HUMAN GRANULOSA-CELLS EXPRESS HLA-DR ANTIGEN AND ARE CAPABLE OF SYNTHESIZING INTERLEUKIN-1

To study the origin of interleukin 1 (IL-1) present in human follicula r fluid we determined the percentage of macrophages (MO) and cells wit h HLA-DR antigen (DR+) present in 22 samples of human follicular fluid (FF) from women undergoing in vitro fertilization, and examined the r elease of IL-1 beta by cultures of purified human granulosa cells (GC) . The number of red blood cells (RBC) in the crude preparation was tak en as a measure of possible contamination with peripheral blood monocy tes (assuming a ratio of one monocyte or MO per 10(4) RBC). For the ev aluation of MO and DR+ cells percentages we employed an indirect immun ofluorescence technique using specific monoclonal antibodies. Total ce lls from FF were purified by Ficoll-Hypaque density gradient centrifug ation (delta = 1.076 g/l and GC were purified using a gradient delta = 1.065 g/l. This method reduced the contamination with MO to 0-1 %. Th e spontaneous release of IL-1 beta was measured by ELISA. We found tha t FF contained 9.81 +/- 1.47 % of MO but only 7.85% were ovarian MO. I n addition the total percentage of DR+ cells was 17.13 +/- 2.35% but o nly 9.81% corresponded to MO. Therefore about 7.32% of DR+ cells could be GC. Then purified GC (10(4)/0.2 ml/well) were cultured during 24 h ours at 37 degrees C in serum free medium (DMEM:F12). IL-1 beta levels were 84 +/- 17 pg/ml and these values were increased by 44% when GC w ere stimulated with FSH (200 ng/ml). These results suggest that GC pro duced IL-1 beta and that the synthesis of this cytokine might be regul ated by hormones.