Validation of an interferon stimulatory response element reporter gene assay for quantifying type I interferons

McCoski, Sarah R.; Xie, Markus M.; Hall, Everette; Mercadante, P.M.; Spencer, Thomas E.; Lonergan, Patrick; Ealy, Alan D.

doi:10.1016/j.domaniend.2013.12.003

Cited by 8 publications

(6 citation statements)

References 24 publications

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“…The limit of detection for IFNT in the present study was 0.4 ng/mL, which is equal to approximately 8 IU/mL. The value was comparable with that in a previous report (20). These observations indicate that, after further investigation, the assay may be suitable to replace the traditional plaque assay for IFNT.…”

Section: Response Of Isg15 Gene Promoter Activity To Recombinant Ifnssupporting

confidence: 88%

“…These complexities have limited the widespread use of antiviral assays. Although reporter gene assay systems to measure IFNs have been proposed over the past 20 years as alternative methods to antiviral assays (4,5,11,18,20,28), these are not without complications, such as the use of viral vectors and the regulations of genetic modification. In the present study, we developed a novel cell-based IFNT bioassay system with ISG15 promoter-luciferase reporter genes.…”

Section: Response Of Isg15 Gene Promoter Activity To Recombinant Ifnsmentioning

confidence: 99%

“…A suitable combination of cells and promoter assay systems may also be applicable to biological functions. The application of promoter assays for the detection of IFN has been examined and suggested to be accurate (4,5,11,18,20,28). However, an accurate and convenient detection system is still desired for the diagnosis of gestation status in cows.…”

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confidence: 99%

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

et al. 2018

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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.

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Section: Response Of Isg15 Gene Promoter Activity To Recombinant Ifnssupporting

confidence: 88%

Section: Response Of Isg15 Gene Promoter Activity To Recombinant Ifnsmentioning

confidence: 99%

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

et al. 2018

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“…Individual uterine flushes were assessed for interferon-tau (IFNT) protein content by the ISRE-Luc bioassay described previously by this laboratory [ 93 ]. In brief, Madin-Darby bovine kidney cells (MDBK; ATCC#CCL-22) that were transduced with an ISRE-Luc reporter were plated into 96-well polystyrene plates with opaque walls and optically clear bottoms (Corning Inc., Corning, NY) at a density of 5–10 × 10 5 cells/well in Dulbecco’s modified eagle medium (DMEM, 25 mM glucose; Life Technologies, Grand Island, NY) containing 10% ( v /v) fetal bovine serum (FBS), and antibiotics (50 IU Penicillin G and 50 μg/ml Streptomycin sulfate).…”

Section: Methodsmentioning

confidence: 99%

Exposure to maternal obesity alters gene expression in the preimplantation ovine conceptus

et al. 2018

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BackgroundEmbryonic and fetal exposure to maternal obesity causes several maladaptive morphological and epigenetic changes in exposed offspring. The timing of these events is unclear, but changes can be observed even after a short exposure to maternal obesity around the time of conception. The hypothesis of this work is that maternal obesity influences the ovine preimplantation conceptus early in pregnancy, and this exposure will affect gene expression in embryonic and extraembryonic tissues.ResultsObese and lean ewe groups were established by overfeeding or normal feeding, respectively. Ewes were then bred to genetically similar rams. Conceptuses were collected at day 14 of gestation. Morphological assessments were made, conceptuses were sexed by genomic PCR analysis, and samples underwent RNA-sequencing analysis. While no obvious morphological differences existed between conceptuses, differentially expressed genes (≥ 2-fold; ≥ 0.2 RPKM; ≤ 0.05 FDR) were detected based on maternal obesity exposure (n = 21). Also, differential effects of maternal obesity were noted on each conceptus sex (n = 347). A large portion of differentially expressed genes were associated with embryogenesis and placental development.ConclusionsFindings reveal that the preimplantation ovine conceptus genome responds to maternal obesity in a sex-dependent manner. The sexual dimorphism in response to the maternal environment coupled with changes in placental gene expression may explain aberrations in phenotype observed in offspring derived from obese females.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-5120-0) contains supplementary material, which is available to authorized users.

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“…Concentrations of IFN-τ in the uterine flush were determined using a luciferase-based interferon stimulatory response element reporter gene assay in MDBK cells as described previously (McCoski et al, 2014). Recombinant human IFN-α (3.87 × 10 8 IU/mg; EMD Biosciences) was used to build the standard curves and 50 µL of One-Glo Luciferase Assay Substrate (Promega Corp.) was added to each well to determine luciferase activity.…”

Section: Measurement Of Ifn-τ In Uterine Flushmentioning

confidence: 99%

Effects of progesterone concentrations and follicular wave during growth of the ovulatory follicle on conceptus and endometrial transcriptome in dairy cows

Bisinotto

Ribeiro

Greco

et al. 2022

Journal of Dairy Science

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Objectives were to evaluate the effects of follicular wave and progesterone concentration on growth of the ovulatory follicle, conceptus elongation, uterine IFN-τ concentration, and transcriptome of conceptus and endometrium of pregnant cows on d 17 of gestation. Nonlactating nonpregnant Holstein cows were assigned randomly to one of 3 treatments: ovulation of a firstwave follicle (FW, n = 15); ovulation of a first-wave follicle and progesterone supplementation (FWP4, n = 12); and ovulation of a second-wave follicle (SW, n = 19). Ovulation of a first-or second-wave follicle was achieved by initiating the Ovsynch protocol (d −9 GnRH, d −2 and −1 PGF 2α , d 0 GnRH and artificial insemination, d 0.7 artificial insemination) on d 0 or 6 of a presynchronized estrous cycle, respectively. Cows in FWP4 received 3 intravaginal inserts containing progesterone at 12, 24, and 48 h after the first GnRH injection that were removed on d −2. Cows were killed on d 17 for collection of the reproductive tract. Transcriptome was evaluated by microarray using the Affymetrix Bovine Array. Orthogonal contrasts were built to assess the effects of progesterone concentration during follicle growth (FW vs. FWP4 + SW) and follicular wave (FWP4 vs. SW). Progesterone concentrations (LSM ± SEM) from d −9 to −2 were greater for SW, followed by FWP4 and FW (5.38 ± 0.24, 4.26 ± 0.28, and 1.17 ± 0.27 ng/mL). Diameter of the ovulatory follicle (FW = 19.6 ± 0.6; FWP4 = 15.6 ± 0.6; SW = 15.2 ± 0.5 mm) and concentrations of estradiol from d −2 to 1 (FW = 4.05 ± 0.33; FWP4 = 2.73 ± 0.35; SW = 2.48 ± 0.30 pg/mL) were greater for FW compared with FWP4 and SW. Progesterone concentrations from d 3 to 16 were greater for FW compared with FWP4 and SW. A total of 28 singleton conceptuses were collected (FW, n = 8; FWP4, n = 8; SW, n = 12) and only intact conceptuses were included in the analyses of length (FW, n = 8; FWP4, n = 6; SW, n = 12). Although conceptuses were longer for FW compared with FWP4 and SW (FW = 16.6 ± 2.3; FWP4 = 9.8 ± 2.2; SW = 9.6 ± 2.0 cm), treatment did not affect the amount of IFN-τ in uterine flushing. Transcriptome of conceptuses and endometrium of pregnant cows was not extensively affected by follicular wave (8 and 1 differentially expressed transcripts) or concentration of progesterone during follicle growth (0 and 3 differentially expressed transcripts), showing that these factors did not affect conceptuses and endometrium transcriptome in pregnancies that are maintained to d 17.

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Validation of an interferon stimulatory response element reporter gene assay for quantifying type I interferons

Cited by 8 publications

References 24 publications

<b>A cell-based interferon-tau assay with an interferon-stimulated gene 15 pro</b><b>moter </b>

<b>A cell-based interferon-tau assay with an interferon-stimulated gene 15 pro</b><b>moter </b>

Exposure to maternal obesity alters gene expression in the preimplantation ovine conceptus

Effects of progesterone concentrations and follicular wave during growth of the ovulatory follicle on conceptus and endometrial transcriptome in dairy cows

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