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Biomedical subjects

F Dignat-George

Publications and source records attributed to F Dignat-George.

At least 37 records · Page 2Linked to original sources
PubMed2000

Neutrophil activation in preeclampsia and isolated intrauterine growth restriction.

OBJECTIVE: Neutrophils have been implicated in the pathogenesis of preeclampsia. Because preeclampsia and intrauterine growth restriction result from similar placental lesions, the aim of this study was to investigate neutrophil activation in isolated intrauterine growth restriction relative to preeclampsia and uncomplicated pregnancy. Polymorphonuclear neutrophil activation was analyzed by measuring cell surface and soluble cell adhesion molecule expressions. STUDY DESIGN: L -Selectin (CD62L ) and CD11b surface expressions on polymorphonuclear neutrophils were analyzed in 13 women with preeclampsia, 11 women with isolated intrauterine growth restriction, and 17 age- and gestation-matched control women by means of a standardized quantitative flow cytometry assay. Serum levels of soluble L -selectin were measured by enzyme-linked immunosorbent assay. RESULTS: Neutrophils from women with isolated intrauterine growth restriction and women with preeclampsia displayed higher levels of CD11b and lower levels of CD62L than did neutrophils from healthy pregnant women. Soluble L -selectin serum levels were significantly increased in the preeclampsia and intrauterine growth restriction groups relative to normal values. No significant difference in the levels of CD11b, CD62L, and soluble L -selectin were observed between women with isolated intrauterine growth restriction and those with preeclampsia. Leukocyte activation was not correlated with disease severity. CONCLUSION: The observed alteration in polymorphonuclear neutrophil adhesion molecule expressions and increased serum soluble L -selectin levels are consistent with activation of peripheral blood neutrophils occurring in isolated intrauterine growth restriction in a manner similar to that seen in preeclampsia. This evidence of neutrophil activation may help to advance our understanding of the disease process in isolated intrauterine growth restriction.

Adult↗
PubMed2000

Profile of endothelial and leukocyte activation in Fabry patients.

Fabry disease is an X-linked recessive disorder resulting in the deposition of globotriaosylceramide in numerous cell types including vascular endothelial cells. Because this disease is associated with vascular injury and a high recurrence rate of thrombotic events, measurements of factors regulating endothelium and leukocyte interaction may provide insight into the mechanisms leading to a prothrombotic state. Twenty-five patients with Fabry disease and 25 control subjects participated in the study. Plasma from all 25 Fabry patients and 15 of the 25 controls were studied for multiple endothelial factors. Leukocyte integrins were measured by flow cytometry in 21 Fabry patients and 10 controls. The concentrations of soluble intercellular adhesion molecule-1, vascular cell adhesion molecule-1, P-selectin, and plasminogen activator inhibitor were significantly higher and thrombomodulin was significantly lower in Fabry patients. Expression of the integrin CD11b on monocytes was also significantly higher in the Fabry patients. This study reveals measurable evidence for endothelium and leukocyte activation that is consistent with a prothrombotic state in Fabry patients compared with controls. Further investigations of these findings may help to understand the mechanism of stroke in Fabry disease and provide indicators (or markers) of efficacy of future therapeutic intervention.

Adult↗
PubMed1999

Direct evidence of endothelial injury in acute myocardial infarction and unstable angina by demonstration of circulating endothelial cells.

Circulating endothelial cells (CECs) have been detected in association with endothelial injury and therefore represent proof of serious damage to the vascular tree. Our aim was to investigate, using the technique of immunomagnetic separation, whether the pathological events in unstable angina (UA) or acute myocardial infarction (AMI) could cause desquamation of endothelial cells in circulating blood compared with effort angina (EA) and noncoronary chest pain. A high CEC count was found in AMI (median, 7.5 cells/mL; interquartile range, 4.1 to 43.5, P <.01 analysis of variance [ANOVA]) and UA (4.5; 0.75 to 13.25 cells/mL, P <.01) within 12 hours after chest pain as compared with controls (0; 0 to 0 cells/mL) and stable angina (0; 0 to 0 cells/mL). CEC levels in serial samples peaked at 15.5 (2.7 to 39) cells/mL 18 to 24 hours after AMI (P <.05 repeated measures ANOVA), but fell steadily after UA. Regardless of acute coronary events, the isolated cells displayed morphologic and immunologic features of vascular endothelium. The CECs were predominantly of macrovascular origin. They did not express the activation markers intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, and E-selectin, although some were positive for tissue factor. CECs failed to exhibit characteristics of apoptosis (TUNEL assay) excluding this event as a possible mechanism of cell detachment. The presence of CECs provides direct evidence of endothelial injury in AMI and UA, but not in stable angina, confirming that these diseases have different etiopathogenic mechanisms.

Aged↗
PubMed1999

Differential regulation of killer cell Ig-like receptors and CD94 lectin-like dimers on NK and T lymphocytes from HIV-1-infected individuals.

NK and T lymphocytes share various cell surface receptors, including NK receptors for MHC class I molecules (NKR). NKR include killer cell Ig-like receptors (KIR) and lectin-like dimers which are composed of the invariant CD94 associated with a variety of NKG2 molecules. The combination of KIR and CD94/NKG2 dimers expressed on NK and T cell subsets defines a repertoire of MHC class I recognition. Engagement of NKR by cognate MHC class I molecules governs T and NK cell activation. We investigated the NKR distribution on NK and T cell subsets from uninfected and HIV-infected individuals, according to the clinical status, the absolute numbers of CD4+ T cells as well as the plasmatic viral load of the patients. We show that the KIR distribution on NK cells is not affected by HIV-1 infection, whereas the absolute numbers of T cells expressing specific KIR members (CD158b, p70) transiently increase in early stages of HIV infection. By contrast, the percentages of NK and T cells which express CD94 dimers increase in parallel with the disease. These results document a differential regulation of KIR and CD94 lectin-like dimers during the course of a chronic viral infection in humans and further suggest that both types of NKR are independently regulated.

Acquired Immunodeficiency Syndrome↗
PubMed1999

Cholesterol and omega-3 fatty acids inhibit Na, K-ATPase activity in human endothelial cells.

We have investigated the effects of cholesterol and omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexenoic acid (DHA) on Na, K-ATPase activity in human endothelial cells (HUVEC). Cultured HUVEC were incubated for 18 h with pure egg phosphatidylcholine (PC), or cholesterol-enriched liposomes (4 mg PC/ml). EPA and DHA alpha-tocopherol-acetate were emulsified with PC and incubated with HUVEC (10 mM). Na, K-ATPase and 5'-nucleotidase activities were determined using the coupled assay method on microsomal fractions obtained from cultured cells using non treated cells as control. Cholesterol enrichment significantly reduced both Na, K-ATPase and 5'-nucleotidase activities by a similar level (- 40%), whereas pure phospholipid liposomes inhibited this activity only by 22%. The dose-response curves of Na, K-ATPase activity were all biphasic assuming the presence of two independent sites exhibiting different affinities for ouabain of nM and microM respectively. The cholesterol induced inhibitory effect was greater for low affinity sites (-54%) as compared to that of the high affinity sites (-24%) whereas omega-3 fatty acids reduced the activity of both sites by 22%. Short term effects of EPA and DHA on Na, K-ATPase activity were determined by incubating microsomal fractions from untreated cells with various concentrations of free fatty acids (from 1 to 200 microM) for 20 min. Both EPA and DHA significantly reduced Na, K-ATPase activity but inhibition by EPA seems to be more effective than DHA. These results suggest that cholesterol and omega-3 fatty acids reduce Na, K-ATPase activity in HUVEC.

Cell Membrane↗
PubMed1999

In vitro generation of endothelial microparticles and possible prothrombotic activity in patients with lupus anticoagulant.

Microparticles (MPs) resulting from vesiculation of platelets and other blood cells have been extensively documented in vitro and have been found in increased numbers in several vascular diseases, but little is known about MPs of endothelial origin. The aim of this study was to analyze morphological, immunological, and functional characteristics of MPs derived from human umbilical vein endothelial cells (HUVECs) stimulated by TNF, and to investigate whether these MPs are detectable in healthy individuals and in patients with a prothrombotic coagulation abnormality. Electron microscopy evidenced bleb formation on the membrane of TNF-stimulated HUVECs, leading to increased numbers of MPs released in the supernatant. These endothelial microparticles (EMPs) expressed the same antigenic determinants as the corresponding cell surface, both in resting and activated conditions. MPs derived from TNF-stimulated cells induced coagulation in vitro, via a tissue factor/factor VII-dependent pathway. The expression of E-selectin, ICAM-1, alphavbeta3, and PECAM-1 suggests that MPs have an adhesion potential in addition to their procoagulant activity. In patients, labeling with alphavbeta3 was selected to discriminate EMPs from those of other origins. We provide evidence that endothelial-derived MPs are detectable in normal human blood and are increased in patients with a coagulation abnormality characterized by the presence of lupus anticoagulant. Thus, MPs can be induced by TNF in vitro, and may participate in vivo in the dissemination of proadhesive and procoagulant activities in thrombotic disorders.

Antiphospholipid Syndrome↗
PubMed1999

Differences in levels of soluble E-selectin and VCAM-1 in malignant versus non malignant Mediterranean spotted fever.

In the present study, we investigated the plasma levels of soluble adhesion molecules E-selectin, P-selectin, intercellular adhesion molecule- (ICAM- ) and vascular cell adhesion molecule-1(VCAM-1) in 24 patients with Mediterranean spotted fever (MSF), 6 of whom with a malignant form. Measurements were performed on blood samples collected before treatment (T1), then twice during treatment (T2 and T3). Before treatment, MSF patients taken as a whole presented elevated levels of sICAM-1 and sVCAM-1 and normal levels of sE-selectin and sP-selectin compared to healthy controls. We found that sICAM-1 was elevated both in mild and malignant MSF. sE-selectin and sVCAM-1 were elevated only in patients with the malignant form and allowed to discriminate the two clinical subgroups. Their levels decreased after treatment with sE-selectin reaching control values at T2 whereas sVCAM-1 remained higher over the course of the malignant form. In patients with mild MSF, sP-selectin steadily increased after treatment, whereas it did not present any modification at any of the two sampling times in patients with the malignant form. Raised plasma levels of sE-selectin and sVCAM-1 reflect endothelial activation in malignant rickettsial disease and may be sufficiently early markers to influence the therapeutic decision.

Adult↗
PubMed1998

Activation of human endothelial cells via S-endo-1 antigen (CD146) stimulates the tyrosine phosphorylation of focal adhesion kinase p125(FAK).

S-Endo-1 antigen (CD146), a transmembrane receptor also known as MUC18/MCAM, is a member of the immunoglobulin superfamily and belongs to a group of cell adhesion molecules. CD146 is highly expressed on the whole vascular tree. We demonstrate here that engagement of CD146 on human endothelial cells isolated from cord blood results in tyrosine phosphorylation of a large panel of cellular proteins, although no tyrosine phosphorylation of CD146 was detected. In particular, CD146 cross-linking induces the tyrosine phosphorylation of the protein tyrosine kinase p125(FAK) as well as p125(FAK) association with paxillin, both events being inhibited by cytochalasin D. No direct association of CD146 with p125(FAK) was observed. Consistent with these data, CD146 associates with p59(fyn), a Src family kinase known to phosphorylate p125(FAK). The identification of a signaling pathway initiated by CD146 engagement and which includes p59(fyn), p125(FAK), and paxillin indicates that CD146 participates in outside-in signaling in endothelial cells.

Antigens, CD↗
PubMed1998

CD146: biosynthesis and production of a soluble form in human cultured endothelial cells.

We previously identified the S-Endo 1-associated antigen (CD146), an endothelial member of the immunoglobulin superfamily with a characteristic V-V-C2-C2-C2 Ig domain structure. In cultured human endothelial cells, we investigated its biosynthesis by immunoprecipitation and pulse-chase labeling. CD146 was synthesized as a 100 kDa precursor form, which was processed into a 120 kDa mature form. In the culture media of endothelial cells, we observed a CD146 soluble form that was about 10 kDa smaller than cell-associated CD146. In parallel with soluble forms of other members of the immunoglobulin superfamily, soluble CD146 could modulate and control the functions of the molecule.

Antigens, CD↗
PubMed1998

Effect of uremia and hemodialysis on soluble L-selectin and leukocyte surface CD11b and L-selectin.

Leukocyte Mac-1 (CD11b/CD18) and L-selectin (CD62L) are implicated in leukocyte adhesion to endothelial cells. In this study, L-selectin and CD11b expression on leukocytes and soluble L-selectin (sL-selectin) serum levels were investigated in 17 nondialyzed patients with chronic renal failure (CRF), in 28 chronic hemodialysis patients before hemodialysis (basal state), and in 32 healthy subjects. These parameters were also monitored during hemodialysis with cuprophane and cellulose diacetate membranes in a crossover study in five patients. Granulocytes from CRF patients displayed lower expression of L-selectin and higher expression of CD11b than granulocytes from healthy subjects. On the other hand, baseline expression of L-selectin and CD11b on leukocytes from hemodialysis patients did not differ from that of healthy subjects. In CRF and hemodialysis patients, sL-selectin levels were significantly lower than in healthy subjects. During hemodialysis, cuprophane membrane induced an upregulation of granulocyte CD11b, a decrease in granulocyte L-selectin, and an increase in sL-selectin serum levels. Conversely, cellulose diacetate caused only a transient increase in granulocyte CD11b and did not modify granulocyte L-selectin and sL-selectin serum levels. High CD11b and low L-selectin expression on granulocytes in CRF patients suggests an activation state, which was not found in hemodialysis patients at the basal state. The lack of activation in hemodialysis patients could reflect the elimination of a uremic toxin by dialysis or a loss of granulocyte responsiveness because of the repetitive stimulation by hemodialysis treatment. The low serum levels of sL-selectin in CRF and hemodialysis patients also suggest granulocyte dysfunction.

Biocompatible Materials↗
PubMed1998

Evidence that human endothelial cells express different isoforms of Na,K-ATPase.

BACKGROUND: The catalytic alpha and smaller beta subunits of the plasma membrane Na,K-ATPase occur in various molecular forms (alpha1, alpha2, alpha3, beta1 and beta2). The alpha isoforms of the enzyme have varying affinities for ouabain and exist in different tissues with particular distribution patterns. OBJECTIVE: To document the existence of isoforms of the Na,K-ATPase in cultured human umbilical vein endothelial cells. METHODS: Microsomal fractions were prepared by differential ultracentrifugation from primary cultures of human umbilical vein endothelial cells and from such cells obtained after three passages. Na,K-ATPase activity was assayed using the coupled assay method and sensitivity to ouabain was determined in the presence of varying concentrations of ouabain. Specific antibodies for the various Na,K-ATPase isoforms were used to label these different proteins by immunocytochemistry in endothelial cells and by Western blotting in isolated membranes. RESULTS: In plotting the dose-response curves for Na,K-ATPase activity in response to ouabain we assumed the existence of two independent sites exhibiting different affinities for ouabain (in the micromol/l and the nmol/l ranges). The contribution of low-affinity sites was threefold that of high-affinity sites. After three passages in culture, a specific increase in Na,K-ATPase activity of the high-affinity sites was observed compared with that of cells from primary cultures. Confocal microscopy revealed the existence of beta1, beta2, and alpha1 subunit proteins in human umbilical endothelial cells. Staining for alpha3 isoform was less pronounced and no obvious alpha2 was detected. CONCLUSION: These findings suggest that human umbilical vein endothelial cells contain beta1, beta2, a large amount of alpha1 isoform with an apparently low affinity for ouabain, and a lesser amount of high-affinity sites, which may correspond to the alpha3 protein.

Cells, Cultured↗
PubMed1998

Evidence for the expression of urokinase-type plasminogen activator by human venous endothelial cells in vivo.

Endothelial cells (ECs) in culture synthesize and secrete urokinase-type plasminogen activator (u-PA), but the normal vascular endothelium is believed to synthesize only tissue plasminogen activator (t-PA), which is thought to be responsible for intravascular fibrinolysis. More recently, animal studies have shown that the biological role of u-PA in fibrinolysis has been underestimated, prompting a re-examination of its synthesis by the endothelium. In this study, we investigated whether u-PA was synthesized by non-atherosclerotic endothelial cells in vivo by testing ECs dislodged by venipuncture from 12 normal volunteers and 17 patients admitted for plasmapheresis. The ECs were isolated with an anti-endothelial monoclonal antibody coupled to immunomagnetic beads and characterized by morphology and by labelling for vWF, CD31, and UEA-1 binding. U-PA antigen was found in 50% of the ECs from the normal subjects and in 60% of those from patients. U-PA enzymatic activity on zymograms was detected in 50% of the normal samples and 60% of the patient samples, with the latter being more frequently and more strongly positive. U-PA mRNA was found in all the normal and patient samples tested. The results indicate that u-PA is synthesized by the venous endothelium in vivo but that its expression is highly variable.

Adult↗
PubMed1998

A case report: CD8 expression in B-cell chronic lymphocytic leukemia (B-CLL). Prognostic significance of the aberrant CD8 expression.

The human CD8 molecule is usually expressed on T lymphocytes or natural killer cells but not on normal B cells. Over a course of 5 years, we followed a case of B-CLL which aberrantly expressed the CD8 molecule. During this period, the clinical and hematological conditions of the patient were stable. This B-CLL presented a typical immunophenotype (HLA DR+, CD19+, CD5+, CD23+) with monotypic expression of surface immunoglobulin light chain kappa. We confirmed the CD8 expression on these leukemia cells by using double labelling and different antibodies directed against this antigen. We measured the quantitative expression of the CD8 molecule. The number of CD8 molecules per cell was clearly lower on these malignant B cells than on normal T lymphocytes. In order to evaluate the prognostic significance of this CD8 expression, we quantitated in parallel other markers such as CD5, CD23, CD22, CD11c. During 5 years, this aberrant CD8 expression persisted and was associated with an increase of the CD23 and a decrease of CD22 levels, known to correlate with a good prognosis in agreement with the karyotype analysis. Altogether our results led us to conclude that the aberrant CD8 expression in this case of B-CLL may correlate with a non-aggressive form of lymphoproliferative disorder.

CD8 Antigens↗
PubMed1997

[Endothelium and infections].

Endothelial cells control vascular homeostasis by regulating haemostasis, inflammatory and immune response, vascular tone or angiogenesis. The endothelium is the target of infectious agents capable of altering this equilibrium. Such alterations may result from a direct cytopathic effect with disruption of endothelial integrity, or from an indirect effect due to unregulated activation leading to vascular dysfunction. The endothelium is both the target and the effector of human vascular diseases associated with infection such as atherosclerosis, vasculitis, neuropaludism or graft rejection. A better comprehension of endothelial alterations is these diseases should have biological and therapeutic implications.

Arteriosclerosis↗
PubMed1997

Rickettsia conorii infection enhances vascular cell adhesion molecule-1- and intercellular adhesion molecule-1-dependent mononuclear cell adherence to endothelial cells.

Leukocyte adherence to the endothelium is an essential component of the inflammatory response during rickettsial infection. In vitro, Rickettsia conorii infection of endothelial cells enhances the expression of adhesive molecules E-selectin, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in a time- and dose-dependent manner. Rickettsial lipopolysaccharide does not seem to be involved, because polymyxin B does not reduce their expression. The intracellular presence of the organism and de novo host protein synthesis are required for expression of cell adhesive molecules, since rickettsial inactivation by formol and pretreatment of cells with cycloheximide inhibits an increase in expression. The contribution of interleukin-1alpha (IL-1alpha) to this endothelial adhesive phenotype was shown by inhibitory experiments 8 and 24 h after infection with IL-1 receptor antagonist and IL-1alpha blocking antibodies. Enhanced adherence of mononuclear cells to infected endothelial cells involved VCAM-1- and ICAM-1-dependent mechanisms at the late phase of the inflammatory response. This endothelial adhesive phenotype may constitute a key pathophysiologic mechanism in R. conorii-induced vascular injury.

Animals↗