Author/Editor		Dolinšek-Cencič, Avrelija
Title		Biochemical and functional study of an embryonic interferon-gamma secreted by the porcine trophoblast
Type		monografija
Place		Paris
Publisher		Universite Paris-Sud U.F.R. scientifique d'Orsay
Publication year		2000
Volume		str. 148
Language		eng
Abstract		The pig is so far the only mammalian species in which the trophectoderm of implanting embryo secretes an enormous amount of IFN-gamma (up to 250 p.g/uterine horn). This embryonic IFN-gamma, secreted together with IFN-delta, is physiologieally induced. The peak of its secretion corresponds to the time of embryo implantation. Since this was the first example of a substantial secretion of IFN-gamma by an epithelial cell, and since the function of this "embryonic" IFN-gamma remained uncovered, it was of most interesting to study the structure and function of trIFN-gamma by compariing to those of "adult" leukocytic IFN-gamma. We studied molecular structure and biochemical properties (isoelectric point, pH-stability and ability of refolding, interactions with heparan-sulphate proteoglycans and specific activity in non-endometrial and endometrial cell lines) of epithelia derived trIFN-gamma, and compared them, with those of natural LeIFN-gamma. For some biochemical analyses that demanded large amounts of glycosylated protein to compare with trIFN-gamma, we constructed an inducible, Tc-controlled mammalian expression system to produce recombinant glycosylated IFN-gamma (rGIFN-gamma). The rGIFN-gamma was found to be very similar to the natural LeIFN-gamma, not only in the structure but also in its biological activity. This new rGIFN-gamma was found to be a convenient and economic substitute for the natural IFN-gamma produced by pig leukocytes. An emphasis was given to possible functions of trIFN-gamma and its possible interaction with IFN-gamma. To solve this aim, we used two different approaches: first, we used immunohistochemistry methods on uterine tissue sections to study the IFN-gamma pathway and second, we studied marker differences between the cyclic and pregnant uterus as indices of the embryonic IFN-gamma effects. (Abstract truncated at 2000 characters).
Descriptors		TROPHOBLAST INTERFERON TYPE II PLASMIDS TRANSFECTION CHROMATOGRAPHY, LIQUID CHROMATOGRAPHY, HIGH PRESSURE LIQUID ISOELECTRIC FOCUSING ENZYME-LINKED IMMUNOSORBENT ASSAY SWINE TUMOR CELLS, CULTURED ANTIBODIES, MONOCLONAL