Macrophage and granulosa interleukin-1β mRNA in human ovulatory follicles

Interleukin-1β (IL-1β) 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-1β by granulosa cells and macrophages from the human ovulatory follicle. The concentrations of IL-1β and IL-1β 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-1β was detected by radioimmunoassay and transcripts were detected by RT-PCR and by in-situ hybridization on cytospun preparations using a [ 35 5]IL-1β riboprobe. IL-1β and IL-1β 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-1β transcripts. Moreover, the number of IL-1β 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-1β at ovulation and the ability of granulosa cells to produce IL-1β may require ovarian macrophages.