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Updated: Feb 3, 2026

High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes
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High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes

Published on: March 24, 2015

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Species Specificity of Type III Interferon Activity and Development of a Sensitive Luciferase-Based Bioassay for

Sophie Jacobs1, Fanny Wavreil1, Bert Schepens2,3

  • 11 de Duve Institute , Université Catholique de Louvain, Brussels, Belgium .

Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research
|October 19, 2018
PubMed

Insights

Type III interferons (IFN-λ) show some species specificity, contrary to previous beliefs. A new reporter cell line, Fawa-λ-luc, offers sensitive detection of IFN-λ in biological samples.

Area of Science:

  • Immunology
  • Virology
  • Molecular Biology

Background:

  • The type III interferon (IFN-λ) family comprises four subtypes in humans and two in mice.
  • Type I interferons (IFN-α/β) are known for strong species specificity.
  • Previous studies suggested cross-species activity for type III interferons.

Purpose of the Study:

  • To re-evaluate the cross-species activity of mouse and human type III interferons.
  • To develop a sensitive and specific bioassay for detecting type III interferons in biological fluids.

Main Methods:

  • Comparative analysis of mouse and human IFN-λ activity in different cell types.
  • Development of a firefly luciferase reporter cell line (Fawa-λ-luc) by modifying mouse epithelial cells.
  • Inactivation of Ifnar2 gene to remove type I IFN response and overexpression of mouse IFNLR1 to enhance type III IFN response in reporter cells.
  • Validation of the Fawa-λ-luc assay against ELISA for detecting mouse IFN-λ in various biological samples.

Main Results:

  • Mouse and human type III interferons exhibit some species specificity, less pronounced than type I interferons.
  • Mouse IFN-λ3 shows significant species specificity, being 25-fold less active in human cells compared to mouse IFN-λ2.
  • Amino acid differences in α helices A and F likely contribute to mouse IFN-λ3 specificity.
  • Human IFN-λ4 retains high activity in mouse cells.
  • The Fawa-λ-luc reporter cell line demonstrates high sensitivity and specificity for detecting mouse and human IFN-λ in cell culture supernatants, serum, and bronchoalveolar lavage samples.
  • The assay's sensitivity is comparable to commercial ELISA kits.

Conclusions:

  • Type III interferons possess a degree of species specificity, influenced by specific amino acid residues.
  • The Fawa-λ-luc reporter cell line provides a sensitive and specific tool for quantifying type III interferon activity in preclinical and clinical settings.
  • This assay facilitates the study of type III interferon responses in various physiological and pathological conditions.
Keywords:
IFN-λIFN-λ receptorantiviralcross-species activitycytokinerespiratory syncytial virusspecies-specific activitytype I IFN

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