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A novel method for detection of IFN-lambda 3 binding to cells for quantifying IFN-lambda receptor expression.

Deanna M Santer1, Gillian E S Minty1, Adil Mohamed1

  • 1Li Ka Shing Institute of Virology and Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, Canada.

Journal of Immunological Methods
|March 10, 2017
PubMed
Summary
This summary is machine-generated.

Type III interferons (IFN-lambdas) are crucial antiviral cytokines. This study developed a new assay to measure IFN-lambda 3 binding to its receptor, revealing cell-specific responses and quantifying receptor expression.

Keywords:
Binding assayFlow cytometryInterferon-lambdaInterferon-lambda receptorInterferon-stimulated genes

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Area of Science:

  • Immunology
  • Virology
  • Cell Biology

Background:

  • Type III interferons (IFN-lambdas) are critical antiviral cytokines that modulate immune responses via the IFN-λR1/IL-10R2 receptor.
  • Previous studies have yielded conflicting data regarding which cells express IFN-λR1 and respond to IFN-lambdas.

Purpose of the Study:

  • To develop and validate a novel method for measuring IFN-lambda 3 binding to the IFN-λR1/IL-10R2 receptor on various cell types.
  • To correlate IFN-lambda 3 binding affinity with functional interferon-stimulated gene (ISG) activity.

Main Methods:

  • Development of a novel binding assay to quantify IFN-lambda 3 interaction with IFN-λR1/IL-10R2 on cell surfaces.
  • Assessment of ISG induction as a functional readout of IFN-lambda 3 activity in multiple cell lines (Huh7.5, Raji, Jurkat, U937).
  • Validation using IFNLR1 knockdown in Huh7.5 cells to confirm assay specificity and functional correlation.

Main Results:

  • Huh7.5 hepatoma cells exhibited the highest IFN-lambda 3 binding affinity (lowest Kd(app)) and ISG induction.
  • Raji (B cells) and Jurkat (T cells) showed moderate binding and lower ISG responses, while U937 cells (monocytes) displayed minimal binding and no ISG induction.
  • IFNLR1 knockdown in Huh7.5 cells significantly reduced binding and ISG induction (up to 93%).
  • Maximal ISG responses were observed at 24 hours post-IFN-lambda 3 stimulation.

Conclusions:

  • The developed binding assay accurately quantifies IFN-λ receptor surface expression across diverse cell types.
  • IFN-lambda 3 responsiveness is cell-dependent, with significant variations observed between cell lines.
  • The assay provides a reliable method to predict IFN-lambda 3 functional activity based on receptor expression levels.