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Updated: Feb 14, 2026

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A cell-based interferon-tau assay with an interferon-stimulated gene 15 promoter

Noriyuki Toji1,2, Katsuo Koshi1, Tadashi Furusawa3

  • 1Cooperative Department of Veterinary Medicine, Faculty of Agriculture, Iwate University.

Biomedical Research (Tokyo, Japan)
|February 23, 2018
PubMed

Insights

Researchers developed a new cell-based assay for detecting interferon-tau (IFNT), a key pregnancy signal in cows. This convenient system aids in diagnosing early pregnancy and assessing bovine reproductive health.

Area of Science:

  • Veterinary Science
  • Reproductive Biology
  • Biotechnology

Background:

  • Interferon-tau (IFNT) is crucial for early pregnancy recognition in ruminants.
  • Accurate detection of IFNT is needed for diagnosing bovine endometrial and trophoblastic functions, including pregnancy status.

Purpose of the Study:

  • To develop a novel, convenient cell-based assay for quantifying interferon-tau (IFNT) activity in cows.
  • To establish a reporter cell line for sensitive detection of IFNT and other type I interferons.

Main Methods:

  • Developed a luciferase reporter system by stably integrating an interferon-stimulated gene promoter into Madin-Darby bovine kidney (MDBK) cells.
  • Utilized reverse transcription-quantitative PCR to confirm the reactivity of interferon-stimulated genes to IFNT.
  • Identified the ISG15 gene promoter for optimal responsiveness to IFNT and other IFNs using luciferase assays.
  • Employed the PiggyBac vector system for stable transfection of the reporter gene into MDBK cells.

Main Results:

  • Confirmed reactivity of four interferon-stimulated genes to IFNT in MDBK cells.
  • Identified the ISG15 upstream region as a highly responsive promoter for IFNT detection.
  • Established a stable MDBK cell line that exhibits a dose-dependent response to type I IFNs.
  • Demonstrated the cell line's suitability for quantifying IFNT and other type I IFNs.

Conclusions:

  • A novel, convenient cell-based assay for IFNT detection has been successfully developed.
  • The established reporter cell line provides a sensitive and dose-dependent method for quantifying type I IFNs.
  • This system holds significant potential for improving the diagnosis of reproductive functions and gestation status in cows.

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