Activation status of T and NK cells in the endometrium throughout menstrual cycle and normal and abnormal early pregnancy.

Endometrial lymphocytes were studied at all stages throughout the menstrual cycle and early pregnancy by flow cytometry to examine different lymphocyte subpopulations and the expression of the T- and NK-cell activation markers. After pregnancy, CD8+CD3+ lymphocytes were decreased in the decidua. In both endometrium and decidua, more T cells expressed CD69, CD71, HLA-DR, and CD38 antigens than in peripheral blood. After pregnancy, CD71+CD3+ lymphocytes were further increased. CD25+CD3+ lymphocytes decreased significantly in the endometrium and decidua of ectopic pregnancies, but not in the decidua of normal pregnancies. These findings indicate that T cells are regionally activated in the first trimester, and it may be the result of the stimulation by fetal antigens. NK cells were the most abundant cell type in the decidua, which expressed the phenotype CD16- CD56+, and CD57-CD56+. The proportion of activated decidual NK cells was increased in anembryonic pregnancies more than in normal pregnancies, although the total NK subpopulation was similar in both groups. This might result in increased NK cytotoxicity in anembryonic pregnancies. In conclusion, T cells are activated, but NK cytotoxicity is decreased in the decidua of early normal pregnancies. This might be important in the control of trophoblast growth and placental development.

The expression of killer cell inhibitory receptors on natural killer cells and activation status of CD4+ and CD8+ T cells in the decidua of normal and abnormal early pregnancies.

The establishment of the human placenta in early pregnancy is characterized by the presence of large numbers of natural killer cells within the maternal decidua. These NK cells have an unusual phenotype, CD3- CD16- CD56(bright), distinguishing them from peripheral blood NK cells. They may control trophoblast migration and placentation. Using a panel of monoclonal antibodies to several members of the KIR family and flow cytometry, we found that KIRs are expressed on decidual NK cells. There is variation in both the percentage of cells expressing a particular receptor and the density of receptor expression between decidual NK cells from different individuals. In anembryonic pregnancy, the proportions of decidual NK cells with a particular KIRs (GL183 and EB6) decreased significantly when compared with normal pregnancy (p = 0.01 and 0.01, respectively), raising the possibility that these NK receptors may be involved in recognition of the allogeneic fetus by the mother at the implantation site. In the decidua, more CD4+ and CD8+ T cells expressed CD69 and HLA-DR than in blood, indicating that T cells are regionally activated during early pregnancy. When compared with normal pregnancy, decidual HLA-DR+CD4+CD3+, CD69+CD8+CD3+ and HLA-DR+CD8+CD3+ T lymphocytes are significantly increased in anembryonic pregnancy. The over-activation of decidual T cells during anembryonic pregnancy may thus contribute to the increased NK cytotoxicity activity.

Effects of cryopreservation on meiotic spindles of oocytes and its dynamics after thawing: clinical implications in oocyte freezing--a review article.

Embryo freezing has been a successful practice, but oocyte cryopreservation formerly achieved poorer results. This was mainly due to low rates of survival, fertilization, and development. The major dissimilarities for oocytes to embryos are the character of the plasma membrane, the presence of cortical granules, at the metaphase of meiosis II with the spindle system. In addition, the oocytes must be fertilized by sperm at the appropriate time. To improve the survival rate, a refined slow freezing method with increased sucrose concentration would dehydrate oocytes more sufficiently. Vitrification is another approach to prevent ice crystal formation. Intracytoplasmic sperm injection is used to overcome possible zona hardening from the release of cortical granules. The microtubules of meiotic spindles are vulnerable to the thermal changes and would depolymerize. Cryopreserved oocytes exhibited serious disturbances of the microtubules immediately after thawing. Fertilization of oocytes with disorganized spindles could lead to chromosomal aneuploidy, digyny, and arrest of cleavage. After incubation, the microtubules would repolymerize in a time-dependent way. Normal fertilization and development of cryopreserved oocytes improved after appropriate incubation and timing of insemination, compatible with recovery of the spindles. With the improvement of survival, fertilization, and cleavage, oocyte cryopreservation would gain an imperative role.

Hormone replacement therapy reverses the decrease in natural killer cytotoxicity but does not reverse the decreases in the T-cell subpopulation or interferon-gamma production in postmenopausal women.

OBJECTIVE: To investigate the immunologic deviations of postmenopausal women before and after hormone replacement therapy (HRT). DESIGN: Prospective study. SETTING: University teaching hospital. PATIENT(S): Seventeen postmenopausal women (study group) and 17 women of reproductive age (control group). INTERVENTION(S): Continuous usage of E(2) valerate 2 mg/d and medroxyprogesterone acetate 5 mg/d in postmenopausal women in the study group. MAIN OUTCOME MEASURE(S): Immunophenotyping with flow cytometry, cytokine production with and without mitogen stimulation of the peripheral mononuclear cells, and a natural killer (NK) cell cytotoxicity test against K562 target cells by the (51)Cr-release assay were performed in the control group and in the study group before, 1 month after, and 6 months after HRT. RESULT(S): NK cytotoxicity, interferon-gamma production, and the T-cell subpopulation were significantly decreased, and the subpopulations of CD3(+)CD25(+) and CD3(+)HLA-DR(+) were increased in the study group before HRT when compared with those in the control group. After HRT was given for 6 months, however, the NK cytotoxicity increased significantly in the postmenopausal women to a value similar to that of the control group. CONCLUSION(S): Women after menopause are prone to impaired immune responses. Nevertheless, some of the impairment can be restored after HRT.

Increase in the expression of killer cell inhibitory receptors on peritoneal natural killer cells in women with endometriosis.

OBJECTIVE: Malfunction of peritoneal natural killer cells (NK) may result in endometriosis. The present study was designed to determine whether the decrease in NK cytotoxicity occurs at early and advanced stages of endometriosis and is due to the increase in the NK inhibition receptors. DESIGN: A case control study. SETTING: A tertiary-care infertility center . PATIENT(S): A total of 44 women (controls, n = 11; women with early-stage endometriosis, n = 11; and women with advanced-stage endometriosis, n = 22) were included in this study. INTERVENTION(S): Laparoscopic examination. MAIN OUTCOME MEASURE(S): NK cytotoxicity was determined by assay of (51)Cr release against K562 cells, and the expression of killer cell inhibitory receptors (KIR, including NKB1, GL183, and EB6) in NK cells was examined by flow cytometry. RESULT(S): Women with endometriosis showed a decrease in peritoneal NK cytotoxicities against K562 at early and advanced stages of endometriosis. The expression of KIR (NKB1 and EB6) was significantly elevated in the peritoneal NK cells of women with advanced-stage endometriosis compared with controls. KIR (NKB1) was also significantly increased in peritoneal NK cells of women with advanced-stage endometriosis, compared with those of women with early-stage endometriosis. CONCLUSION(S): The results of this study suggest that the decrease in peritoneal NK cytotoxicities against K562 is observed and that this disease may be partially due to the increased expression of KIR on these NK cells.

The simplified two-pipette technique is more efficient than the conventional three-pipette method for blastomere biopsy in human embryos.

OBJECTIVE: To evaluate the efficiency and efficacy of a simplified two-pipette technique in comparison to the conventional three-pipette method; in the two-pipette method, a single, larger drilling/biopsy pipette is used to perform zona pellucida (ZP) drilling and blastomere aspiration for embryo biopsy. DESIGN: A preclinical, prospective, randomized, in vitro experiment. SETTING: The reproductive unit of a university teaching hospital. PATIENT(S): Ninety-five excess embryos at the two- to four-cell stage were obtained from 35 patients undergoing IVF. INTERVENTION(S): At the six- to eight-cell stage, 88 embryos were allocated randomly to three groups: group I for the conventional method (n = 29), group II for the simplified technique (n = 30), and group III for controls (n = 29). The embryos then were cultured in vitro. The retrieved blastomeres were fixed and examined with fluorescence in situ hybridization using X and Y probes simultaneously. MAIN OUTCOME MEASURE(S): Biopsy time, successful retrieval of a blastomere, fixation of the cell, signals developed from fluorescence in situ hybridization, and growth potential and hatching capacity of the biopsied embryos were evaluated. RESULT(S): The mean time (+/- SD) for biopsy of each embryo in group I (435 +/- 137 seconds) was significantly longer than that in group II (126 +/- 32 seconds). The success rates for obtaining an intact blastomere were not different between group I (93%) and group II (97%). The growth capacity to the blastocyst stage was similar among the three groups (34%, 37%, and 38%, respectively). However, the ZP-drilled and biopsied embryos of groups I and II had higher percentages of hatching (34% and 37%, respectively) and complete hatching (17% and 20%, respectively) than did those of group III (10% and 0, respectively). The blastomeres obtained by biopsy in groups I and II were equally fixed (90% vs. 90%, respectively) and shown in fluorescence in situ hybridization (79% vs. 80%, respectively). CONCLUSION(S): Compared with the conventional method, the simplified technique is more efficient and equally efficacious for blastomere biopsy in preimplantation genetic diagnosis.

Distribution of Th1 and Th2 cell populations in human peripheral and decidual T cells from normal and anembryonic pregnancies.

OBJECTIVE: To examine whether maternal immune responses during normal pregnancy are Th2 biased and whether there are specific changes when anembryonic pregnancy occurs. DESIGN: Prospective study. SETTING: Department of Obstetrics and Gynecology at a university hospital. PATIENT(S): We studied 32 pregnant women receiving elective abortions of normal pregnancies and 35 women with anembryonic pregnancies between 6 weeks and 10 weeks of gestational age. INTERVENTION(S): Using the multilabeling capability of three-color flow cytometry, it is possible to measure intracellular cytokines and cell surface markers simultaneously to determine which cells are the cytokine-producing cells. MAIN OUTCOME MEASURE(S): We examined the extent and proportion of mononuclear cells expressing specific T-cell surface markers and cytokines, interferon gamma, and interleukin 4 in the peripheral blood and deciduae. Secreted cytokines in the supernatants after 24-hour culture were also compared. RESULT(S): During the unstimulated status, the proportion of IL-4-secreting cells significantly exceeded that of IFN-gamma-secreting cells in the peripheral blood and decidua in normal pregnancies and was significantly decreased when anembryonic pregnancies occurred. Consequently, the Th1/Th2 ratios were increased during anembryonic pregnancies. However, after 24-hour culture, only another Th2-type cytokine, IL-10, was markedly increased and exceeded IFN-gamma secretion in cultures from both the peripheral blood and decidua in normal pregnancies. CONCLUSION(S): The decidual T lymphocytes are Th2 predominant. When anembryonic pregnancy occurs, this Th2 predominance disappears.

Assisted hatching increases the implantation and pregnancy rate of in vitro fertilization (IVF)-embryo transfer (ET), but not that of IVF-tubal ET in patients with repeated IVF failures.

OBJECTIVE: To assess the effect of augmenting IVF with assisted hatching in the treatment of patients with repeated IVF failures. DESIGN: Prospective randomized study. SETTING: Division of Reproductive Endocrinology and Infertility of National Taiwan University Hospital. PATIENT(S): From July 1993 to February 1996, 49 patients with repeatedly failed IVF were treated with assisted hatching and were compared with 51 control subjects without assisted hatching. INTERVENTION(S): Assisted hatching. MAIN OUTCOME MEASURE(S): Pregnancy rate and implantation rate per embryo after IVF-ET or IVF-tubal ET (TET) were measured. RESULT(S): The pregnancy rate (PR) in the assisted hatching group was found to be 36.7% compared with 19.6% in the control group, but the difference was not significant. When only patients receiving IVF-ET were considered, it was observed that the PR was significantly higher in the assisted hatching group than the control group (42.4% versus 16.1%). With IVF-TET however, the PR was found to be similar in both assisted hatching and control groups (25.0% and 25.0%, respectively). The rate of embryonic implantation in the IVF-ET patients was 11.0%, which was significantly higher than that of control embryos (3.7%). CONCLUSION(S): These results implied that IVF-ET, combined with assisted hatching, may improve the PR and implantation rate in patients with repeated IVF failures, but the same was not true in the case of IVF-TET.

Histopathologic characteristics of a chronic arteriovenous malformation in a swine model: preliminary study.

BACKGROUND AND PURPOSE: The experimental induction of histologic transformations in microvessels of similar caliber to those of nidus vessels of cerebral arteriovenous malformations (AVMs) has not been attempted previously. Our goal was to examine preliminarily the histopathologic characteristics of nidus vessels and the angiographic features of a chronic AVM model in swine. METHODS: AVM models were fashioned from bilateral carotid retia mirabilia of seven swine after the surgical formation of large unilateral carotid-jugular fistulas. One AVM model was made for immediate use, whereas in the other six, follow-up angiography was obtained at varying intervals (2 to 180 days) after model creation. Light and electron microscopy, immunohistochemistry (using monoclonal antibodies against smooth muscle actin and PC10 against proliferating cell nuclear antigen), and histometry were performed on the nidus vessels of three swine: one acutely created, one 2 months old, and one 6 months old. RESULTS: Vascular dilatation and tortuosity of the main arterial feeder and draining vein were evident angiographically as early as 4 days after AVM creation, and were maximal in the 6-month-old model. Compared with the acutely created nidus vessels, those in the two chronic models revealed disrupted and attenuated elastica and intimal hyperplasia that was focal ("cushions") or generalized, leading to luminal occlusion. Variable numbers of cells in the tunica media of chronic nidus vessels contained smooth muscle actin. PC10/proliferating cell nuclear antigen immunoreactivity was observed in the endothelium and subendothelial layers. Histometry showed increases in intimal hyperplasia and medial thickness in the chronic vessels. CONCLUSION: Nidus vessels in this chronic swine AVM model exhibited striking histologic changes similar to those seen in cerebral AVMs. The induced vessel growth seen angiographically and histologically in components of the chronic AVMs was consistent with the presence of persistently raised intravascular hemodynamic loads. This preliminary feasibility study suggests that the realistic histologic characteristics of this chronic AVM model are an attractive feature, and if confirmed in future, more comprehensive, studies would be of benefit in accurate histopathologic interpretation of the effects of superimposed experimental embolotherapy or radiosurgery. This model may provide a useful experimental tool to study the dynamic cellular and tissue events that dictate the development and natural history of AVMs.

KTP laser and neutral red phototherapy of human squamous cell carcinoma.

Neutral red (NR) is a cationic, nontoxic vital dye employed as a histologic stain for proliferating cells; it has been used clinically for photodynamic treatment of herpes simplex virus lesions. NR is selectively taken up and concentrated by mitotic cells, an important characteristic for more effective antineoplastic agents. In the present study, UCLA-SO-P3 human squamous carcinoma cells displayed minimal toxicity when incubated with up to 50 microg/ml NR in the absence of light. However, cells incubated with greater than 0.5 microg/ml NR followed by exposure to KTP laser light at 532 nm exhibited nearly 100% tumor cell death. The degree of cell toxicity was proportional to NR dose and laser light fluence. This study demonstrates that NR is an excellent cancer cell photosensitizer in vitro, and, after adding additional in vivo preclinical testing, may prove to be a useful agent in photodynamic destruction of head and neck tumors.

Synchronous primary carcinomas of the endometrium and ovary.

OBJECTIVES: Synchronous carcinomas of the endometrium and ovary may indicate either independently developing neoplasms or metastatic disease. The clinical implications and prognosis of these two categories are quite different. The objectives of this study were to identify and evaluate the empirical criteria and significant therapeutic implications. METHOD: The National Taiwan University Hospital Cancer Registry records and pathological reports from 1977 to 1994 were reviewed. Empirical criteria were used to identify synchronous primary cancers. RESULTS: A total of 322 patients had endometrial cancer and 421 patients had ovarian cancer in our Cancer Registry records. Eleven patients had simultaneous cancer involvement of both the endometrium and ovary. Six cases fulfilled the criteria of synchronous primary carcinomas of the endometrium and ovary. Of these, five were alive and free of disease for 35-144 months (median 94.2 months). The disease-free survival rates between patients with synchronous primary and metastatic cancers of different histologic types showed a statistically significant difference (P = 0.013). No statistical significance was noted for different histologic types (P > 0.5). CONCLUSIONS: The empirical criteria used here were useful in identifying synchronous primary cancers of the endometrium and ovary. The favorable clinical outcome may relate to early detection of early-stage disease and low-grade malignancy with an indolent growth rate. Surgical management with or without adjuvant therapy has a satisfactory outcome in our experience.

Long GnRH-agonist protocol in an IVF program. Is it appropriate for women with normal FSH levels and high FSH/LH ratios?

OBJECTIVE: To determine whether subjects with an elevated ratio of follicle-stimulating hormone (FSH) to luteinizing hormone (LH) but normal basal FSH levels should be regarded as poor responders to controlled ovarian hyperstimulation. STUDY DESIGN: One hundred twenty-five women undergoing in vitro fertilization (IVF) for the first time were recruited in this retrospective cohort study. Women over 40 years old or having serum basal FSH > 10 mIU/mL were excluded. RESULTS: Various cutoff values for the FSH/LH ratio were chosen, and the ratio demonstrated that pregnancy rates were apparently higher in patients with the long protocol than with the short one if they had an FSH/LH ratio < 3.0 (48.5% vs. 25.8%, P = .034), < 2.5 (53.3% vs. 28.6%, P = .030) or < 2.0 (57.8% vs. 21.7%, P = .005). Pregnancy rates were similar with the long and short protocols in patients with FSH/LH > or = 3.0 (57.1% vs. 70%, P = .521), FSH/LH > or = 2.5 (40% vs. 53.8%, P = .435) or FSH/LH > or = 2.0 (40% vs. 55.6%, P = .281). CONCLUSION: This study failed to demonstrate that FSH/LH was a useful parameter for predicting reproductive outcome in IVF programs and for patient selection for the long or short gonadotropin-releasing hormone agonist protocol.

Controlled ovarian hyperstimulation and intrauterine insemination in subfertility. How many treatment cycles are sufficient?

OBJECTIVE: To examine the combined effects of factors influencing the pregnancy rates following controlled ovarian hyperstimulation (COH) and intrauterine insemination (IUI). STUDY DESIGN: After utilizing the couple as the unit of analysis, 416 couples undergoing COH and IUI with the husband's sperm were studied retrospectively. RESULTS: The woman's age, postwash total motile sperm counts and number of IUI treatment cycles were three major factors that affected the results of IUI. The predicted cumulative pregnancy rates under different values of the above three factors were therefore calculated using multiple logistic regression analysis and were compared to the pregnancy rate achieved by a cycle of in vitro fertilization and tubal embryo transfer. CONCLUSION: Three treatment cycles of IUI are optimal for women aged < or = 34 with postwash total motile sperm counts > 5 x 10(6). For women either aged > or = 35 or with postwash total motile sperm counts < 5 x 10(6), the cumulative pregnancy rates achieved by IUI are extremely low, and other means of assisted reproduction are recommended for couples in those categories. However, this conclusion pertains to our clinics, and other institutions should establish their own data.

Cornual pregnancy after IVF-ET. A report of three cases.

BACKGROUND: Ectopic pregnancy is a well-known complication of in vitro fertilization/embryo transfer (IVF-ET). Although ectopic pregnancy after IVF-ET in patients with prior bilateral salpingectomy is uncommon, it can occur and may be a cornual implantation. Because of the possibility of early rupture with severe hemorrhage, special attention to cornual pregnancy after IVF-ET is warranted. CASES: Three cases of cornual pregnancy occurred after IVF-ET. Two of these patients had prior bilateral salpingectomy, whereas another had prior tuboplasty for tubal disease. CONCLUSION: Patients who had prior salpingectomy or tuboplasty with cornual patency undergoing IVF-ET are at risk for cornual pregnancy. Close monitoring of pregnancies in these patients is important to prevent a deleterious delay in treatment of a cornual pregnancy.

Methotrexate treatment for cervical pregnancy: experience of four cases.

Although cervical pregnancies account for a very small number of the ectopic pregnancies, they sometimes result in a tragic outcome due to the difficulty of surgical treatment and subsequent intractable hemorrhage. We report our experience with four cases of cervical pregnancy which were treated successfully with methotrexate. All four patients had a history of a previous abortion and two had previously had a cesarean section. High-resolution transabdominal and transvaginal color Doppler ultrasound was used for diagnosis, and the serial beta-human chorionic gonadotropin levels were checked to monitor the therapeutic effect. Hysterectomy was avoided, and the reproductive capability of these patients was preserved. Chemotherapy was well tolerated except for mild stomatitis in one case and a transient elevation of liver enzymes in another case. One patient delivered a healthy female infant two years after treatment. In conclusion, previous curettage and a low segment cesarean section may contribute to the development of a cervical pregnancy. Fortunately, methotrexate therapy offers an effective nonsurgical treatment for this potentially catastrophic condition.

Heterotopic pregnancies after controlled ovarian hyperstimulation and assisted reproductive techniques.

Eight cases of heterotopic pregnancy following assisted reproductive techniques (ART) were reviewed. One cervical, three cornual and four tubal pregnancies were combined with intrauterine pregnancies. The cervical ectopy was successfully treated with a local injection of potassium chloride (KCl) into the gestational sac. A viable baby was produced from the intrauterine gestation. Cornual metroplasties were performed on two ruptured and one unruptured cornual heterotopic pregnancies. One of these three cases was maintained till term. All four tubal heterotopic pregnancies were aborted. Widespread application of ART in recent years has meant that heterotopic pregnancies are no longer a rarity. Both physicians and patients should be made aware that the existence of an intrauterine gestation does not preclude the risk of nidation of other fetuses in ectopic sites. The authors recommend that detailed ultrasound studies, preferably via the vaginal route, should be performed on patients in ART programs. During the examination, the adnexae should be carefully evaluated, even if an intrauterine gestational sac is already present. If a heterotopic pregnancy is diagnosed, the appropriate treatment depends on the location of ectopic pregnancy. Local injection with KCl or methotrexate is effective in certain types of cervical or cornual ectopic pregnancy, as it may allow the conservation of the intrauterine pregnancy.

Effect of assisted hatching by partial zona pellucida dissection on mouse embryos in vitro.

The zonae pellucida of mouse embryos were dissected to examine the effect of partial zona dissection (PZD) at the four-cell stage on embryonic development and the hatching process in vitro. Micropipettes were used to create a gap of one-sixth the zona circumference, to avoid damage to the blastomere. A total of 484 embryos at the four-cell stage (defined as day 2 embryos) were randomly divided into two groups, the PZD group (n = 238) and the control group (n = 246). Embryos of both groups were cultured in vitro under identical conditions and examined on the following day. The hatching rates were as follows: day 3, 11/238 (4.6%) in the PZD group and 0/246 (0%) in the control group; day 4, 105/238 (44.1%) in the PZD group and 54/246 (22.0%) in the control group; day 5, 71/238 (29.8%) in the PZD group and 36/245 (14.5%) in the control group. On day 4 the rates of complete hatching were 7/238 (2.9%) in the PZD group and 2/246 (0.8%) in the control group. On day 6 they were 88/238 (37.0%) in the PZD group and 66/246 (26.8%) in the control group. A significantly higher percentage of PZD embryos than controls initiated and completed hatching in similar culture conditions. The present study indicates that the creation of an opening in the zona pellucida is not harmful to embryonic development and may facilitate hatching. Whether the enhanced hatching ability of embryos in vitro caused by PZD can improve the implantation rate requires further study.

Vulvar neoplasia associated with other primary malignancies.

It is recognized that a significant number of patients with vulvar neoplasia have had previous, concurrent, or subsequent genital tract neoplasia. This has resulted in speculation that there may be a common etiology and, in particular, of the possibility of an infection element. Among 59 cases of vulvar cancer diagnosed at the National Taiwan University Hospital from 1976 to 1991, there were seven cases (12%) associated with other primary malignancies. Carcinoma of the cervix was the most frequent other primary cancer (6/7, 86%). The possible role of human papilloma virus infection in genital neoplasia is discussed.

Serum estradiol level and oocyte number in predicting severe ovarian hyperstimulation syndrome.

Ovarian hyperstimulation syndrome (OHSS) is a relatively common and potentially life-threatening complication of ovarian stimulation, the pathogenesis of which remains unclear. To clarify the predictive values of serum estradiol levels and oocyte number in severe OHSS, and to investigate the impact of high serum estradiol levels on pregnancy outcome, we retrospectively analyzed clinical data from 431 cycles of ovarian stimulation for assisted reproduction performed from 1993 through 1995. Receiver operating characteristic plots were used to estimate the predictive power of the measured variables. The overall frequency of severe OHSS was 5.5%. Using a serum estradiol level of 3,600 pg/mL as the minimum cut-off value, the sensitivity was 58%, with a specificity of 92%, a positive predictive value of 29%, and a negative predictive value of 97%. The predictive power was similar when a cut-off point of 20 oocytes retrieved was used. The two criteria together gave a sensitivity of 33%, a specificity of 92%, a positive predictive value of 40%, and a negative predictive value of 98%. One of seven oocyte donors developed severe OHSS. The pregnancy rate was higher in patients with severe OHSS than in patients who did not develop this syndrome (73.9% vs 32.5%) but the pregnancy outcomes were not significantly different. We conclude that elevated estradiol concentrations and oocyte number appear to be helpful in predicting severe OHSS, but neither parameter by itself is predictive. This syndrome is rare in the absence of luteal hCG support, either exogenous or pregnancy-derived; when it occurs, there are usually extremely high preovulatory estradiol concentrations and numerous oocytes retrieved. High serum estradiol levels are unlikely to have adverse effects on pregnancy outcome in patients with severe OHSS.

T lymphocytes and cytokine production in ascitic fluid of ovarian malignancies.

The activation status of T lymphocytes and the presence of various cytokines in ascitic fluid were examined to test peritoneal immunity in women with ovarian malignancies. Peripheral blood and peritoneal fluid were collected from 12 patients with primary ovarian cancer with ascites and 27 normal control subjects during laparoscopic examination. Lymphocyte subpopulations and the expression of activation markers on T lymphocytes were analyzed by dual-color flow cytometry. The concentrations of various cytokines and soluble interleukin (IL)-2 receptor-alpha were measured. CD8 T lymphocytes were the main component of peritoneal lymphocytes. CD69 and HLA-DR, but not CD25, were highly expressed on peritoneal T lymphocytes compared to those in peripheral blood. In ascitic fluid of ovarian malignancies, CD4 T lymphocyte concentrations were further decreased, resulting in a decreased CD4/CD8 ratio. Decreased expression of CD69 and CD25 was also noted on T lymphocytes from ascites compared with T lymphocytes in normal peritoneal fluid. IL-1b, tumor necrosis factor-alpha, IL-6, and soluble IL-2 receptor-alpha concentrations were increased significantly in the ascitic fluid of women with ovarian cancer. The decrease in activation markers on T lymphocytes is suggestive of an immunosuppressive state, despite the presence of abundant stimulatory cytokines. The immunosuppression may be multifactorial, attributed, in part, to the increased concentrations of soluble IL-2 receptor-alpha and other inhibitors.