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• W2014467812 abstract "Objective: Regeneration and tolerance factor (RTF) was originally identified in the placenta of mice and the isolated protein shown to have immunosuppressive effects. Thus, the molecule has been suggested to be in part responsible for the survival of fetal allograft. RTF is expressed as a 70 kDa protein which is post-translationally modified by proteolytic cleavage to form a membrane-associated domain of 50 kDa and an extracellular domain of 20 kDa which is a soluble molecule (sRTF). Recently, expression of sRTF has been observed in cytotrophoblast cells and it has been shown to induce the T helper 2 cytokine interleukin (IL)-10 production by immune cells. However, little is known about the expression, function and regulation of the molecule at human endometrial level. Thus, the aims of the present study were to investigate whether RTF mRNA was present in human endometrium and to evaluate the potential modulation of the molecule by IL-1β which is an important cytokine in the endometrial physiology. Design: An experimental study evaluating expression and modulation of RTF in endometrial and decidual samples was performed. Decidua was provided by 25 women undergoing elective termination of normal pregnancies at 8 to 14 wk of gestation. Endometrial samples were obtained from 31 reproductive age women undergoing laparoscopy for benign ovarian cysts or infertility. All the women gave an informed consent. Materials/Methods: Primary cultures were established from all samples of human endometrium and decidua and total RNA was isolated. Expression of RTF mRNA was documented by RT-PCR using two oligonucleotide primers specific for RTF gene. A single major DNA band of the expected size (201 bp) was obtained. The identity of the fragment with the expected RTF sequence was confirmed by both restriction enzyme and sequence analysis. In five different experiments, endometrial cells were treated with and without two different concentrations of IL-1β. Relative RTF mRNA levels were estimated by semiquantitative RT-PCR using hypoxanthine phosphoribosyltransferase 1 gene as internal standard. Products were separated on agarose gels and the bands were densitometrically scanned. A titration series of RT-PCR cycle numbers was performed to verify the linear phase of the amplification. A 25 cycle number was chose to study RTF gene expression. Results: RT-PCR products were consistently detected in all samples of decidual and endometrial cells analyzed. Levels of RTF mRNA were significantly increased in secretory phase when compared to proliferative phase endometrial samples as target/standard ratios were 0.67 ± 0.03 and 0.59 ± .02 respectively (p <0.05). IL-1β was demonstrated to increase endometrial RTF mRNA expression by 7 and 16% respectively at a concentration of 50 and 500 pg/ml. Conclusions: RTF seems to be an important constituent of endometrial system and its up-regulation during the secretory phase of the cycle and by IL-1β supports its involvement in embryo implantation and early phases of pregnancy. Supported by: This project was supported by Istituto Auxologico Italiano." @default.
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• W2014467812 date "2002-09-01" @default.
• W2014467812 modified "2023-10-16" @default.
• W2014467812 title "The gene for regeneration and tolerance factor is expressed by human endometrial cells particularly during the secretory phase of the cycle and is regulated by Interleukin-1β" @default.
• W2014467812 doi "https://doi.org/10.1016/s0015-0282(02)03665-8" @default.
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