Immunosuppressive effect of progesterone on dendritic cells in mice

Progesterone has been demonstrated to be involved in maintaining pregnancy by regulating immunocytes. Dendritic cells (DCs), the most potent triggers of the adaptive immune response, express receptors for steroid hormones and are regarded as one of the primary targets of progesterone. However, the functional modification of DCs by progesterone remains poorly understood. Here, we report that progesterone does not affect the morphology or apoptosis of murine bone marrow-derived DCs. Progesterone-treated DCs were characterized by decreased expression of Ia (MHC class II), CD80 and CD86, increased production of IL-10, and decreased secretion of IL-12. Compared with mature DCs (mDCs), activated progesterone-treated DCs had a reduced capacity to stimulate CD4(+) T cell proliferation. The observation that progesterone-treated DCs could attenuate delayed-type hypersensitivity (DTH) responses in vivo suggests that progesterone mediates suppressive DC activity. However, transfer of progesterone-treated DCs into the peritoneal cavity of mice did not elevate the percentage of CD4(+)CD25(+)Foxp3(+) regulatory T cells in the spleen. Overall, our study helps to increase understanding of the role of DCs exposed to progesterone in the maintenance of pregnancy.     

Immunosuppressive activity of rhesus monkey pregnancy serum

Normal pregnancy serum from the rhesus monkey was found to have immunosuppressive activity. Using two-way stimulation, the mixed lymphocyte response was suppressed as much as 80%. Control serum from nonpregnant females was not suppressive. The inhibiting factor was found to have the following characteristics: (1) it was nonspecific in activity; (2) it inhibited the mixed lymphocyte response 20% at in vitro concentrations of 1%; (3) it was heat stable (56 degrees C for 30 minutes) and nondialyzable; (4) it was present in both the IgM- and IgG-containing fractions of pregnancy serum; (5) it was detected in postpartum and second and third trimester serum; and (6) it was at low levels or absent from the serum of two pregnancies which terminated in unexplained stillbirths.     

The effect of pregnant mare serum gonadotropin on mouse embryos fertilized in vivo or in vitro

The effect of increasing doses of exogenous gonadotropin stimulation for ovarian hyperstimulation was studied utilizing mouse embryos fertilized in vivo or in vitro. Increased rates of embryo degeneration, fragmentation, and triploidy, increased sister-chomatid exchange, and decreased fertilization rates were observed in high-dose stimulation groups. It appears, therefore, that oocyte and/or embryo quality may be affected by increased amounts of exogeneous gonadotropin stimulation.Presented in part at the American Fertility Society, New Orleans, Louisiana, April 2–7, 1984.     

Immunosuppressive effect of serum in patients with ovarian carcinoma.

Cell-mediated immune response was measured in 23 patients with ovarian cystadenocarcinoma, 38 patients with benign ovarian tumor, and 44 healthy volunteers. The method used two indexes: the lymphocyte response per unit volume of peripheral blood to phytohemagglutinin (PHA) and the immunosuppressive effect of serum on the response of normal lymphocytes to PHA stimulation. The lymphocyte response per 50 microliter peripheral blood did not differ significantly between patients with ovarian cancer and healthy volunteers. The serum effect, in contrast, differed significantly between malignant and benign ovarian tumors, and was found to increase significantly even when the cancer masses were as small as about 5 x 5 x 5 cm in size, ie, in FIGO Stage I. It is our belief that the measurement of the serum effect in patients with any ovarian tumor enables the early detection of ovarian cancer.     

早孕妇女血清及蜕膜组织液对自然杀伤细胞活性及淋巴细胞增殖的影响

目的　探讨早孕妇女血清及蜕膜组织液对自然杀伤 (NK)细胞及淋巴细胞功能的影响。方法　采用同位素掺入法 ,检测 3 2例正常早孕妇女血清 (早孕血清组 )及蜕膜组织液 (蜕膜组织液组 )对正常妇女外周血中NK细胞活性 (% )及淋巴细胞增殖功能的影响 ,并采用 2 5例正常未孕妇女的血清作对照 (对照组 )。结果　早孕血清组、蜕膜组织液组及对照组的NK细胞活性分别为 (3 5 .8±3 .0 ) %、(19.1± 2 .9) %及 (46.1± 3 .2 ) % ,淋巴细胞增殖功能 [每分钟同位素放射次数 (CPM) ]分别为(3 689± 4 5 6)CPM、(2 0 5 9± 2 88)CPM及 (4883± 64 3 )CPM ,3者间差异均有极显著性 (P <0 .0 1) ,且蜕膜组织液组均显著低于早孕血清组 (P <0 .0 0 5 )。结论　正常早孕妇女血清及蜕膜组织液具有免疫抑制作用 ,而蜕膜组织液可能是调节早期妊娠子宫局部免疫功能的重要因素之一。     

Immunosuppressive effect of serum progesterone during pregnancy depends on the progesterone binding capacity of the lymphocytes

Cytotoxic activity and progesterone binding capacity of the lymphocytes, together with serum progesterone concentrations, were determined in women with normal pregnancy or with a clinical diagnosis of threatened abortion or threatened premature labour. The lymphocytes of women with threatened abortion or threatened premature labour showed significantly higher cytotoxic activity (P < 0.001) and significantly lower progesterone binding capacity (P < 0.001) than did lymphocytes obtained from the healthy pregnant women. Significant inverse correlation was found between progesterone binding capacity and cytotoxic activity of the lymphocytes (P < 0.001), but the progesterone concentration of the pregnancy serum appeared to have no influence on the other two parameters. The findings indicate that intact progesterone binding capacity of the lymphocytes is an essential factor for the manifestation of the blocking effect exerted by pregnancy serum on lymphocyte cytotoxicity in vitro.     

Immunosuppressive properties of pregnancy serum on the mixed lymphocyte reaction

The human fetus may escape immunological attack because of serum factors which have immunomodulatory influence on maternal cellular effector responses. Paired peripheral and retroplacental sera were shown to inhibit the allogenic mixed lymphocyte reaction (MLR) used as an in-vitro model of cell-mediated immunity. There was no correlation between the suppressive effect of the peripheral and retroplacental sera and the serum concentrations of four pregnancy-related proteins (alpha-fetoprotein, pregnancy-associated alpha 2 glycoprotein, pregnancy-associated plasma protein A and Schwangerschaftsprotein 1) to which immunosuppressive properties have been ascribed, but there was a negative correlation between peripheral AFP and MLR inhibition (r = -0.62, P less than 0.001). Hence, the factor or factors responsible for suppressing the MLR are not those investigated in the present study.     

Immunosuppressive properties of pregnancy serum on the mixed lymphocyte reaction

Summary. The human fetus may escape immunological attack because of serum factors which have immunomodulatory influence on maternal cellular effector responses. Paired peripheral and retroplacental sera were shown to inhibit the allogenic mixed lymphocyte reaction (MLR) used as an in-vitro model of cell-mediated immunity. There was no correlation between the suppressive effect of the peripheral and retroplacental sera and the serum concentrations of four pregnancy-related proteins (α-fetoprotein, pregnancy-associated α₂ glycoprotein, pregnancy-associated plasma protein A and Schwangerschaftsprotein 1) to which immunosuppressive properties have been ascribed, but there was a negative correlation between peripheral AFP and MLR inhibition (r =−0·62, P < 0·001). Hence, the factor or factors responsible for suppressing the MLR are not those investigated in the present study.     

The effect of serum fractions on single-cell mouse embryos in vitro

To examine the effect of various fractions of human fetal cord serum (HCS) on mouse embryos cultured in vitro, heat-inactivated HCS was separated by ultrafiltration into five distinct fractions: Fractions A, MW>30,000; B, MW 30,000–10,000; C, MW 10,000–5000; D, MW 5000–1000; and E, MW <1000. Seven hundred twentyeight single-cell embryos were cultured in TYH- 280 medium supplemented with 8 mg/ml bovine serum albumin (BSA) and a 20% concentration of Fraction A, B, C, D, or E, whole HCS, or BSA alone. Embryos cultured with Fraction A or E or whole HCS demonstrated a significantly reduced growth rate (P<0.01), while embryos cultured with Fraction D demonstrated a significantly increased growth rate (P<0.01). Additionally, 649 singlecell embryos were cultured in medium which was supplemented with 8 mg BSA/ml and a 0, 1,2, or 5% concentration of Fraction A or E. Fraction E displayed toxicity even at a 1% concentration (P< 0.07), while Fraction A demonstrated growth inhibition at a 5% concentration (P <0.05) but increased the hatching rate at a 1% concentration (P < 0.01). Finally, 635 single-cell embryos were cultured with four distinct fractions of HCS obtained from a Sephacryl S-200 column: Fractions I, MW 100,000; II, MW 70,000–100,000; III, MW 30,000–70,000; and IV, low molecular weight (<5000). Fraction I or III significantly reduced the embryo growth rate as seen with Fraction A (P<0.01) and Fraction II significantly increased only the hatching rate (P<0.01), while Fraction IV significantly increased the growth rate as seen with Fraction D. In conclusion, HCS contains embryo growth inhibitory properties in the high (>30,000) and low (<1000) molecular weight components, while growth promoting factors are found in the 1000–5000 MW fraction. It also seems that there are some factors in the 70,000–100,000 MW fraction which may promote the ability of the embryo to hatch.Presented in part at the American Fertility Society meeting, March 1985.     

The effect of betamethasone administration to pregnant women on maternal serum indicators of infection

OBJECTIVE: To study the effect of betamethasone therapy on maternal white blood cell count, C-reactive protein and erythrocyte sedimentation rate in women at high risk for preterm delivery. STUDY DESIGN: We included women at gestational age of 24 to 34 weeks who were treated by betamethasone for enhancement of fetal lung maturity, because of imminent preterm labor with intact membranes. Blood tests for white blood cell and differential count, C-reactive protein and erythrocyte sedimentation rate were drawn before betamethasone injection, 2 hours after, and then every 24 hours for three days. RESULTS: 105 women were included. The mean white blood cell count increased by 33% on day one, and returned to baseline level three days after the first injection of betamethasone. A significant rise in neutrophil count, and drop in lymphocyte count was noted as early as two hours after the first injection and lasted for two days. Mean C-reactive protein and erythrocyte sedimentation rate levels were not changed significantly by betamethasone treatment. CONCLUSIONS: Antenatal betamethasone therapy causes a transient increase in maternal leukocyte count but has no effect on serum C-reactive protein and erythrocyte sedimentation rate. This information is relevant for preterm pregnant women who are at high risk for chorioamnionitis.     

C1q-binding activity of serum from allogeneically and syngeneically pregnant mice

C1q-binding activity of serum during allogeneic and syngeneic pregnancy in mice was measured using the direct [125I]C1q-binding assay. In both strains there was an initial tendency for levels to rise in very early pregnancy (Days 2 and 4) followed by a progressive fall in C1q binding to a level significantly lower than that found in non-pregnant animals. The changes were present in both syngeneic and allogeneic pregnancies but the later changes were more pronounced in the latter. The presence of changes associated with syngeneic pregnancy suggests that these profiles are at least in part independent of genetic differences between mother and embryo.     

Immunosuppressive activity of proteases in cervical carcinoma

OBJECTIVE: The host immune response is essential for restraining both HPV infections and HPV-related cervical cancer. We previously reported a direct correlation between proteolytic activity and malignant progression from precursor lesions to invasive cervical carcinoma. The present study was undertaken to investigate whether proteinases from cervical carcinoma extracts and representative purified proteinases involved in tumor progression could regulate lymphocyte proliferation to phytohemagglutinin (PHA) mitogen. METHODS: Extracts were prepared from tissue samples obtained from patients with invasive cervical squamous carcinoma, squamous intra-epithelial lesions or women with normal cervix. Lymphocytes obtained from a single healthy donor were pre-incubated with one of these extracts in the presence or absence of proteinase inhibitors, and stimulated with PHA during 72 h. The proliferative response was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) method (re-validated with thymidine uptake). RESULTS: Lymphocyte proliferation was significantly decreased by cervical carcinoma extracts, while only slightly decreased by squamous intra-epithelial lesions or normal extracts. Inhibitor assays indicated that proteinases from cervical carcinoma were responsible for 53.30% of total suppressive activity. We found that purified enzymes such as trypsin, cathepsin B, uPA and type IV collagenase suppressed the proliferative response in a dose-dependent fashion. CONCLUSIONS: Our data suggest that in addition to the classic role in tumor invasion, proteases could represent an immune evasion mechanism in cervical carcinoma.     

Analysis of endometrial myeloid and lymphoid dendritic cells during mouse estrous cycle

This study was performed to evaluate the frequency and localization of endometrial myeloid (CD11c(+) CD11b(+)) and lymphoid (CD11c(+) CD8alpha(+)) dendritic cells (DCs) at different stages of murine estrous cycle. To address the systemic effect of ovarian hormones fluctuations during estrous cycle, the same variables were studied in splenic DCs as well. Stages of the estrous cycle of Balb/c mice were determined by examination of vaginal smears. Frozen sections of uterus and spleen at each stage of estrous cycle were stained for CD11c and MHC-II. Two-color immunohistochemistry was also carried out using anti-CD11c with one of the antibodies against CD11b, CD8alpha, CD86, and DEC-205. The average density of DCs and relative percentage of myeloid and lymphoid DCs (MDCs and LDCs) were determined at each stage of estrous cycle by morphometric analysis. Our results showed that DCs were present throughout the estrous cycle in mice endometrium, but their frequency was highest at estrus and lowest at proestrus (P<0.005). The lymphoid subset of DCs was more prominent at estrus relative to those at other stages (P<0.005). Conversely, the relative percentage of myeloid DCs at estrus was significantly lower compared to other stages (P<0.005). Nearly all endometrial and splenic DCs expressed CD86 and MHC-II. At proestrus, and particularly at estrus, DCs were more concentrated subadjacent to the luminal and glandular epithelial layers with some scattered throughout the stroma whereas, at metestrus and diestrus, DCs were randomly distributed in stroma and around the glandular and luminal epithelial layers. The number and immunophenotype of splenic DCs were not statistically different between stages of estrous cycle. Our results suggest that endometrial but not splenic myeloid and lymphoid DCs are influenced by steroid hormones during estrous cycle.     

Investigation of effects of pregnant mare serum gonadotropin (PMSG) on the chromosomal complement of CD-1 mouse embryos

Purpose: The objective of this study was to examine the effect of superovulatory doses of gonadotropins on the frequency of chromosomal abnormalities of mouse embryos. Methods: Chromosome analysis of 8- to 16-cell stage mouse embryos and zygotes was performed by a cytogenetic method. Results: There was no significant effect of the pregnant mare serum gonadotropin (PMSG) dose on the level of aneuploidy and structural abnormalities from 8- to 16-cell-stage embryos among superovulated groups. However, a simple dose-response relationship between the PMSG dose and the incidence of polyploidy was observed, with the level of polyploidy rising from 2.9% with 10 IU PMSG to 10.5% with 15 IU PMSG. In zygote stage, the proportion of polyploid embryos also increased as the dose increased, from 1.9% in 5 IU to 6.7% in 15 IU PMSG. It was observed that the extra chromosomal set in polyploidy embryos originated by both fertilization of a diploid oocyte and dispermy. Conclusions: These results indicate a dose-response relationship between the PMSG dose and the incidence of polyploidy in the CD-1 mouse. Both a disturbance at maturation division and an error at fertilization were the cause of polyploidy.     

Immunosuppressive 30-kDa protein in urine of pregnant women and patients with trophoblastic diseases

Urine samples obtained from normal pregnant women and patients with trophoblastic diseases contain 30-kDa protein that suppresses phytohemagglutinin-induced T cell proliferation. The immunosuppressive protein was measured by a newly developed radioimmunoassay. The 30-kDa protein was demonstrated in almost all urine samples examined, fluid from hydatid vesicles and chorionic extracts, but not in any serum samples except at low levels in some sera from patients with choriocarcinoma. During pregnancy, the level of urinary 30-kDa protein was higher in the first (1625.5 ± 1212.0 ng/ml, mean ± S.D.) and second (1457.4 ± 1332.4 ng/ml) trimesters than in the third trimester (460.6 ± 419.0 ng/ml). The urinary 30-kDa protein/hCG ratios in patients with choriocarcinoma (8.3 ± 10.9) were significantly higher than those in patients with hydatidiform mole (0.67 ± 1.00, P < 0.01) and in all trimesters than those of normal pregnant women (0.54 ± 0.44 in the first trimester, P < 0.05; 0.63 ± 0.46 in the second trimester, P < 0.05; 0.24 ± 0.17 in the third trimester, P < 0.01). There is no significant difference between the ratios in hydatidiform mole and normal pregnancy. These findings and the fast disappearance of the 30-kDa protein from the circulation suggest that the 30-kDa protein plays a part in proliferation of trophoblastic cells in, or their invasion into the host by locally suppressing the immune reaction of the host and that the increase in the urinary 30-kDa protein level, in cases of choriocarcinoma, may be due to the malignant transformation of trophoblastic cells resulting in their rapid invasion.