords
type III interferons, flow cytometry, antiviral, IFN receptors
Date
2023
Type
Journal article
Journal
Book
Volume
43
Issue
9
Page Range
403-413
Article Number
ACU Department
Marketing and Communications
Research Office
Research Office
Collections
Relation URI
Event URL
Open Access Status
License
All rights reserved
File Access
Controlled
Notes
Copyright 2023, Mary Ann Liebert, Inc., publishers.