Apoptotic DC-SIGN+ cells in normal human decidua.

BACKGROUND: Normal pregnancy and spontaneous abortion in humans and mice are associated with immune responses. The decidua harbors dendritic cells identifiable in humans by their expression of DC-SIGN. Because dendritic cells are essential for immune response regulation, decidual DC-SIGN+ cells may play a role in normal or pathological pregnancy outcomes. Previous reports suggested that DC interact with NK cells in decidua, although the functional significance of this phenomenon remains unknown. OBJECTIVE: We studied the presence of conjugates of DC-SIGN+ cells with CD56+ NK cells in normal human decidua. METHODS: Conjugates of DC-SIGN+ cells with CD56+ NK cells were studied in leukocyte suspensions of normal human decidua (6-11 weeks) by flow cytometry and confocal microscopy. The presence of apoptotic cells was determined by the TUNEL assay, incubation with annexin V and confocal microscopy in decidual leukocyte suspensions and by the TUNEL assay in decidual sections. RESULTS: We observed conjugates of decidual DC-SIGN+ cells with CD56+ NK cells (40.2±26.1% of all the DC-SIGN+ cells by flow cytometry and 52.3±10.2% by confocal microscopy). We also found that a proportion of DC-SIGN+ cells were in apoptosis, since they were TUNEL+ (40.2±7.2% of all DC-SIGN+ cells in decidual sections) and annexin V+ (34.4±15.2% in leukocyte suspensions). And sorted DC-SIGN+ cells had multilobulated nuclei. CONCLUSIONS: The conjugates of decidual DC-SIGN+ cells with CD56+ NK cells strongly suggest that these latter cells induce apoptosis in DC-SIGN+ cells during normal pregnancy. We discuss this possibility in the context of maternal-fetal tolerance.

Reduced proportion of decidual DC-SIGN+ cells in human spontaneous abortion.

Recent studies showed that some functions of decidual dendritic cells appear to be essential for pregnancy. In humans, decidual dendritic cells are identifiable by their expression of DC-SIGN. We compared the subpopulations of human decidual DC-SIGN+ cells from first-trimester normal pregnancies and spontaneous abortions by flow cytometry. In normal decidua, DC-SIGN+ cells expressed antigens associated with immature myeloid dendritic cells. In samples from spontaneous abortions, we detected decidual DC-SIGN+ cells with an antigen phenotype equivalent to that of DC-SIGN+ cells from normal pregnancies, but at a significantly lower proportion (P < 0.01). Our results support the hypothesis that dendritic cells play a role in normal or pathological human pregnancy outcomes.

Development of chronic colitis is dependent on the cytokine MIF.

The cytokine macrophage-migration inhibitory factor (MIF) is secreted by a number of cell types upon induction by lipopolysaccharide (LPS). Because colitis is dependent on interplay between the mucosal immune system and intestinal bacteria, we investigated the role of MIF in experimental colitis. MIF-deficient mice failed to develop disease, but reconstitution of MIF-deficient mice with wild-type innate immune cells restored colitis. In addition, established colitis could be treated with anti-MIF immunoglobulins. Thus, murine colitis is dependent on continuous MIF production by the innate immune system. Because we found increased plasma MIF concentrations in patients with Crohn's disease, these data suggested that MIF is a new target for intervention in Crohn's disease.

Effect of Halococcus morrhuae and Halobacterium saccharovorum on the activation of human peripheral blood lymphocytes.

The immunomodulator properties of two species of halophilic Archaebacteria, Halobacterium saccharovorum and Halococcus rnorrhuae, were analysed by the study of lymphocyte activation. Two methods were used to detect activation in lymphocytes, namely incorporation of the radioactive nucleotide [3H]-thymidine, and CD25 expression. H. morrhuae had a stimulatory effect on human lymphocytes, but this action was observed only with the [3H]-thymidine uptake method, whereas H. saccharovorum produced no immunomodulator effect.

Antigen phenotype of cultured decidual stromal cells of human term decidua.

We previously reported that decidual stromal cells (DSC) from early human decidua express antigens associated with hematopoietic cells and develop different immune functions. Here we study the antigenic phenotype of DSC from term decidua and compare it with the phenotype reported for DSC from early decidua. Decidual stromal cells were isolated from human term deciduas and maintained in culture until highly purified DSC cultures were obtained. Most term DSC, like most early DSC, expressed CD10. Term DSC expressed antigens specific for follicular dendritic cells (FDC), such as DRC-1 (CD21L) and HJ2, together with CD21, CD23 and CD80, which are detected on FDC as well. Also like early DSC, term DSC were negative for CD3, CD14, CD15 and CD45. Although early DSC were reported to be HLA-DR-positive and CD86-positive, these antigens were not expressed by term DSC. These discrepant results suggest that two types of cells, or cells at different stages of differentiation (decidualization) were selected during culture of decidual cells from different periods of gestation. This possibility was further supported by the finding that term DSC expressed desmin and prolactin, two markers of decidualization, whereas these molecules have not previously been detected in early DSC.

CD10 expression in cultured human osteoblast-like cells.

Morphological features, bone nodule formation and alkaline phosphatase activity are currently used to identify osteoblasts. CD10 (cALLa antigen) is a glycoprotein with endopeptidase activity and it is present on the surface of many cell types. We have studied the expression of CD10 in osteoblast-like cells by immunocytochemistry and flow cytometry in order to identify other markers of the osteoblast lineage. We isolated osteoblast-like cells from specimens obtained in the course of oral surgery. Expression of the cALLa antigen (CD10) may also be an indicator of the osteoblast phenotype.

Phagocytosis by fresh and cultured human decidual stromal cells: opposite effects of interleukin-1 alpha and progesterone.

Flow cytometry and transmission electron microscopy have been employed to show that a proportion of fresh and cultured human decidual stromal cells phagocytose latex particles. Phagocytosis of Escherichia coli by cultured decidual stromal cells was, however, very low. Stimulation of cultured decidual stromal cells with interleukin-1 alpha enhanced phagocytosis of both latex particles and E. coli. In contrast, when decidual stromal cells were cultured with progesterone under decidualizing conditions, phagocytic activity was reduced. These results suggest the existence of an immune-endocrine circuit involving decidual stromal cells.

Lymphocytes of human term decidua decrease cell adhesion to a plastic substrate.

Although human decidual lymphocytes have been widely studied, their function and possible interaction with trophoblast are still unclear. Here we show that whereas human early (EDL) and term (TDL) decidual lymphocytes were unable to kill human trophoblast by necrosis (assessed by the 51Cr-release assay) or apoptosis (DNA fragment assay), TDL but not EDL decreased trophoblast adhesion to a plastic substrate as determined by a [3H]thymidine assay. This effect, however, was not selective for trophoblast, as TDL also decreased the adhesion to plastic of human decidual stromal cells and HeLa cells. Our results suggest that TDL may play a role in placental detachment during parturition by decreasing trophoblast or decidual stromal cell adhesion.

Immune phenotype and cytotoxic activity of lymphocytes from human term decidua against trophoblast.

Flow cytometric data were used to compare the phenotype of term decidual lymphocytes and peripheral blood lymphocytes. Unlike peripheral blood lymphocytes, a significant percentage of CD3+, CD4+, CD8+ and CD16+ term decidual lymphocyte populations expressed the CD69 activation marker. The relative proportions of CD38 in CD3+, CD4+ and CD8+ populations were more than twice as large in term decidual lymphocytes as in peripheral blood lymphocytes. As reported for early decidual lymphocytes, the expression of CD38 and CD69 by term decidual lymphocytes suggests that these cells are also regionally activated. However, term decidual lymphocytes showed no spontaneous cytotoxicity against normal trophoblast or its tumoral counterpart, JEG cells. After stimulation with interleukin-2, these lymphocytes became cytotoxic, as did peripheral blood lymphocytes. The relevance of this latter result to the immune control of the physiological and pathological invasion of the decidua by the trophoblast is discussed.