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Biomedical subjects

S Vassiliadis

Publications and source records attributed to S Vassiliadis.

At least 19 recordsLinked to original sources
PubMed2005

Constitutive or induced elevated levels of L-carnitine correlate with the cytokine and cellular profile of endometriosis.

During the past decade, accumulated evidence indicates an association between endometriosis and an alteration of humoral and cell-mediated immunity. While the role of L-carnitine in the regulation of energy metabolism is well established, it is only recently that L-carnitine has been recognized to modify the immune response in mice after in vitro or in vivo treatment. The present study has examined whether administration of L-carnitine to young female mice alters the percentage of immune cells in peritoneal exudates and the uterus as well as the levels of IFN-gamma, TNF-alpha, IL-2, IL-4, IL-6, VEGF, GM-CSF and IGF-I in blood serum, peritoneal fluid and supernatants of uterine cultured cells as tested by immunofluorescence or ELISA techniques, respectively, leading to a pathological disorder resembling human endometriosis. The results showed that, except from infertility, L-carnitine treatment resulted in a significant increase of macrophages and to a lesser degree an increase of T-cells, while elevated levels of IFN-gamma and TNF-alpha were detected in both serum and peritoneal fluid compared to controls. Although levels of L-carnitine measured in mouse serum samples using a radioisotopic method showed an increase as compared to controls, levels of acyl-L-carnitine measured in the murine peritoneal fluid samples showed a decrease similar to that measured in peritoneal fluid samples from patients with endometriosis in stage IV of the disease. These results indicate that L-carnitine administration to female mice alters the cellular and growth factor profile in the uterus and peritoneum towards a phenotypical pathology similar to that of clinical endometriosis.

Animals↗
PubMed2005

Endometriosis and infertility: a multi-cytokine imbalance versus ovulation, fertilization and early embryo development.

Endometriosis is tightly linked to infertility which is manifested at very early or more advanced stages of the gestational cycle. Alteration on the production of a great number of cytokines/growth factors can be accused for problems on ovum maturation, fertilization or implantation. Yet, macroscopically these stages are characterized by the inability of conception. A closer look of the cytokinic profile during the conceptional and early gestational cycle could, however, localize the problem and allow a therapeutic approach. In this commentary, going through the cytokine requirement during ovulation, fertilization and the early stages of pregnancy, it became possible to specifically define the harmful endometriosis-induced cytokines for each of the conceptional and early gestational stages. Thus, regulating the levels of interferon-gamma and tumor necrosis-alpha will facilitate ovulation and fertilization, whereas adjusting the levels of interleukin-1beta and colony stimulating gactor-1 will facilitate implantation.

Animals↗
PubMed2003

Serum concentrations of growth factors in women with and without endometriosis: the action of anti-endometriosis medicines.

Endometriosis is a common gynecologic syndrome of unknown etiology and pathogenesis. Growth factors and inflammatory mediators produced by peritoneal leukocytes have recently been postulated to participate in the pathogenesis of endometriosis. Angiogenic factors released from peritoneal macrophages may also play a role in the development of this disease. In the present study, we investigate the soluble levels of vascular endothelial growth factor (VEGF), epidermal growth factor-receptor (EGF-R), granulocyte/macrophage-colony stimulating factor (GM-CSF), Insulin-like growth factor-1 (IGF-1) and interferon-gamma (IFN-gamma) in the serum of 28 women with and 20 without endometriosis. We also compared these levels before, during and after treatment with danazol and leuprorelin acetate depot, the two therapeutic regiments of choice concerning this disease. We found that only sVEGF levels were higher in women with endometriosis in comparison to controls (P < 0.001) while sEGF-R is not present. GM-CSF, IGF-1 and IFN-gamma soluble levels are not affected in either healthy or endometriotic subjects. The 6-month treatment with danazol decreased sVEGF levels (P < 0.02) and increased sEGF-R levels (P < 0.001). These observations support the view that VEGF may be associated with the disease process and that danazol may bring sVEGF levels to a normal threshold. However, future studies will be focused on the anti-angiogenic control of the action of VEGF in patients with endometriosis.

Adult↗
PubMed2002

T-regulatory cells: are we re-discovering T suppressors?

Regulatory T cells are shown to originate form the thymus and their role is to maintain self-tolerance to intra-thymic as well as extra-thymic self-antigens. Their mode of action, using in vivo and in vitro systems, has led to different conclusions as to the need of cell-cell interactions or regulation upon suppressive cytokines. The more we study regulatory T cells the more we find similarities to the old notion of the suppressor T cell network. The limited knowledge in molecular technology in the early 70s and 80s discouraged investigators to further scrutinize the issue and the terms T suppressors and contra-suppressors that were coined back then have been forgotten over the years. It is now time to remember the work of these investigators and attempt to explain their findings using the current knowledge and technology.

Animals↗
PubMed2002

L-carnitine accelerates the in vitro regeneration of neural network from adult murine brain cells.

The development, growth and regeneration of nerve cells remain an unresolved issue. The up-to-date reported brain repair mechanisms are numerous and evidence suggests that, apart from the required trophism, tropism, microenvironment and specificity of the brain, a plethora of chemical, physiological and immunological compounds can contribute to such events. Among these compounds, we concentrated our interest on L-carnitine (L-Cn), which regulates the beta-oxidation of long chain fatty acids necessary for brain development, myelinization and growth. In contrast to fetal brain cells that grow easily in culture, adult brain cells show limited neurogenesis. Here, using adult brain cells from experimental mice, we show that although L-Cn does not improve their proliferative activity in short-term cultures, it accelerates the growth and differentiation of neurons, astrocytes, oligodendrocytes and ependymal cells from neurospheres in long-term cultures. Thus, the formation of a confluent neural network requires a 2-month period in culture. These observations provide new insights for in vivo use of L-Cn to support brain cell development in cases of injury or brain degenerative diseases.

Animals↗
PubMed2002

The role of L-carnitine on a restricted number of myeloid leukemia progenitor cells: generation of atypical cell types.

AIM: Since cellular maturation largely depends on lipid metabolism, we examined whether L-carnitine (L-C), a substance involved in these biochemical pathways, is able to promote differentiation of the promyelocytic cell line HL-60. METHODS: Differentiation was assessed by marker analysis, morphology, immunohistochemistry, proliferation and cellular activity assays. RESULTS: L-C increases HLA-DR and CD14 surface antigens, while morphologic and marker analysis of the treated cells reveals the presence of monocytes, neutrophiles and few dendritic cells. What is important, however, is the induction of cells that have an atypical to this pathway allure staining positive for the neurofilament 3A10 monoclonal antibody, specific for nerve cells and the anti-p75 (Nerve Growth Factor Receptor) monoclonal antibody. The events described concern active and, at the same time, not proliferative senescent cells. CONCLUSIONS: L-C exerts its differentiation action on a certain fraction of the leukemic population yielding a non-negligible number of atypical for the myeloid lineage cells. These findings complement earlier and recent reports that describe the generation of cells of a different lineage irrelevant to their parent line of differentiation indicating that the hemopoietic pool appears to be the source of any kind of cell types according to the stimulus provided. Thus, in the context of the plasticity theory it appears that the HL-60 cell line also possess the potential to differentiate towards unexpected pathways.

Carnitine↗
PubMed2001

Murine ectoplacental cone-derived trophoblast cells express chemokine receptors.

Chemokine receptors (CCRs) have been shown to regulate T cell migration and differentiation as well as the establishment of Th1/Th2 bias. Furthermore, T cells and T cell products are essential to trophoblast development. Thus, postulating that chemokines as well as their receptors may be expressed by trophoblast to move T cells into an interaction with the feto-placental unit, we examined whether CCRs are expressed during the early stages of ectoplacental cone (EPC) formation. For this, murine EPC-derived trophoblast were examined for their ability to express CCRs constitutively or inducible by interferon-gamma (IFN-gamma). Immunofluorescence experiments on EPC-derived trophoblast cells showed that CCR3, CXCR4 and CCR5 are significantly expressed. IFN-gamma accelerated the mobilization of intracellular pools of CCR molecules during early cell culture periods (2-6 h) and, in most cases, increased their expression on EPC-derived trophoblast cells. CCR activity could be detected in the culture supernatants of these cells, inversely proportional to cell surface expression, suggesting the existence of rapid endocytosis and recycling mechanisms. This finding indicates that the level of intracellular CCRs may partly be determined in the extracellular matrix, an event that could play an important role towards neutralization of specific T cell/trophoblast interactions during early stages of pregnancy and protect the fetus against harmful maternal immune responses.

Animals↗
PubMed2001

Soluble ICAM-1 levels in the serum of endometriotic patients appear to be independent of medical treatment.

Adhesion molecules regulate the interaction of cells with the extracellular matrix and/or other cells. The intercellular adhesion molecule-1 (ICAM-1; CD54) is a member of the immunoglobulin superfamily and expressed by several cell types, including leukocytes and endothelial cells. A circulating form of the usually membrane-bound molecule was identified and characterized in normal human serum and in sera from patients with endometriosis. In the present study, we established the serum-soluble ICAM-1 (sICAM-1) levels in patients with endometriosis. We also studied the effect of danazol and leuprorelin acetate depot on the levels of sICAM-1. Thirty-eight women, 18-45 years of age, with regular menses and documented pelvic endometriosis were recruited from a University Hospital setting. Twenty-two women with endometriosis were randomly divided into two groups. Danazol (600 mg) were given every day for 6 months, and 3.75 mg of leuprorelin acetate depot every 28 days for 6 months. Serum sICAM-1 concentrations were measured before, during and after treatment, and its quantitative determination was performed by an ELISA technique using a specific immunoassay. We found that (1) sICAM-1 levels were higher in women with endometriosis in comparison to healthy subjects; (2) the 6 month treatment with danazol or leuprorelin acetate depot increased sICAM-1 levels (P<0.001); (3) 3 months after termination of both treatments, sICAM-1 levels were unchanged. Although the mechanism leading to the increase of sICAM-1 needs to be further clarified, any benefits of medical treatment of endometriosis such as danazol or leuprorelin appear to be independent of changes in ICAM-1 serum levels.

Adolescent↗
PubMed2001

L-carnitine modifies the humoral immune response in mice after in vitro or in vivo treatment.

Although the role of L-carnitine (L-Cn) as a cofactor in the oxidation of long-chain fatty acids has been well established, this agent has also been recognized to have an important role in the regulation of carbohydrate metabolism, and consequently, the maintenance of cell membrane structure and cell viability. L-Cn has been reported to reduce the apoptotic levels of CD4+ and CD8+ cells. It has also been demonstrated to interfere with cells of the monocytic lineage by regulating their ability to produce growth factors that ultimately affect both T and B lymphocytic subsets. Therefore, in this study, we examined whether this agent affects the antigenic response of immune cells and determined the relative numbers of immune cells in the murine spleen after in vitro and in vivo treatment. The results showed that L-Cn reduces the relative numbers of CD8+, CD4+ and Ly5+ cells. This observation was consistent in all systems studied including (a) in vitro inoculation of antigen (DNP-HSA) and L-Cn, (b) in vitro priming of spleen cells treated with L-Cn in vivo, and (c) in vivo immunization and L-Cn administration. In all cases, the reduction of T lymphocytes correlated with the decreased production of interleukin-2. L-Cn, however, did not affect the production of specific antibody, which indicates that the observed reduction of Ly5-positive cells is due to cell differentiation of B cells to plasma cells.

Animals↗
PubMed2001

Serum-soluble human leucocyte antigen class I and class II concentrations as an alternative diagnostic test for determining immune indices required for normal pregnancies.

The levels of maternal immunostimulation (required throughout the gestation period) and immunosuppression (needed from the 8th week to labour), as assessed by the mixed lymphocyte reaction (MLR), have been successfully correlated with the outcome of pregnancy. Our laboratory has recently reported that serum-soluble human leucocyte antigen (HLA) class I and II concentrations can be predictive for successful pregnancy outcome. In fact, there is a direct correlation between soluble class II concentrations and maternal immunostimulation because, as expected, these serum HLA concentrations are augmented in the first and second trimester of pregnancy and remain stable thereafter. By the same token, serum HLA class I concentrations are low during the first trimester, correlating with the required absence of immunosuppression, whereas they increase in subsequent trimesters as suppression becomes desirable for counteracting the maternal stimulation, which may otherwise become dangerous to the fetus. In this study, we present biological and statistical evidence that both states of maternal immunostimulation and immunosuppression, reflected by serum soluble HLA class II and class I antigens, do correlate with results obtained by standard MLR and can be predictive of pregnancy failure. The establishment of statistically significant correlations renders the measurement of soluble HLA a reliable test for determining the immunological status of the gestating woman. The unambiguous advantage of such an approach is that soluble HLA testing will no longer require the 1 week delay necessary to obtain MLR results, a period occasionally crucial for applying treatment to women whose immunological indices call for immediate therapeutic intervention.

Adult↗
PubMed2001

The possible anti-inflammatory role of circulating human leukocyte antigen levels in women with endometriosis after treatment with danazol and leuprorelin acetate depot.

BACKGROUND: Endometriosis is defined as an inflammatory condition of the female reproductive tract, a state often associated with infertility and miscarriage. Many exogenously administered factors (treatments) control the disease via as yet unknown pathways. Possible candidate molecules involved in these mechanisms could be the serum-soluble human leukocyte antigens (sHIA) that have been detected in a variety of human body fluids and that are associated with several diseases. AIMS: We here examine how danazol and leuprorelin acetate depot treatments exert their anti-inflammatory action. It is plausible that subtle alterations mediated by these treatments and in relation to sHLA may explain the pathophysiology of endometriosis and provide insights towards new therapeutic protocols. METHODS: Indirect enzyme-linked immunosorbent assay (ELISA), using specific monoclonal antibodies, determined serum-soluble class-I and class-II HLA levels. ELISA readings from treated women were compared with normal healthy subjects. RESULTS: Serum-soluble class-I and class-II HLA levels are statistically significantly lower (P < 0.001) in women with endometriosis than in the control groups. However, danazol but not leuprorelin acetate depot administration augments soluble HLA class I and class II (P < 0.01 and P < 0.001, respectively) to normal levels during the treatment period, an increase that may account for the anti-inflammatory effect and the remission observed. CONCLUSIONS: It is shown that one of the underlying causes of endometriosis may be the lack of both circulating class-I and class-II antigen levels. Danazol administration acts via an induced release of these antigens, whose presence correlates with the degree of the inflammatory alleviation obtained. We thus provide evidence that the inflammatory state of the disease appears to be associated with soluble HLA levels because, 3 months after ceasing therapy, the circulating antigens in the serum return to the same levels that correspond to the pathological condition.

Adult↗
PubMed2000

Localization of pepstatin's inhibitory action during Fc-mediated antibody internalization: possible implications for antibody-mediated viral transmission.

Antibody internalization via Fc receptors is an important cellular mechanism, possibly facilitating the entry of antigenic peptides or viral particles into cells when specific antibodies are present at the periphery. Using an experimental model of trophoblast cells, we have shown that anti-p21(ras) monoclonal antibodies can use IFN-gamma-induced surface Fcgamma receptors to enter the cell. This entry of anti-p21(ras) antibodies ultimately inhibits IFN-gamma-mediated class II antigen induction. Since there may be obvious and inevitable harmful aspects of this mechanism, during which Fc-mediated viral particle or autoantigen transport may occur, we concentrated efforts on defining a potent inhibitor able to eliminate such uptake. The results presented here show that the protease inhibitor pepstatin A efficiently inhibits Fcgamma receptor induction by IFN-gamma and also blocks the endocytic pathway followed by an antibody when it enters the cell at the level of early endosomal compartments. We thus postulate that the use of pepstatin A, because of its inhibition of autoantigen presentation or viral transmission, including that of HIV, may find important applications in therapeutic protocols.

Animals↗
PubMed2000

Th1- and Th2-type lymphokine-assisted induction and release of chemokine receptors from primary human trophoblast cells.

Chemokine receptors (CCRs) have been demonstrated to facilitate the entry of HIV in different cell types of infected individuals, including CD4(+) T cells and dendritic cells. The natural or inducible expression of CCRs on trophoblast cells could provide a valid mechanism for the in utero transmission of HIV from mother to fetus. Because of the rapid turnover of these receptors, we attempted to define the natural and inducible expression of surface CCR3 and CXCR4 on primary human trophoblasts during short periods of cell culture. In the absence of any external stimulus the expression of CCR3 and CXCR4 varied from 1% to 24%. Kinetic experiments show that the levels of both CCR3 and CXCR4 reach a peak of expression after 6 h of culture, whereas by 24 h they have almost disappeared. In the presence of IFN-gamma, CCR3 is showing an increasing pattern of expression after 4 h of incubation, reaching highest levels after 24 h of culture, whereas CXCR4 is kept at lower levels as compared with nontreated cells. Furthermore, in the presence of IL-4, CCR3 expression declines from 2 to 8 h of culture to increase again at 24 h, where 50% of the cell population is expressing the receptors. Under the IL-4 stimulus, CXCR4 shows a peak of expression at 8 h of culture. An interesting feature of this study is that we were able to detect soluble CCR activity in the culture supernatants of trophoblast cells, which followed an inverse pattern of this of surface expression. Thus, the inability of many laboratories to detect high levels of CCRs in placentae of HIV infected mothers may be due on these fast turnover of these receptors, which by the assaying time have either been released in the culture medium or been internalized to the cell.

Cells, Cultured↗
PubMed2000

IFN-gamma facilitates release of class II-loaded intracellular pools in trophoblast cells: a novel property independent of protein synthesis.

Interferon-gamma (IFN-gamma) is an abortion-inducing factor, yet its effects in such a reaction are subject to various levels of regulation. The trophoblast cell line TROPHO-1 can be induced by IFN-gamma to express mRNA and surface class II major histocompatibility complex (MHC) proteins after 8 and 48 h of stimulation, respectively. Untreated cells, however, show an intracellular accumulation of class II antigens earlier (6 h), indicating the existence of MHC pools in the cystosol independent of any induction. On addition of IFN-y, immunofluorescence, subcellular fractionation, and ELISA experiments showed that class II antigen activity detected in the endosomal compartments of the cells could be measured in the culture supernatants. These soluble class II proteins, when isolated and purified using magnetic bead isolation techniques and tested in SDS-PAGE gel and Western blot experiments, had a molecular weight of 70 kDa. Administration of these molecules to pregnant mice as culture supernatants increased the abortion rate and decreased maternal hematocrit levels, effects that could be immunoabsorbed by anti-I-A(d) monoclonal antibodies (mAb). These results indicate that although surface class II molecules are not expressed on trophoblast cells, they accumulate in endosomal compartments and can be released from the cells on addition of IFN-gamma. This new IFN-gamma property, to mobilize intracellular pools of class II MHC antigens in trophoblast cells independent of de novo protein synthesis and induce their release to the extracellular matrix, is a mechanism that appears to be involved in the fetal rejection process, facilitating priming of the maternal organism against the fetal allograft.

Abortion, Induced↗
PubMed2000

Nitric oxide production by pre-implantation embryos in response to embryotoxic factors.

In this report, we examined whether nitric oxide (NO) is involved in early embryo death. We have chosen various experimentally defined embryotoxic stimuli in mice and determined their ability to induce NO production by 2-cell stage embryos. The embryotoxic factors used were interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), lipopolysaccharide (LPS), 5-Azacytidine (5-AzaC) and the murine embryotoxic antibody DF4. We showed that in all cases the embryotoxic stimuli induced NO production by early stage embryos that correlated with the induction of the inducible and/or endothelial isoforms of NO synthase. This study was also extended to the human system where sera from women who aborted were tested for their ability to act embryotoxically by inducing NO in early mouse embryos and mature murine placenta. The results obtained confirmed the embryotoxic character of NO found in these particular sera leading to the hypothesis that NO plays a potential role in early embryo death.

Abortion, Spontaneous↗