Interleukin-1 in human ovarian cells and in peripheral blood monocytes increases during the luteal phase: evidence for a midcycle surge in the human.

OBJECTIVE: Resident ovarian macrophages are thought to be critically involved in cyclic ovarian events. A prominent macrophage product, interleukin-1, has been shown to affect ovarian cell function. In this report we present evidence for an intrafollicular periovulatory interleukin-1 surge. Additionally, we demonstrate that interleukin-1 beta messenger ribonucleic acid in peripheral blood monocytes increases threefold during the luteal phase of the menstrual cycle over that found in the follicular phase. STUDY DESIGN: Follicular fluid cells isolated as a byproduct of transvaginal oocyte retrievals in gonadotropin-stimulated in vitro fertilization cycles were immunoprobed for the presence of interleukin-1 protein. Late follicular phase cumulus and granulosa cells obtained from an aspirated preovulatory follicle were likewise probed. RESULTS: Although the in vitro fertilization-retrieved cells stained positive for interleukin-1 protein, the late follicular phase cells were devoid of the protein. Granulosa cells from in vitro fertilization cycles were examined for interleukin-1 protein binding sites with iodinated interleukin-1 alpha protein. These cells were found to have approximately 2000 binding sites per cell. Poly A+ messenger ribonucleic acid isolated from peripheral blood monocyte samples from women during the follicular and luteal phases and from male controls were probed for interleukin-1 ribonucleic acid content by means of Northern analysis. The luteal samples contained a threefold higher interleukin-1 messenger ribonucleic acid content that did the follicular phase samples or the controls. CONCLUSION: The ovarian interleukin-1 protein increase taken together with increased peripheral blood monocyte interleukin-1 messenger ribonucleic acid suggests that interleukin-1 production increases at midcycle.

Evidence for the presence of the estrogen receptor in the ovary of the baboon (Papio anubis).

Although intraovarian estrogen has been firmly established as an important factor in the regulation of ovarian follicular development and function in the rat, an autocrine-paracrine role for estrogen in the primate ovary is not yet established. Immunocytochemical identification of an estrogen receptor in the monkey follicle was negative, but it was positive for the granulosa cells of antral follicles of the human ovary. In the present study baboon ovaries obtained during the follicular phase were examined for the presence of estrogen receptor by immunocytochemical analysis of frozen sections and Northern blot analysis of RNA extracts of the ovaries. Immunocytochemistry identified the estrogen receptor in the granulosa cells of healthy appearing and atretic or cystic-like antral follicles and in occasional, but rare, thecal cells. The ovaries contained a prominent mRNA species for the estrogen receptor, approximately 7 kilobases in size, which was present in relatively low abundance compared to that in the nonpregnant baboon uterus, but in a similar abundance to the estrogen receptor mRNA content of the pregnant endometrium. These studies are the first to report the presence of estrogen receptor mRNA in the ovary of a primate. These results in conjunction with the immunocytochemical studies firmly establish the presence of the estrogen receptor in the primate ovary and suggest an autocrine-paracrine role for intraovarian estrogen in primate ovarian physiology.

Ovarian steroids modulate human monocyte tumor necrosis factor alpha messenger ribonucleic acid levels in cultured human peripheral monocytes.

OBJECTIVE: To determine whether tumor necrosis factor alpha (TNF-alpha) messenger ribonucleic acid (mRNA) levels in human peripheral monocytes are regulated by ovarian steroids. DESIGN: Human granulosa-luteal cells and cultured, activated human peripheral monocytes were subjected to Northern blot analysis for TNF-alpha mRNA. SETTING: Academic research laboratory. PATIENTS: Two human female volunteers of reproductive age and in the luteal phase of the menstrual cycle. RESULTS: Human granulosa-luteal cells produce TNF-alpha mRNA. Physiological levels of progesterone (P) and estradiol (E2) modulate TNF-alpha mRNA from peripheral blood monocytes with an apparent inverse relationship between steroid concentration and TNF-alpha message. CONCLUSIONS: Progesterone and E2 at physiological concentrations regulate TNF-alpha mRNA production. The P antagonist mifepristone (RU486) and the E2 agonist/antagonist tamoxifen modulate total TNF-alpha mRNA levels, suggesting involvement of specific receptors.

Production of interleukin 2 and interleukin 4 by immune CD4-CD8+ and their role in the generation of antigen-specific cytotoxic T cells.

In this report we investigated the production and role of interleukin (IL)2 and IL4 in the generation of antigen-specific cytotoxic T cells (CTL). We used as our model the ultraviolet-light-induced epithelial tumor 1591, a highly immunogenic regressor tumor which evokes a strong cell-mediated immune response leading to rejection. We show that IL2 and IL4 are differentially required for the development of optimal cytolytic activity to the 1591 tumor in primary and secondary in vitro splenic cultures. First, anti-IL2 receptor monoclonal antibody (mAb) significantly decreased specific cytotoxicity in both primary and secondary splenic mixed lymphocyte-tumor cell culture (MLTC) cultures, but anti-IL4 mAb inhibited the cytotoxic responses only secondary and not primary cultures. Second, when supernatants from MLTC were tested for lymphokine activity, primary cultures produced only IL2 while secondary cultures produced both IL2 and IL4. Splenic cells were then depleted of CD4+ cells by negative selection, or enriched for CD8+ cells by positive selection, and tested for lymphokine production and requirements. CD8+ cells could not generate significant CTL activity in primary cultures, but could in secondary MLTC. The addition of mAb to either IL2 or IL4 significantly inhibited the generation of CTL by CD8+ cells in these secondary MLTC.CD8+ cells were also found to produce both IL2 and IL4 in secondary MLTC by functional and Northern blot analysis. The production of IL2 and IL4 by CD8+ cells occurs during different phases of culture, with IL2 being produced early (days 1 and 2) and IL4 late (days 3-5). In addition, the requirement of CD8+ cells for both IL2 and IL4 is unique for that lymphokine. These results suggest that both IL2 and IL4 are both produced and required by CD8+ cells during secondary MLTC, and suggest an additional cellular source of IL4 production besides CD4+ T cells during antigen-specific CTL responses.

Human follicular fluids contain tissue macrophages.

Tissue cells obtained from the follicular fluids of women undergoing in vitro fertilization and embryo transfer were found to contain interleukin-1 beta mRNA by Northern analysis. Since macrophages are known to produce interleukin-1 beta, we examined the follicular fluids of 20 women undergoing in vitro fertilization as well as tissue sections of normal human ovary for the presence of macrophages and monocytes. Although granulosa-luteal cells predominate in follicular fluid, we found that resident macrophages and monocytes comprise 5-15% of human follicular tissue cells. In addition, we observed that macrophages are present in the human ovarian follicle as well as in the corpus luteum.