Intradermal testing for autoimmune progesterone dermatitis: should we be basing the diagnosis on it?

Autoimmune progesterone dermatitis (APD) is a rare skin disorder with varying presentations, resulting from hypersensitivity to endogenous progesterone during the luteal phase of the menstrual cycle. The diagnosis has been traditionally confirmed with intradermal progesterone testing (IPT) or intramuscular challenge with progesterone or its derivatives. We present a case of a 31-year-old woman with suspected APD who underwent IPT to progesterone. The patient's cyclical symptoms, positive skin reaction and symptoms following IPT were sufficient to make a diagnosis of APD. However, we also tested 10 healthy female controls without symptoms of APD, and found that 9 of these also developed positive skin reactions to intradermal progesterone at 15 min, 24 and 48 h, albeit to a lesser extent. Therefore, these results raise doubts about the validity of using IPT to make a diagnosis of APD. Further research on appropriate testing is needed.

Serum IL-1RA levels increase from follicular to luteal phase of the ovarian cycle: A pilot study on human female immune responses.

The immune responses exhibited by females are distinct from those of males. Females are known to generate, among others, higher levels of antibodies, greater interferon responses, and increased levels of inflammatory mediators in response to pathogens. Mounting evidence suggests that gonadal hormones play a key role in these differences. To better understand the effect of cycling hormones on the immune response, we sought to investigate the relationship between gonadal hormone fluctuations during the ovarian cycle and the levels of interleukin 1ß and IL-1RA, both in circulation and in PBMCs in response to TLR4 stimulation, in healthy premenopausal females. To do this we measured the gonadal hormones 17ß-estradiol, progesterone, and luteinizing hormone, and the cytokines IL-1ß and IL-1RA in nine cycling females at several time points throughout one complete cycle. We evaluated 35 follicular, 17 ovulatory, and 44 luteal time points in our cohort and found a clear increase in serum levels of anti-inflammatory IL-1RA in the luteal phase, as compared to the follicular phase, and a positive correlation between both 17ß-estradiol and progesterone and IL-RA. There was no difference in the serum levels of IL-1ß and no difference in IL-1 ß or IL-1RA produced in response to LPS by PBMCs isolated from different phases. Division of the cycle into sub-phases revealed an increase in the level of IL-1RA by ovulation that persisted through the luteal phase. These data suggest that significant changes in the immune response occur throughout the ovarian cycle in healthy females.

Immunoglobulin J chain as a non-invasive indicator of pregnancy in the cheetah (Acinonyx jubatus).

The North American cheetah population serves as a reservoir for the species, and acts as a research population to help understand the unique biology of the species. Little is known about the intrauterine physiology of the cheetah, including embryo differentiation, implantation, and the development of the placenta. After mating, cheetah females frequently experience (30-65% of matings) a non-pregnant luteal phase where progestogen metabolite levels match those found in pregnant females for the first ~55 days of gestation, but parturition does not occur. Immunoglobulin J chain (IgJ) is a molecule that is involved in the activation of the secretory immune response and has been found to be indicative of pregnancy in the cheetah using fecal monitoring. In this study, western blotting was employed to track IgJ abundance in pooled weekly fecal samples following natural breeding or exogenous stimulation to ovulate, and IgJ levels were compared between individuals undergoing a pregnant (n = 12) and non-pregnant (n = 19) luteal phase. It was revealed that IgJ abundance was increased in pregnant females compared to non-pregnant females at week 4 and week 8 post-breeding, indicating the potential modulation of maternal immunity in response to sensitive events such as implantation and the increased secretory activity of the placenta. IgJ levels also tended to be higher early after breeding in females that were bred naturally with intact males compared to exogenously stimulated females with no exposure to seminal plasma, potentially indicating a response to the act of intromission or the stress of breeding, or possibly demonstrating an immune response resulting in the promotion of maternal tolerance to seminal antigens present upon embryonic implantation. Monitoring fecal IgJ may be a potential method to determine gestational status in the cheetah and will aid future conservation efforts of the species.

Endometrial immunocompetent cells in proliferative and secretory phase of normal menstrual cycle.

BACKGROUND: Menstruation was presented as a result of inflammatory process. The total and relative numbers of the endometrial immunocompetitive cells vary during the different phases of the menstrual cycle. The aim of this morphological study is to make a contribution to understanding different distribution of leukocyte types during proliferative and secretory phase of normal menstrual cycle. MATERIALS AND METHODS: The study included 40 women (20 in proliferative and 20 in secretory phase of the menstrual cycle). Exploratory curettage performed as preoperative preparation due to uterine myomas. Immunophenotyping was performed by immunoalkaline phosphatase (APAAP) using monoclonal antibodies: CD15, CD20, CD30, CD45RO, CD56, CD57 and CD68. The results were statistically analysed using SPSS 20.0 software. RESULTS: Natural killer (NK) cells are dominant during secretory, and CD45RO T lymphocytes are dominant during proliferative phase of the menstrual cycle. During the secretory phase of menstrual cycle, leukocytes make 30% of total endometrial cells. NK cells (CD56+ bright subpopulation), activated T lymphocytes, macrophages and B lymphocytes significantly increase in their number during the secretory phase of menstrual cycle. CONCLUSIONS: Significant changes in endometrial leukocyte populations during proliferative and secretory phase of the menstrual cycle are emphasized. Changes in dominance of different leukocyte subpopulations are determined by hormonal and microenvironmental changes in modulatory factors that have not yet been fully explained.

Changes in Functional Activity of Neutrophils and Monocytes Isolated from the Peripheral Blood of Women at Different Phases of the Menstrual Cycle.

We studied functional activity of neutrophilic granulocytes and monocytes isolated from the peripheral blood of women during the follicular and luteal phases of the menstrual cycle. It was shown that phagocytic activity of neutrophilic granulocytes increases, their intracellular oxygen-dependent bactericidal activity decreases, and the number of monocyte extracellular traps increases in women in the luteal phase of the menstrual cycle in comparison with the follicular phase.

Tissue distribution of some immune cells in bovine reproductive tract during follicular and luteal phase.

More recent studies indicate that immune cells which secrete their secretory products or cytokines play an important role in reproductive system. In our study, immune cell populations (CD8+ T lymphocytes, CD68+ macrophages, plasma cells, siderophages, eosinophils) and expression of major histocompatibility complex (MHC) class I and class II were examined in female reproductive tract during follicular (n = 13) and luteal phase (n = 10). Plasma cells and eosinophil granulocytes are present in few numbers in luminal epithelium, but abundant in longitudinal muscle layer of uterus, whereas siderophages are the dominant cell type in stroma. Moreover, MHC-I and -II+ cells are expressed by individual cells in organ layers, while CD8+ T cells and CD68+ macrophages are dominant in epithelium and muscle layer, respectively. In conclusion, we did not found significant changes in immune cells according to follicular and luteal phases, but localization and numbers in each organ have changed according to both organ and layers. These results indicate that these factors may play a crucial role not only to generate an immune response but also to have a role in regulation of physiological functions in female reproductive organs.

Expression of E2A in mid-secretory endometrium of women suffering from recurrent miscarriage.

E2A is involved in promoting forkhead box P3 (FOXP3) and retinoid-related orphan receptor gamma t (RORγt) gene transcription, which are pivotal transcription factors of T regulatory cells and Th17 cells, respectively. Little is known about the involvement of E2A in pregnancy process. This study aimed to investigate the expression of E2A, cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), and Foxp3 in luteal phase endometrium of women suffering recurrent miscarriage (RM) (n=21) and control group (n=11) by immunohistochemistry, with the Vectra® automated quantitative pathology imaging system for analysis. The percentage of E2A+ cells and CTLA-4+ cells was significantly higher in the endometrium of women with RM than in the controls. There was positive correlation between E2A and CTLA-4 (r=0.523, P=0.002), E2A and FOXP3 (r=0.380, P=0.032), and FOXP3 and CTLA-4 (r=0.625, P=0.000) in the mid-secretory phase of endometrium for all subjects. It was concluded that the abnormal expression of endometrial E2A existed in mid-secretory endometrium of women with RM, and there was a positive correlation between E2A and FOXP3, and E2A and CTLA-4, suggesting the possible regulation role of E2A involved in regulating endometrium receptivity.

Inflammatory and endothelial markers during the menstrual cycle.

BACKGROUND: The menstrual cycle exhibits a pattern of repeated inflammatory activity. The present study aims to evaluate inflammatory and endothelial markers during the two phases of a menstrual cycle. METHODS: The study cohort consisted of 102 women with regular menstrual cycles. Inflammatory and endothelial markers (interleukin-6 [IL-6], pentraxin-3 [PTX-3], hs-C reactive protein [hs-CRP], sE-selectin, sP-selectin, intracellular and vascular cell adhesion molecules [ICAM-1 and VCAM-1] and cathepsins L, B and S) were measured during the early follicular and the late luteal phase of a normal menstrual cycle. RESULTS: Pentraxin-3 (PTX-3) and hs-CRP were significantly higher during the follicular phase compared to the luteal phase (p < 0.001 respectively p = 0.025). The other inflammatory and endothelial markers, with the exception of cathepsin B, were higher, albeit not significantly, during the follicular phase. CONCLUSIONS: Inflammatory activity, expressed mainly by members of the pentraxin family, is higher during the early follicular compared to the luteal phase. This could be associated to menstruation but the exact mechanisms behind this pattern are unclear and might involve the ovarian hormones or an effect on hepatocytes.

Comparison of Follicular and Luteal Phase Mucosal Markers of HIV Susceptibility in Healthy Women.

The purpose of this study was to evaluate differences in vaginal immune cell populations, vaginal tissue gene expression, antimicrobial activity of the cervicovaginal (CV) lavage (CVL), vaginal flora, and p24 antigen production from CV tissues after ex vivo human immunodeficiency virus (HIV) infection between follicular (FOL) and luteal (LUT) phases of the menstrual cycle. CV tissue biopsies, CV secretions, and blood samples were obtained as part of two longitudinal clinical trials of healthy women (CONRAD D11-119 and A12-124 studies). Participants (n = 39) were HIV-seronegative women not using exogenous hormone supplementation, with normal menstrual cycles, who were screened to exclude sexually transmitted and reproductive tract infections. Serum levels of estradiol and progesterone were significantly higher in the LUT versus the FOL phase of the menstrual cycle. Controlling for race, reported contraceptive use/sexual practices, and clinical trial, we found no differences in vaginal tissue immune cell populations and activation status, transcriptomes, inhibition of HIV, herpes simplex virus type 2 and Escherichia coli by the CVL, vaginal pH or Nugent score, or production of p24 antigen after ex vivo infection by HIV-1BaL between CV samples obtained in the FOL phase versus the LUT phase of the menstrual cycle. There were no significant correlations between serum estradiol and progesterone levels and CV endpoints. The hypothesis that the LUT phase of the menstrual cycle represents a more vulnerable stage for mucosal infection with HIV was not supported by data from samples obtained from the lower genital tract (ectocervix and vagina) from these two clinical trials.

T regulatory markers expression in unexplained recurrent spontaneous abortion.

OBJECTIVE: To evaluate expression of glucocorticoid-induced tumor necrosis factor receptor (GITR), cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and IL-10 in peripheral blood mononuclear cells (PBMCs) of 20 women with unexplained recurrent spontaneous abortion (URSA) compared to 20 normal non-pregnant women (NNP) during luteal phase in the window of implantation. METHODS: Quantitative real-time PCR (qRT-PCR) was performed using the Taqman method for expression of GITR and SYBR Green method for expression of CTLA-4 and IL-10. RESULTS: Expression of CTLA-4 in the NNPs (median; interquartile range; 3; 1.8-10) was significantly higher than the URSAs (0.72; 0.26-3.81, p = 0.015). Expression of GITR in the NNPs (53; 10-139) was significantly higher than the URSAs (6; 3-27, p = 0.005). However, IL-10 expression in the URSAs was significantly higher than the NNPs, did not meet a significant value. A significant correlation was found between CTLA-4 and GITR expression in the study population (p = 0.0001). CONCLUSIONS: Expression of CTLA-4 and GITR were significantly down-regulated in the URSAs compared to NNPs at the window of implantation, which shows the essential role of Treg cells in creating an immunological privileged site for fetus as an allograft at the maternal-fetal interface by high expression levels of CTLA-4 and GITR during a normal pregnancy.

Dynamics of Immune Cell Types Within the Macaque Corpus Luteum During the Menstrual Cycle: Role of Progesterone.

The goal of the current study was to characterize the immune cell types within the primate corpus luteum (CL). Luteal tissue was collected from rhesus females at discrete intervals during the luteal phase of the natural menstrual cycle. Dispersed cells were incubated with fluorescently labeled antibodies specific for the immune cell surface proteins CD11b (neutrophils and monocytes/macrophages), CD14 (monocytes/macrophages), CD16 (natural killer [NK] cells), CD20 (B-lymphocytes), and CD3epsilon (T-lymphocytes) for analysis by flow cytometry. Numbers of CD11b-positive (CD11b(+)) and CD14(+) cells increased significantly 3 to 4 days after serum progesterone (P4) concentrations declined below 0.3 ng/ml. CD16(+) cells were the most abundant immune cell type in CL during the mid and mid-late luteal phases and were 3-fold increased 3 to 4 days after serum P4 decreased to baseline levels. CD3epsilon(+) cells tended to increase 3 to 4 days after P4 decline. To determine whether immune cells were upregulated by the loss of luteotropic (LH) support or through loss of LH-dependent steroid milieu, monkeys were assigned to 4 groups: control (no treatment), the GnRH antagonist Antide, Antide plus synthetic progestin (R5020), or Antide plus the estrogen receptor agonists diarylpropionitrile (DPN)/propyl-pyrazole-triol (PPT) during the mid-late luteal phase. Antide treatment increased the numbers of CD11b(+) and CD14(+) cells, whereas progestin, but not estrogen, replacement suppressed the numbers of CD11b(+), CD14(+), and CD16(+) cells. Neither Antide nor steroid replacement altered numbers of CD3epsilon(+) cells. These data suggest that increased numbers of innate immune cells in primate CL after P4 synthesis declines play a role in onset of structural regression of primate CL.

Molecular Signatures of Immune Activation and Epithelial Barrier Remodeling Are Enhanced during the Luteal Phase of the Menstrual Cycle: Implications for HIV Susceptibility.

UNLABELLED: The variable infectivity and transmissibility of HIV/SHIV has been recently associated with the menstrual cycle, with particular susceptibility observed during the luteal phase in nonhuman primate models and ex vivo human explant cultures, but the mechanism is poorly understood. Here, we performed an unbiased, mass spectrometry-based proteomic analysis to better understand the mucosal immunological processes underpinning this observed susceptibility to HIV infection. Cervicovaginal lavage samples (n = 19) were collected, characterized as follicular or luteal phase using days since last menstrual period, and analyzed by tandem mass spectrometry. Biological insights from these data were gained using a spectrum of computational methods, including hierarchical clustering, pathway analysis, gene set enrichment analysis, and partial least-squares discriminant analysis with LASSO feature selection. Of the 384 proteins identified, 43 were differentially abundant between phases (P < 0.05, ≥2-fold change). Cell-cell adhesion proteins and antiproteases were reduced, and leukocyte recruitment (interleukin-8 pathway, P = 1.41E-5) and extravasation proteins (P = 5.62E-4) were elevated during the luteal phase. LASSO/PLSDA identified a minimal profile of 18 proteins that best distinguished the luteal phase. This profile included cytoskeletal elements and proteases known to be involved in cellular movement. Gene set enrichment analysis associated CD4(+) T cell and neutrophil gene set signatures with the luteal phase (P < 0.05). Taken together, our findings indicate a strong association between proteins involved in tissue remodeling and leukocyte infiltration with the luteal phase, which may represent potential hormone-associated mechanisms of increased susceptibility to HIV. IMPORTANCE: Recent studies have discovered an enhanced susceptibility to HIV infection during the progesterone-dominant luteal phase of the menstrual cycle. However, the mechanism responsible for this enhanced susceptibility has not yet been determined. Understanding the source of this vulnerability will be important for designing efficacious HIV prevention technologies for women. Furthermore, these findings may also be extrapolated to better understand the impact of exogenous hormone application, such as the use of hormonal contraceptives, on HIV acquisition risk. Hormonal contraceptives are the most widely used contraceptive method in sub-Saharan Africa, the most HIV-burdened area of the world. For this reason, research conducted to better understand how hormones impact host immunity and susceptibility factors important for HIV infection is a global health priority.

Effect of the menstrual cycle on inflammatory cytokines in the periodontium.

BACKGROUND AND OBJECTIVE: The effects of different levels of steroid hormones, as experienced during puberty, pregnancy and menopause, on the periodontium have been demonstrated, but changes in sex hormone levels during the menstrual cycle, and the influence of these changes on the periodontium, remain unresolved. The aim of this study was to investigate the effect of the menstrual cycle on the levels of interleukin-1beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) in gingival crevicular fluid and on periodontal clinical parameters, including the gingival bleeding index (GBI) and the modified gingival index (MGI), in periodontally healthy women. MATERIAL AND METHODS: Twenty-seven periodontally healthy women with a regular menstrual cycle were included in the study. Clinical parameters, including the GBI, the MGI and the simplified oral health index, were recorded during menstruation, ovulation and premenstruation phases (e.g. on days 1-2, 12-14 and 22-24, respectively) of the menstrual cycle. Gingival crevicular fluid and unstimulated saliva were collected, at each study phase, for assessment of IL-1ß, TNF-α, estrogen and progesterone. RESULTS: Both the GBI and the MGI increased significantly during the menstrual cycle, and were significantly higher during ovulation than during menstruation or premenstruation (p < 0.001). No significant change in the simplified oral health index was observed during the menstrual cycle ( p = 0.18). The levels of IL-1ß and TNF-α increased during the different phases of the menstrual cycle, but only the change in the TNF-α concentration was significant ( p < 0.05). CONCLUSION: This study indicated that changes occurring during the menstrual cycle influence the periodontium and induce inflammatory conditions.

Dynamics of CD3⁺ T-cell distribution throughout the estrous cycle and gestation in the bovine endometrium.

T cells are the dominant lymphocytes in the endometrium and are considered to play a crucial role in implantation and in the maintenance of gestation through cytokine production and immune regulation. The mechanisms underlying immunoregulation at the feto-maternal interface are still obscure for this complex system. Understanding the role of T cells is a key factor in understanding the endometrial immune system. In this study, the distribution of endometrial CD3⁺ T cells in bovines was examined by immunohistochemical analysis. The estrous cycle and gestation was divided into 4 stages, and the number of CD3⁺-positive T cells was counted in each stage. CD3⁺ cells were found in the endometrium in significant numbers throughout the estrous cycle and were mostly located in the subepithelial area. The number of CD3⁺ cells significantly increased in the early and mid-luteal phases but decreased after implantation with the progression of gestation. No T cells were found in the placentome or specifically in the tissues near the fetus, including the trophoblastic area. In addition, very few T cells were found in stromal regions close to the myometrium of the endometrium. These findings suggest that downregulation of bovine endometrial CD3⁺ T-cell functions is closely related to the successful maintenance of gestation in a spatiotemporal manner.

The expression of B7-H1 and B7-H4 co-stimulatory molecules on myeloid and plasmacytoid dendritic cells in pre-eclampsia and normal pregnancy.

The aim of our study was to estimate the expression of B7-H1 and B7-H4 molecules on myeloid and plasmacytoid dendritic cells (DCs) in the peripheral blood of patients with pre-eclampsia, normal pregnant women and healthy non-pregnant women. Thirty-three patients with pre-eclampsia, 26 normal pregnant women, and 12 healthy non-pregnant women were included in the study. Dendritic cells were isolated from peripheral blood, stained with monoclonal antibodies against blood dendritic cell antigens and B7-H1 and B7-H4 molecules and estimated using flow cytometry. The expression of B7-H1 and B7-H4 molecules was significantly higher on CD1c(+) myeloid and CD303(+) plasmacytoid DCs in the first trimester of pregnancy than in the luteal phase of the ovarian cycle (CD1c(+)B7-H1(+): 19.19±10.55% vs. 11.99±6.79%; p<0.05; CD1c(+)B7-H4(+): 12.01±9.15% vs. 3.98±1.97%, p<0.001; CD303(+)B7-H1(+): 4.15±2.38% vs. 1.70±0.87%, p<0.05; CD303(+)B7-H4(+): 5.44±2.93% vs. 2.33±1.54%, p<0.01). Moreover, the expression of the B7-H1 molecule on CD1c(+) DCs in the second trimester of normal pregnancy was significantly higher than in the first trimester, but in the third trimester they decreased compared with the second trimester (II vs. I trimester: 32.23±11.30% vs. 19.19±10.55%, p<0.01; III vs. II trimester: 32.23±11.30% vs. 22.39±8.19%, p<0.01). The expression of B7-H1 molecule on CD1c(+) myeloid and CD303(+) plasmacytoid DCs was significantly lower in pre-eclampsia than in healthy third-trimester pregnant women (CD1c(+)B7-H1(+): 13.78±6.26% vs. 22.39±8.19%, p<0.05; CD303(+)B7-H1(+): 3.66±2.46% vs. 8.65±3.15%, p<0.01). Higher expressions of B7-H1 and B7-H4 molecules on CD1c(+) myeloid and CD303(+) plasmacytoid DCs in the first trimester of pregnancy suggest the role they play in the immunomodulation during early pregnancy.

Expansion of decidual CD45RO⁺ T cells with high expression of CEACAM1 in the early stage of pregnancy.

The aim of the present study was to investigate the mechanism involved in the expansion of CD45RO+ T cells in the decidual microenvironment, and in the expression of the inhibitory carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) on the surface of decidual CD45RO+ T cells. Twenty-one healthy nonpregnant females and seventeen healthy pregnant females in the first trimester were included in the study. Peripheral blood samples from nonpregnant and pregnant females, and decidual tissues from pregnant females following elective abortion, were obtained and analyzed by flow cytometry. The percentages of CD45RO+ T cells and CEACAM1-expressing CD45RO+ T cells were significantly higher in first trimester human decidua than in the peripheral blood. Conditioned medium from the coculture of monocytes and the human trophoblast HTR8/SVneo cell line (MHM) was added to the model for the generation of CD45RO+ T cells in vitro. MHM caused an increase in the percentage of CD45RO+ T cells in a monocyte chemoattractant protein-1 (MCP-1)­dependent manner and an increase in the percentage of CEACAM1-expressing CD4+CD45RO+ T cells in the model. In conclusion, our results implied that trophoblast cells and monocytes may be involved in the increase of decidual CD45RO+ T cells and the high expression of CEACAM1 on their surfaces.

Leucocyte phagocytosis during the luteal phase in bitches.

Pyometra is a disease that affects a large proportion of intact bitches, and typically is seen during the latter half of dioestrus. Several factors contribute to the development of pyometra, including genetic factors, an infectious component (most often Escherichia coli), and hormonal factors. Hormones may act directly on the endometrium, and also affect the immune system. In dogs, the phagocytic ability has been shown to decrease with age, and ovarian hormones have also been shown to affect immune resistance. The aim of the present study was to examine whether phagocytosis by canine leucocytes varies significantly during the luteal phase. Eight bitches were followed by repeated blood sampling. Samples were taken at the calculated optimal day for mating (Day 1), and thereafter on days 8, 15 and 22 (early luteal phase) and 29, 43, 57 and 71 (late luteal phase). Blood was collected from the cephalic vein into EDTA tubes for leucocyte counts and heparinised tubes for testing of phagocytosis and oxidative burst using commercial kits and flow cytometry. The cell activity of the phagocyting leucocytes, expressed as mean fluorescence activity, MFI, was significantly lower during late luteal phase than during early luteal phase. The proportion of leucocytes that was induced to phagocyte did not differ significantly. The percentage of cells stimulated by E. coli to oxidative burst was significantly lower during late luteal phase. Their activity did not differ between the two periods. The number of cells stimulated to oxidative burst by a low stimulus was too low to evaluate, and leucocytes stimulated with the high stimulus did not vary in oxidative burst between the two periods. The changes in phagocytic activity and in the number of leucocytes that showed oxidative burst were not associated with any change in the proportion of different leucocytes. The decreased phagocytic capacity possibly contributes to the higher incidence of diseases such as pyometra during the latter part of the luteal phase.

Decreased endometrial vascularity in patients with antiphospholipid antibodies-associated recurrent miscarriage during midluteal phase.

OBJECTIVE: To explore uterine arterial impedance and endometrial-subendometrial vascularity determined by two-dimensional Doppler ultrasonography (2D-DU) and three-dimensional ultrasonography and power Doppler angiography (3D-PDA) between patients with antiphospholipid antibodies-associated recurrent miscarriage (aPL-RM) and normal fertile women, and to further investigate the relationship between these parameters and endometrial microvessel density (MVD). DESIGN: Prospective observational study. SETTING: Tertiary-care fertility center. PATIENT(S): A total of 109 aPL-RM patients (aPL-RM group) and 49 normal fertile women (control group). INTERVENTION(S): Uterine measurement by transvaginal ultrasonography and endometrial MVD in the midluteal phase was assessed for both groups. MAIN OUTCOME MEASURE(S): Endometrial thickness, volume, and MVD, uterine arterial pulsatility index (PI) and resistance index (RI), and the vascularization index (VI), flow index (FI), and vascularization flow index (VFI) of endometrial and subendometrial regions were measured on day 7 of a natural cycle after ovulation. RESULT(S): Both groups had similar endometrial thickness, volume, and MVD and uterine arterial PI and RI. Endometrial VI, FI, VFI, and subendometrial FI were significantly reduced in the aPL-RM group compared with the control group. None of the uterine arterial 2D-DU and endometrial-subendometrial 3D-PDA parameters correlated with the endometrial MVD in both groups. CONCLUSION(S): Endometrial and subendometrial vascularity was significantly impaired in aPL-RM patients during natural midluteal phase compared with normal fertile women, and endometrial MVD did not correlate with any of the acquired vascularity parameters.

Toll-like receptor 4 expression in decidual cells and interstitial trophoblasts across human pregnancy.

PROBLEM: Toll-like receptor-4 (TLR-4) protects against Gram-negative bacteria expressed lipopolysaccharide and 'danger signals' from injured or dying cells. Although decidual cells (DCs) and interstitial trophoblasts (ITs) are in close contact, TLR-4 has been studied extensively only in ITs. METHOD OF STUDY: Formalin-fixed, paraffin-embedded serial sections of endometrium in follicular and luteal phases and deciduas from first and second trimester elective terminations and third trimester normal deliveries were immunostained for TLR-4, trophoblast-specific cytokeratin, and DC-specific vimentin. HSCORE assessed TLR-4 immunostaining in DCs versus ITs. RESULTS: TLR-4 HSCORES were significantly higher in: (i) first trimester DCs than luteal phase pre-decidual stromal cells; (ii) first and third versus second trimester DCs, but similar between third trimester deciduas parietalis and basalis; (iii) first versus second trimester ITs; (iv) DCs versus ITs across gestation. CONCLUSION: Higher TLR-4 in DCs than ITs suggests DCs as primary targets for Gram-negative bacteria and/or inflammation-related danger signals.

Menstrual cycle influences Toll-like receptor responses.

BACKGROUND: Toll-like receptors (TLRs) are pattern recognition receptors that play an important role as mediators of innate immunity. Human studies have shown changes in endometrial TLR expression during the menstrual cycle and during pregnancy. Our objective was to measure peripheral TLR activity over the course of the menstrual cycle. METHODS: We recruited 11 healthy females, and using ELISA we measured sex hormone levels and IL-1ß, IL-6, IL-8 and TNF-α following stimulation of whole blood with different TLR agonists during follicular, and early and late luteal phases of the menstrual cycle. RESULTS: During the follicular phase, we observed lower levels of IL-6 and TNF-α following stimulation with the TLR2 agonist HKLM when compared with the early luteal phase; lower levels of IL-1ß, IL-6 and TNF-α following stimulation with the TLR4 agonist LPS, and lower levels of IL-1ß and TNF-α following stimulation with the TLR5 agonist flagellin. Decreased IL-6 levels in the late compared to the early luteal phase were also observed following stimulation with the TLR5 agonist flagellin. Compared with the follicular phase, the late luteal phase of the cycle resulted in decreased levels of IL-1ß and TNF-α following stimulation with the TLR1/TLR2 agonist Pam3CSK and the TLR6/TLR2 agonist FSL1, as well as decreased levels of TNF-α following stimulation with the TLR8 agonist ssRNA40. There were no differences in cytokine release across the menstrual cycle following stimulation with the TLR3 agonist polyriboinosinic polyribocytidylic acid, or the TLR7 agonist Imiquimod. CONCLUSION: This study is the first to demonstrate that TLR responsivity in peripheral blood fluctuates throughout the menstrual cycle.