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Mediators of Interferon -Initiated Signaling in Bovine Luteal Cells¹

^a The Women's Research Institute, Wichita, Kansas 67214-3199 ^b Department of Obstetrics and Gynecology, University of Kansas School of Medicine, Wichita, Kansas 67214-3199 ^c Veterans Affairs Medical Center, Wichita, Kansas 67218 ^d Vincent Center for Reproductive Biology, Massachusetts General Hospital, Boston, Massachusetts 02114

ABSTRACT

Interferon gamma (IFN) has been implicated as a mediator of luteal steroidogenesis and cell fate. IFN-initiated signaling events, although implied by studies in cell lines, have yet to be described in primary luteal cells. The objective of these studies was to begin to characterize IFN-initiated signaling within luteal cells. Dispersed bovine luteal cell cultures were challenged with increasing levels of bovine recombinant IFN (0–1000 U) or IFN (200 U) in the presence or absence of tumor necrosis factor (TNF, 10 ng/ml) over time (short term, 0–60 min; long term, 0, 24, 48 h). Fractionated or total cell lysates were evaluated by the Western blotting technique to determine the changes in the levels of signal transducers and activators of transcription (STAT), interferon regulatory factor 1 (IRF-1), and I kappa B (IB-). Utilizing antibodies that recognize the nonphosphorylated forms of STAT-1 and STAT-3, it was determined that levels of STAT-1 and STAT-3 in total cell lysates were constitutively expressed and did not change in response to treatment with IFN or TNF. In contrast, nuclear levels of STAT-1 and phosphorylated STAT-3 were elevated in a time-dependent manner in response to IFN treatment. Furthermore, IFN and TNF treatment elevated levels of IRF-1 within 2 h. TNF-induced increases in the levels of IRF-1 were transient, whereas the levels of IRF-1 in response to IFN treatment remained elevated at 48 h. These data suggest that IFN treatment can activate members of the STAT pathway, resulting in increased levels of IRF-1. TNF treatment induced a rapid decrease in the [bu791]levels of IB-. IFN treatment did not alter the levels of IB- and failed to inhibit the TNF-initiated decrease in the levels of IB-. The present experiment demonstrates that the steroidogenic cells of the corpus luteum have the capacity to respond to IFN via activation of STAT and IRF-1, providing further evidence that IFN may be involved in the luteolytic process. These data also suggest that IFN does not signal through the nuclear factor B cell survival signaling pathway.

FOOTNOTES

First decision: 18 October 2000.

¹ Supported in part by NIH grant R01-HD35934 to B.R.R. and J.S.D.

² Correspondence: Bo R. Rueda, Vincent Center for Reproductive Biology, Massachusetts General Hospital, VBK137E-GYN, 55 Fruit St., Boston, MA 02114. FAX: 617 726 7548; brueda{at}partners.org

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