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Regulated on Activation Normal T Expressed and Secreted Chemokine Is Induced by Tumor Necrosis Factor- in Granulosa Cells from Human Preovulatory Follicle

INSERM, U-355 (V.M., F.N.) and U-131 (D.E.), Institut Paris-Sud sur les Cytokines, 92140 Clamart, France

Address all correspondence and requests for reprints to: Dr. Véronique Machelon, INSERM, U-355, Maturation Gamétique et Fécondation, 32 rue des Carnets, 92140 Clamart, France. veronique. machelon{at}inserm.ipsc.u-psud.fr

We examined the production of regulated on activation normal T expressed and secreted (RANTES) chemokine, which may contribute to the recruitment and local accumulation of leukocytes in human preovulatory follicles. Cells were obtained from follicular aspirates collected from in vitro fertilization patients, then cultured. RANTES production in culture was measured by immunoenzymatic assay, RANTES-producing cells were measured by flow cytometry, and messenger ribonucleic acid as measured by RT-PCR and in situ hybridization. RANTES was detected in follicular fluids and culture supernatants; RANTES protein and messenger ribonucleic acid were expressed in granulosa cells. RANTES production was stimulated by tumor necrosis factor- (TNF) and was inhibited in cultures containing a neutralizing anti-TNF antibody. p55 TNF receptors were detected by RT-PCR and visualized on granulosa cells by flow cytometry. RANTES production was increased by phorbol 12-myristate 13-acetate, but not by 8-bromo-cAMP. RANTES was produced by granulosa cells from human preovulatory follicles. This production was activated by TNF, probably through TNF receptor p55. This suggests that RANTES may play an active role in ovarian processes involving the local accumulation of immune cells.

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