Kinetics of murine decidual dendritic cells

Dendritic cells (DCs) are professional antigen presenting cells (APC) capable of induction of primary immune responses as well as immunologic tolerance. Myeloid and lymphoid subsets of murine DCs are able to shift cytokine responses of T cells toward Th2 and Th1 profiles respectively. Thus, DCs would be suitable candidates to mediate the balance of maternal immune responses to conception. We analyzed pregnancy-related variations in uterus and splenic DCs in a murine model. C57BL/6-mated Balb/c female mice with vaginal plugs were scarified at early, middle, and late pregnancy. Frozen sections of uterus and spleen at each stage of pregnancy were immunostained with CD11c- and MHC-II-specific antibodies. Two-color immunohistochemistry was also carried out using anti-CD11c and one of the antibodies against CD11b, CD8α, CD86, and DEC-205. Using morphometric analysis, the average density of DCs and relative percentage of myeloid (CD11c+, CD11b+) and lymphoid DCs (CD11c+, CD8α+) were determined at each stage. Our results showed that DCs are present throughout the pregnancy in decidua. The average density of decidual DCs at early pregnancy was significantly higher relative to middle and late gestation or to those of endometrial DCs of non-pregnant mice. Interestingly, the average density of decidual and splenic DCs, followed the same variations at different stages of pregnancy. The relative percentage of decidual lymphoid DCs (LDC) was significantly higher at mid-gestation when compared with other stages of pregnancy or non-pregnant mice. Inversely, the frequency of myeloid DCs (MDC) and the MDC/LDC ratio were statistically lower at the middle stage of pregnancy. A majority of decidual DCs expressed MHC-II and CD86. At early pregnancy, DCs were more concentrated subadjacent to the luminal epithelial layers, whereas at mid- or late gestation, DCs were randomly distributed in the stroma and around the epithelium. Mid-pregnancy period was a critical point with regard to splenic DCs kinetics, as both the average density of DCs and the frequency of MDCs decreased significantly when compared with early or late pregnancy, although the relative percentage of splenic LDCs did not change. Our data suggest that the balance of MDC and LDC is finely tuned throughout pregnancy, pointing an eminent immunoregulatory role of DCs in the maintenance of pregnacy. © 2007 Society for Reproduction and Fertility

Hydro-mechanical modelling of the boom clay excavation, convergence and contact with concrete lining

The Boom Clay is considered as one of the potential host rock formation in Belgium for radioactive waste repository in deep geological layers. Gallery excavations will induce large hydro-mechanical disturbances around disposal system that need to be well understood and characterised. This study discusses particularly the role of interactions between the lining of the galleries and the host formation in the numerical characterisation of excavations in Boom Clay. The excavation and the convergence of the connecting gallery of the HADES underground research facility in Mol is modelled in a hydro-mechanical framework. Zero-thickness interface elements are used to manage numerically the contact between the host rock and the lining. Numerical predictions are compared with strains measurements recorded within the concrete segments of the lining in the underground research laboratory in Mol. The study highlights the impact of the anisotropic behavior of the host rock on the response of the model.SCOPUS: cp.kinfo:eu-repo/semantics/publishe

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+ CD8α+) 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, CD8α, 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. © 2006 Elsevier Ireland Ltd. All rights reserved

The efficient isolation of murine splenic dendritic cells and their cytochemical features

Despite their importance in professional antigen presentation and their ubiquitous presence, dendritic cells (DCs) are usually found in such trace amounts in tissues that their isolation with high purity is a difficult task. Because of their scarcity, accurate determination of the purity of isolated dendritic cells is very important. In this study, we purified murine splenic dendritic cells by a three-step enrichment method and evaluated their morphological, cytochemical and functional characteristics. Purity of the isolated cells was determined by established methods such as flow cytometry (FC) and immunocytochemistry (ICC) using anti-CD11c monoclonal antibody. In order to test purified DC functional properties, we used in vivo antigen presentation assay. Our results showed that antigen-pulsed DCs are potent stimulators of antigen-specific lymphocyte proliferation. We studied myeloperoxidase (MPO) and non-specific esterase (NSE) activity in isolated cells to determine the purity of dendritic cells compared to more conventional methods. Our results showed that murine splenic dendritic cells were deficient in both MPO and NSE activity and the percentage of purity obtained by NSE staining on isolated cells was comparable to the results obtained by either FC or ICC. To our knowledge, this is the first report on using NSE activity for determination of the purity of isolated murine splenic dendritic cells. We, therefore, recommend that NSE activity be employed as a simple, inexpensive and yet accurate method for evaluation of the purity of isolated murine splenic dendritic cells. © Springer-Verlag 2006

Microenvironment of the feto-maternal interface protects the semiallogenic fetus through its immunomodulatory activity on dendritic cells

Objective: To investigate the immunomodulatory activity of decidual culture supernatant on dendritic cell (DC) functions. Design: In vivo and in vitro experimental study using mice. Setting: Academic research laboratory. Animal(s): C57BL/6-mated female Balb/c mice. Intervention(s): Culture supernatants of decidual cells obtained from the uteri of allogenic pregnant mice (Balb/c � C57BL/6) were collected. Dendritic cells were purified from Balb/c mice spleens and pulsed with antigen during overnight culture. In some cultures, decidual supernatant was added at 5, 10, or 20 final concentration. Endometrial culture supernatant-treated DCs served as a control. Antigen-pulsed DCs were injected into the front footpads of syngeneic mice. Main Outcome Measure(s): Lymph nodes of primed mice were removed 5 days after DC injection. Antigen-specific proliferation and interleukin-10 and interferon gamma production by lymphocytes were measured by 3H-Thymidine incorporation and ELISA, respectively. Result(s): The results showed that decidual culture supernatant markedly blocked in vivo antigen presentation by DCs and inhibited their capacity to induce interferon gamma (but not interleukin-10) production by primed lymphocytes. Conclusion(s): It seems that soluble factors produced by decidual cells are important mediators of immunoregulation at the feto-maternal interface, which provide the two fundamental requirements for protection of the semiallogenic fetus, namely immunologic tolerance and predominance of T helper 2 immunity, through modulation of DCs function. © 2008 American Society for Reproductive Medicine