H. Oliver Williamson

Interleukin-1: a possible role in the infertility associated with endometriosis *

Increased numbers of activated macrophages are associated with mild endometriosis. Interleukin-1 (IL-1) is a protein produced by macrophages and is believed to be a primary mediator of host responses. IL-1 induces prostaglandin and fibrinogen synthesis and stimulates fibroblast proliferation. This study was undertaken to evaluate the role of IL-1 in the infertility associated with mild endometriosis. Peritoneal fluid (PF) was obtained at laparoscopy from 11 patients with minimal or mild endometriosis and from 7 women undergoing tubal ligation. Peritoneal macrophages were isolated and cultured for 24 hours. Peritoneal and macrophage culture fluids were studied for IL-1 activity, which was measured with the EL-4 assay. IL-1 activity was present in the PF of 10 of the 11 patients with endometriosis and 11 of the 11 macrophage culture fluids and was absent in the PF and macrophage culture fluid of the tubal ligation patients. The effect of recombinant alpha-IL-1 on the in vitro growth of 2-cell mouse embryos was also studied. IL-1 in concentrations similar to those present in the PF (greater than 1 U/ml) was toxic to mouse embryo development. We conclude that the IL-1 may play a role in the infertility associated with endometriosis.

Female runners and secondary amenorrhea: correlation with age, parity, mileage, and plasma hormonal and sex-hormone-binding globulin concentrations*†

Twenty-three white women, ages 18 to 42, with normal menses prior to running were studied. Miles per week varied from 10 to 70 for a period of 1 to more than 10 years. Of these, 6 were amenorrheic (AM), 14 had regular cycles (REG), and 3 with regular cycles became amenorrheic during the course of this study. The incidence of amenorrhea was higher in those less than 30 years of age (66.6%) than in the older group (9.0%); in those who ran 40 miles/week or less (37.5%) than in those who ran more (26.6%); and in the nulliparous (46.6%) than in the parous runners (25.0%). The age of menarche was significantly higher in the AM (13.8 +/- 0.5 years) than in the REG (12.2 +/- 0.3 years). Blood samples were collected between 12 and 24 hours after the last run for hormonal and sex-hormone-binding globulin (SHBG) measurements. Plasma estradiol (E2), SHBG, and LH were significantly lower in the AM than in the REG group. Furthermore, E2, LH, and prolactin were significantly lower in the AM group than in the control group. These results suggest that the incidence of secondary amenorrhea is higher in younger, nulliparous female runners and may be related to delayed onset of menarche.

Plasma Androgens and Sex Hormone-Binding Globulin in the Evaluation of Hirsute Females *

Hirsutism is usually associated with increased testosterone (T) production and metabolic clearance rates. Considerable overlap of plasma T occurs between hirsute and normal groups. Plasma levels of sex hormone-binding globulin (SHBG) and the factor T/SHBG might separate hirsute patients from normal subjects better than plasma T. A group of 39 hirsute females and 22 normal ovulatory control subjects were studied. Plasma T, androstenedione, and dehydroepiandrosterone were measured by radioimmunoassay; apparent free T (AFT) by equilibrium dialysis; and SHGBG by a method based on saturating the binding sites by labeled dihydrotestosterone. Mean levels of androgens and SHBG of the hirsute patients were significantly different from those of the normal subjects (P less than 0.01). Positive linear correlations were observed between T and AFT, T/SHBG and AFT, and T/SHBG and T; a negative correlation was observed between T/SHBG and SHBG, but no correlation was observed between SHBG and T or AFT. Thirty (77%) of the patients had elevated T/SHBG factors and 28 (72%) had suppressed SHBG levels. Only two patients (5%) had hirsutism associated with normal levels of androgens, SHBG, and T/SHBG. We conclude that SHBG and the factor T/SHBG separate the hirsute population better than any of the androgens studied.

Cyclic variations in white cell subpopulations in the human menstrual cycle: Correlations with progesterone and estradiol

Cyclic variations in white cell subpopulations were studied in serial blood samples from 18 female volunteers (14 ovulatory and 4 nonovulatory cycles) and 2 males. Total white blood cells (WBC), lymphocytes, total and active T cells (TEt, TEa), monocytes, and granulocytes were counted, and levels of estradiol (E2), progesterone, and luteinizing hormone (LH) were measured. In the ovulatory cycles, lymphocyte counts at midcycle (Day 0) reached a minimum, coinciding with the maximum level of E2 peak (35 ng%). In daily samples, the minimum lymphocyte counts coincided with the preovulatory E2 surge (P < 0.01). Similar but less significant negative correlations (P < 0.05) were found between E2, WBC, and TEt. In contrast, TEa did not show cyclic variations. Monocyte and granulocyte counts were significantly higher in the luteal than in the follicular phase (P < 0.05); their pattern followed closely that of progesterone (P < 0.05) but not of E2. Daily evaluations of hematocrit, hemoglobin content, red blood cell count, and mean corpuscular volume failed to show cyclic variations. For the females with nonovulatory cycles and for the males studied (with sex steroid profiles corresponding to the follicular phase), no cyclic variations were found in white cell subpopulations.

The effect of estrogen treatment on plasma concentrations of steroid hormones, gonadotropins, prolactin and sex hormone-binding globulin in post-menopausal women

Twenty-one post-menopausal women on no other medication were treated with a low dose (0.625 mg/day) of conjugated equine estrogen (CEE) for a mean (+/- SEM) period of 2.6 +/- 0.2 mth (range 1.75-4.75). Blood samples were collected before and at the completion of therapy, and alterations in the levels of prolactin (PRL), follicle-stimulating hormone (FSH), luteinizing hormone (LH), sex hormone-binding globulin (SHBG) and certain steroid hormones, including the free testosterone (T) index (T/SHBG) were studied. Following treatment, a significant increase in SHBG levels produced a significant decrease in the free T index (P less than 0.005). As expected, no changes were observed in the levels of PRL and steroid hormones other than estrone (E1) and estradiol-17-beta (E2). Our observations indicate that treatment of post-menopausal women with low-dose estrogen lowers the unbound T.

Clinical significance of sperm antibodies in infertility

Sperm antibody (AB) titers, determined by passive hemagglutination and cytotoxicity assays, were found to be elevated in 62 males and 46 females of 103 couples with primary infertility; 15 males and 12 females of 25 couples with secondary infertility; 10 males and 8 females of 18 couples with histories of repeated abortion; 21 males and 17 females of 25 couples in which the husband had a history of prostatitis; and 29 males and 17 females of 38 couples in which the husband had oligospermia. Of the couples in which one or both partners had elevated sperm AB titers, only 4 achieved pregnancy: 3 from the group with secondary infertility, all of which ended in spontaneous abortions, and 1 in which the husband was oligospermic. This suggests an etiologic role of sperm immunity in infertility. Immunosuppressive treatment of autoimmune males with prednisone (15 mg/day for 3 weeks to 6 months) resulted in significant decreases in AB titers. Pregnancies were achieved by 9 of 25 couples after treatment (36%). The observed increase in pregnancy rate in the prednisone-treated versus untreated groups of couples with elevated sperm AB titers was significant (P less than 0.02).

Endometrial antigens involved in the autoimmunity of endometriosis.

Serum and peritoneal fluid from five fertile women without endometriosis and serum (n = 23) and peritoneal fluid (n = 12) from infertile women with endometriosis were tested for the presence of antibodies against endometrial tissue antigens by a Western blot analysis. Antigens with molecular weights (MW) of 19, 31, 38, and 42 kd reacted with antibodies in the serum and peritoneal fluid from both fertile and infertile women. Antibodies in 20 of 23 (87%) sera and all 12 (100%) peritoneal fluid samples from endometriosis patients reacted against endometrial antigens with molecular weights (MW) of 26 kd and/or 34 kd. Serum from 10 patients (43%) and peritoneal fluid from 6 patients (50%) also had antibodies to an endometrial antigen with MW of 21.5 kd. Reactivity to other endometrial antigens with MW 16, 24, 48, and 75 kd was also noted in patients with endometriosis. Antibodies in the serum and peritoneal fluid from fertile women failed to react against these antigens. It is concluded that the humoral and local endometrial autoimmunity detected in patients with endometriosis is primarily directed against antigens with MW of 26 and 34 kd.

Plasma gonadotropins, prolactin, and steroid hormone concentrations in female runners immediately after a long-distance run

Six normally menstruating women who regularly run participated in a 10-mile race. Blood samples were collected within 20 minutes after the completion of the race (group 1). Samples were analyzed, and the results were compared with plasma hormonal concentrations in the same runners in samples collected between 12 and 24 hours after a previous practice run (group 2) and with our nonathletic female controls. Plasma concentrations of the following hormones in group 1 were significantly elevated when compared with group 2: dehydroepiandrosterone (DHA), androstenedione (delta 4A), testosterone (T), cortisol (F), luteinizing hormone (LH), and prolactin (PRL). Levels of 17 beta-estradiol (E2), 17-hydroxyprogesterone (17-OHP), dehydroepiandrosterone sulfate (DHAS), and follicle-stimulating hormone (FSH) were comparable in groups 1 and 2. However, DHAS in group 1 was elevated when compared with controls, as were HDA, delta 4A,T,F,LH, and PRL. In group 1, but not in group 2, a significant correlation (P less than 0.05) was observed between plasma LH and PRL concentrations but not between FSH an PRL. We conclude that the immediate effect of running is reflected in increased levels of the adrenal androgens, F, LH, and PRL. However, concentrations of these hormones revert back to baseline within 12 to 24 hours after the race.

An endometrial antibody assay in the clinical diagnosis and management of endometriosis

Coded serum samples from 11 normal fertile men and 17 fertile women without endometriosis (control groups) and 41 women with endometriosis were tested blindly for the presence of endometrial antibodies by use of a passive hemagglutination assay. Endometrial antibodies were either absent or present in low baseline titers in the serum samples from the control group. In contrast, 17 of the 23 (74%) patients with untreated endometriosis had elevated titers of endometrial antibodies in their serum. Of the 18 patients treated with danazol, endometrial antibodies were absent in 7 women who showed a good response at repeat laparoscopy, whereas 4 of 5 women with a poor response had significantly positive titers of antibodies. Six patients treated with danazol did not have repeat laparoscopy, but were found to have endometrial antibody titers in the baseline control range. Endometrial antibody titers in women with a good response to danazol were significantly lower than those in women with untreated endometriosis or with a poor response to danazol (P = 0.003). No correlation was observed between the antibody titers and the stage of endometriosis. The results suggest that the assay for serum endometrial antibodies may prove to be a clinically useful, noninvasive aid in the diagnosis of endometriosis. Sequential determination of endometrial antibody titers may be helpful in assessing the efficacy of pharmacologic therapy for endometriosis and evaluating the cases of patients with possible recurrence of the disease.

Cross-reactivity of sperm and T lymphocyte antigens.

ABSTRACT: Evidence is presented for cross‐reactivity between antigens on human sperm and T lymphocytes. In 25 infertile couples in which both the males and females had significant antisperm immunity, antibody (Ab) titers to thymocytes (mean ± S.E.M. 159 ±4 and 72 ± 14, respectively, in males and females), T cell lines CCRF‐CEM (69±5 and 48±8) and HSB‐2 (56±15) and 41±8), suppressorenriched (TG) cells (26±6 and 66±28) and helper‐enriched (Tg—) cells (26±4 and 46±14) were significantly elevated, as compared wth Ab titers in 45 normal males and 45 normal females without antisperm immunity. Antibody titers to adult B cells, B cell line RAJI, and granulocytes were similar in the two groups. Antisperm Ab titers in sera, sperm extracts, and seminal plasma of the infertile subjects were significantly reduced after absorption with sperm, thymocytes, or T cell line CCRF‐CEM but not with the B cell line RAJI. Antithymocyte Ab titers in the sera were significantly reduced (p < 0.001) after absorption with thymocytes, CCRF‐CEM, or sperm, but not RAJI. Lymphocytes from the infertile patients, when stimulated with pokeweed mitogen in vitro, produced antisperm and anti‐T‐lymphocyte antibodies at significantly higher titers than normal controls.