We have previously shown that interleukin-8 (IL-8), a cytokine with neutrophil chemotactic/activating and T cell chemotactic activity, is produced by human endometrial stromal and glandular cells in culture. The present study investigated the temporal and spatial expression of IL-8 messenger ribonucleic acid (mRNA) and protein in the human endometrium. Endometrial tissue (n = 52) was obtained fi om human uteri after hysterectomy conducted for reasons other than endometrial disease or from endometrial biopsies. The day of the menstrual cycle was established from women's menstrual history and was confirmed by histology. Half of the tissues (n = 26) were snap-frozen in liquid nitrogen, cellular RNA was extracted, and Northern blots were hybridized with a riboprobe complementary to a specific sequence-of IL-8 mRNA. The remaining tissues (n = 26) were processed for frozen sections, and immunohistochemistry was performed using mouse antihuman IL-8 antibody. Comparison of IL-8 mRNA levels throughout the menstrual cycle revealed that late secretory and early to midproliferative phase IL-8 expression was significantly greater than midcycle expression (P < 0.02). Analysis of the IL-8 immunohistochemistry revealed that IL-8 protein is found in the surface epithelium and glands throughout the menstrual cycle. There was no detectable immunoreactive IL-8 in the stromal cells. We conclude that IL-8 is produced in the human endometrium in vivo, and the variations of IL-8 mRNA throughout the menstrual cycle suggest that sex hormones may regulate its gene expression. We speculate that IL-8 may modulate the timely recruitment of neutrophils and lymphocytes into the endometrium.