Macrophage and granulosa interleukin-1 beta mRNA in human ovulatory follicles.
Interleukin-1 beta (IL-1 beta) may be active in the ovary at ovulation and may be produced by immune and non-immune cells. This study evaluates the production of IL-1 beta by granulosa cells and macrophages from the human ovulatory follicle. The concentrations of IL-1 beta and IL-1 beta mRNA were measured in follicular aspirates taken from patients undergoing in-vitro fertilization and in cultures of cells from aspirates. Macrophages were detected by immunocytochemistry and by the reverse transcriptase polymerase chain reaction (RT-PCR). The macrophages were removed from granulosa cell preparations immediately or after the cells have been cultured for 24 h. IL-1 beta was detected by radioimmunoassay and transcripts were detected by RT-PCR and by in-situ hybridization on cytospun preparations using a [35S]IL-1 beta riboprobe. IL-1 beta and IL-1 beta mRNA were found in follicular aspirates, in granulosa luteal cell preparations containing macrophages and in highly purified granulosa cell preparations after removal of macrophages by all the methods used. Both macrophages and granulosa cells contained high concentrations of IL-1 beta transcripts. Moreover, the number of IL-1 beta reactive cells in granulosa cell preparations cultured for 24 h with 10-15% macrophages before removal was twice that of granulosa cells cultured without macrophages. Thus, both granulosa cells and macrophages are actively involved in the ovarian production of IL-1 beta at ovulation and the ability of granulosa cells to produce IL-1 beta may require ovarian macrophages.
Machelon
V
V
Institut National de la Santé et de la Recherche Médicale (INSERM), U-355 Maturation Gamétique et Fécondation, Institut Paris-Sud sur les Cytokines, Clamart, France.
Nome
F
F
Durand-Gasselin
I
I
Emilie
D
D
D000328:Adult; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001483:Base Sequence; D002469:Cell Separation; D002478:Cells, Cultured; D005260:Female; D005307:Fertilization in Vitro; D015571:Follicular Fluid / Q000737:chemistry; D006107:Granulosa Cells / Q000737:chemistry*; D006801:Humans; D017403:In Situ Hybridization; D007375:Interleukin-1 / Q000235:genetics*; D008264:Macrophages / Q000737:chemistry*; D008969:Molecular Sequence Data; D006080:Ovarian Follicle / Q000737:chemistry* / Q000166:cytology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis*
D000911:Antibodies, Monoclonal; D007375:Interleukin-1; D012333:RNA, Messenger