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

抄録

Interferon-tau (IFNT) is known as an early pregnancy recognition signal in ruminants. An accurate and convenient IFNT detection system is desirable for the diagnosis of endometrial and trophoblastic functions, including gestation status, in cows. The aim of this study was to develop a new cell-based assay, which involved the stable introduction of an interferon-stimulated gene promoter to a luciferase reporter system. The reactivity of four interferon-stimulated genes to IFNT in Madin-Darby bovine kidney (MDBK) cells was confirmed using reverse transcription-quantitative PCR. The upstream region of the interferon-stimulated gene 15 ubiquitin-like modifier (ISG15) gene as the promoter of the reporter gene, which is more responsive to IFNT and other IFNs, was determined using the luciferase assay. The reporter gene with the ISG15 upstream region was stably transfected into MDBK cells using the PiggyBac vector system; this cell line responded to type I IFNs in a dose-dependent manner. Because of its convenience, this cell line is suitable for the quantification of IFNT as well as other type I IFNs activities.