Type

Journal Article

Authors

A D Ealy
P. Lonergan
T E Spencer
P M Mercadante
E B Hall
M Xie
S R McCoski

Subjects

Biochemistry

Topics
cell free interferon ifn interferon type i gene biological samples reporter assay validation bovine cells

Validation of an interferon stimulatory response element reporter gene assay for quantifying type I interferons. (2013)

Abstract The goal of this work was to develop a virus-free, cell-based interferon (IFN) bioassay and determine the utility of this assay on biological samples that contained IFN-τ, the trophoblast-secreted maternal recognition of pregnancy factor in ruminants. Madin-Darby bovine kidney cells were transduced with lentiviral particles that contained a firefly luciferase reporter construct driven by an IFN stimulatory response element (ISRE). Stably transduced cells were selected with the use of puromycin resistance. A linear, dose-responsive response was detected with human IFN-α and ovine IFN-τ. Interferon activity was detected in conditioned media from bovine trophoblast cells and uterine flushes collected from sheep and cattle. Activity also was detected in media collected after individual or small group culture of in vitro-produced bovine blastocysts at day 8 to 10 after fertilization. In summary, this IFN stimulatory response element-reporter assay may be used as an alternative to virus-dependent, cytopathic assays. It contains a similar sensitivity to IFNs and can be completed in a shorter time than cytopathic assays and does not require heightened biosafety conditions after cell transduction.
Collections Ireland -> University College Dublin -> PubMed

Full list of authors on original publication

A D Ealy, P. Lonergan, T E Spencer, P M Mercadante, E B Hall, M Xie, S R McCoski

Experts in our system

1
A D Ealy
University College Dublin
Total Publications: 3
 
2
P. Lonergan
University College Dublin
Total Publications: 190
 
3
Thomas E Spencer
University College Dublin
Total Publications: 14