Concepción Ruiz

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.

Antigenic Phenotype of Cultured Human Osteoblast-Like Cells

Background/Aims: Osteoblasts are classically considered to play an important role during bone tissue development, and to be involved in the formation of mineralized bone matrix. Recent reports have suggested that they can also exert some activities directly associated with the immune system (cytokine synthesis and antigen presentation). Moreover, some authors have found antigens on osteoblast-like cells normally expressed by other cells with a common origin in bone marrow.Methods: We isolated and cultured human osteoblast-like lines and studied their antigenic phenotype with flow cytometry using monoclonal antibodies against antigens associated with hematopoietic cells.Results: Cultured cells expressed CD34, but were negative for CD45. B cell antigens CD20 and CD23 and myelomonocytic antigens CD11b, CD13, and CD16 were detected. Expression of CD3, CD14, CD15 and CD68 was negative, whereas CD25 expression was positive. CD56, an antigen expressed on NK cells, was positive. These cells were CD10, CD44, CD54, CD80, CD86 and HLA-DR positive, as previously described. An antigen specific to follicular dendritic cells was also observed on cultured osteoblast-like cells.Conclusions: The antigenic phenotypes of human osteoblast-like cells and FDC are similar. These data suggest that osteoblasts may be functionally related to certain dendritic cells and may play an additional role in bone tissue to that classically assigned.

Constitutive Secretion of Interleukin-6 by Human Decidual Stromal Cells in Culture. Regulatory Effect of Progesterone

PROBLEM: Although several studies have demonstrated that decidual stromal cells (DSC) can secrete cytokines in culture, none of these studies documented the purity of the cultures. Since other cells of the decidua, such as macrophages and epithelial cells, also produce cytokines, it is important to ensure purity of the culture so that cytokine production can be ascribed with confidence to DSC.

Expression of cytokines IL-4, IL-12, IL-15, IL-18, and IFNγ and modulation by different growth factors in cultured human osteoblast-like cells

The antigenic phenotype of cultured human osteoblast-like cells, their ability to phagocytose particles of different nature and size, and their capacity to stimulate allogeneic T cells suggest that they are related to other cell populations with which they may also have immunological functions in common. The objective of this study was to investigate the intracytoplasmatic presence of cytokines and their modulation by different biomolecules. Immunocytochemistry and flow cytometry were used to study the expression of IL-4, IL-12, IL-15, IL-18, and IFNγ cytokines. To investigate whether FGF, TGF, PDGF, IL-1, and IFNγ modulate expression of these cytokines in cultured human osteoblast-like cells we used flow cytometry. IL-4, IL-12, IL-15, IL-18, and IFNγ cytokines were expressed by all the cultured human osteoblast-like cells studied. Treatment with FGF and TGFβ1 reduced the percentage expression and fluorescence intensity of the cytokines. PDGF treatment enhanced their fluorescence intensity but did not modify their expression. IL-1 treatment produced a small reduction in expression and fluorescence intensity of IL-12 and IL-15, but did not produce major changes in the expression of IL-4, IL-18, or IFNγ. IFNγ markedly increased the fluorescence intensity of the cytokines. The results indicate that human osteoblast-like cells may perform immunological functions (e.g., synthesizing cytokines with immune regulator function) that can be modulated by different biomolecules related to bone tissue and/or immune response.

Comparison of the Proportions of Leukocytes in Early and Term Human Decidua

The percentages of cells expressing immune markers were determined with immunohistochemistry and flow cytometry in early and term human decidua. Although we found no variation in the proportion of cells of bone marrow origin (CD45 +), the percentages of T cells and CD 16 + lymphocytes were significantly higher in term decidua. On the contrary, CD56+ lymphocytes, the most abundant leukocyte type in early decidua, decreased at term. These variations may reflex the immunological adaptations of decidua during pregnancy.

Phagocytosis and Allogeneic T Cell Stimulation by Cultured Human Osteoblast-Like Cells

Background/Aims: The antigen phenotype of human osteoblast-like cells suggests that they are related to other cellular populations and may also have immunologic functions in common. Methods: Flow cytometry and transmission electron microscopy were used to show the phagocytotic activity of osteoblast-like cells in culture. The allogeneic stimulation of T cells by human osteoblast-like cells was determined by the measurement of T cell proliferation. Results: We demonstrated in vitro that human osteoblast-like cells isolated from normal bone specimens obtained during mandibular osteotomy can phagocytose particles of different nature and size and can stimulate allogeneic T cells. Phagocytosis of microorganisms (E.coli, Klebsiella or C. albicans) was observed, although at a very low rate of activity in comparison with the phagocytosis of latex particles. Conclusion: Our results suggest that human osteoblast-like cells may perform immunologic functions and act as antigen presentation cells.p>

Massage after exercise--responses of immunologic and endocrine markers: a randomized single-blind placebo-controlled study.

Arroyo-Morales, M, Olea, N, Ruíz, C, Luna del Castilo, JD, Martínez, M, Lorenzo, C, and Díaz-Rodríguez, L. Massage after exercise-Responses of immunologic and endocrine markers: a randomized single-blind placebo-controlled study. J Strength Cond Res 23(2): 638-644, 2009-The effectiveness of massage for postexercise recovery remains unclear, despite numerous studies on this issue. The aim of this study was to determine the effect of massage on endocrine and immune functions of healthy active volunteers after intense exercise. After repeated Wingate tests, the effects of whole-body massage and placebo on salivary cortisol, immunoglobulin A (IgA), and total protein levels were compared using a between-group design. Sixty healthy active subjects (23 women, 37 men) underwent 2 exercise protocol sessions at least 2 weeks apart and at the same time of day. The first session familiarized participants with the protocol. In the second session, after a baseline measurement, subjects performed a standardized warm-up followed by three 30-second Wingate tests. After active recovery, subjects were randomly allocated to massage (40-minute myofascial induction) or placebo (40-minute sham electrotherapy) group. Saliva samples were taken before and after the exercise protocols and after recovery. In both groups, the exercise protocol induced a significant increase in cortisol (p < 0.001), decrease in salivary IgA (sIgA) (p < 0.001), and increase in total proteins (p = 0.01) in saliva. Generalized estimating equations showed a significant effect of massage on sIgA rate (p = 0.05), a tendency toward significant effect on salivary total protein levels (p = 0.10), and no effect on salivary flow rate (p = 0.55) or salivary cortisol (p = 0.39). The sIgA secretion rate was higher after the recovery intervention than at baseline among women in the massage group (p = 0.03) but similar to baseline levels among women in the placebo group (p = 0.29). Massage may favor recovery from the transient immunosuppression state induced by exercise in healthy active women, of particular value between high-intensity training sessions or competitions on the same day.

Antigenic Phenotype and Phagocytic Capacity of MG-63 Osteosarcoma Line

Human osteoblasts isolated from bone tissue samples have their own specific antigen profile but also share expression of antigens that are characteristic of other immunocompetent cells. Given that these findings come from studies performed in primary cultures of human osteoblasts, it was decided to test whether these antigen profiles and functional characteristics are retained in a characterized osteoblast cell line (MG‐63). We show that some of these characteristics are also found in the MG‐63 osteosarcoma cell line. We have demonstrated, using monoclonal antibodies and cytometry, that these cells expressed CD10, CD13, CD44, and CD54 antigens but were negative for CD69 and HLA‐DR antigens. Functionally, 100% of MG‐63 cells showed phagocytic capacity with a high phagocytic index. This study corroborates that osteoblastic cells have an immunological profile.

Cultured human decidual stromal cells express antigens associated with hematopoietic cells

Although decidual stromal cells (DSC) have classically been considered to play a nutritional role during pregnancy, several reports have demonstrated that they can also exert different immune activities. Furthermore, some authors have occasionally found antigens on DSC normally expressed by immune cells. In this study, we isolated and cultured 12 human DSC lines and studied them with immunocytochemistry and flow cytometry using monoclonal antibodies against antigens associated with hematopoietic cells. Decidual stromal cells exhibited a constant phenotype: they were CALLA (CD10)-positive and DR-positive, although the expression of CD45, the leukocyte common antigen, was found to be very weak or negative. We also detected myelomonocytic antigens CD11b (CR3), CD13, CD16 (Fc gamma RIII) and CD36, although DSC lacked CD14, CD15 and CD33. B cell antigens CD20, CD21 (CR3), CD23 (Fc epsilon RII) and CD24 were expressed. DRC-1, an antigen detected on follicular dendritic cells (FDC), was also observed on DSC. When these cells were cultured in the presence of progesterone, they expressed desmin and prolactin (PRL), findings that confirmed their identity as DSC. The phenotype described, together with the immune activities reportedly carried out by DSC, suggest that DSC may play a role in the maternal-fetal immune relationship.

Effects of Indomethacin, Nimesulide, and Diclofenac on Human MG-63 Osteosarcoma Cell Line

Nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most widely prescribed drugs worldwide and serve as treatment of some degenerative inflammatory joint diseases. The aim of the present study was to investigate the influence of different concentrations of three NSAIDs on cell proliferation, differentiation, antigenic profile, and cell cycle in the human MG-63 osteosarcoma cell line, incubated for 24 hr. All NSAIDs had an inhibiting effect on osteoblastic proliferation. Treatments for 24 hr had small but significant effects on the antigenic profile. No treatment altered osteocalcin synthesis. Indomethacin and nimesulide treatments arrested the cell cycle at G0/G1. These results suggest that indomethacin, nimesulide, and diclofenac appear to have no effects on osteocalcin synthesis and a slight effect on the antigenic profile. They may delay bone regeneration due to their inhibiting effect on osteoblast growth. Therefore, these drugs should only be used in situations that do not require rapid bone healing.