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T Lecompte

Publications and source records attributed to T Lecompte.

At least 19 recordsLinked to original sources
PubMed2001

Effects of three Hb-based oxygen-carrying solutions on neutrophil activation in vitro: quantitative measurement of the expression of adherence receptors.

BACKGROUND: Hb-based oxygen carriers (HbOCs) are currently under investigation as RBC substitutes. These solutions have vasoactive properties and may modify the behavior of the RBC in vitro. The effects of these agents on other blood cells have not been extensively investigated. The aim of this study was to evaluate the potential effect(s) of three differently modified HbOCs on the activation of PMNs. STUDY DESIGN AND METHODS: Whole blood from humans was incubated with Hb dextran-benzene-tetracarboxylate, alphaalpha-Hb, or o-raffinose-poly-Hb for 15, 30, and 60 minutes. The expression of adherence receptors CD62L, CD18, and CD11b, which reflects the activation state of the neutrophils, was assessed in vitro by a direct immunofluorescence method using calibration beads measured by flow cytometry. RESULTS: The observed expression of each of the three adherence receptors for each solution at each time point was similar to that noted with the RPMI control. Decreases in CD62L and increases in CD18 expression were noted at 15 minutes of incubation but, when those values were compared to the values obtained with TNFalpha as the positive control of PMN activation, the profile of expression of adherence receptors obtained with HbOC solutions did not conform to the expected activation profile of PMNs. CONCLUSION: Hb solutions modified by a variety of methods, as obtained and employed in this study, did not appear to activate neutrophils in vitro.

Aspirin↗
PubMed2001

[Cutaneous delayed hypersensitivity reactions to heparins and heparinoids].

INTRODUCTION: Due to cross-reactions between unfractionated heparins, low-molecular-weight heparins and sometimes heparinoids, cutaneous delayed hypersensitivity reactions might be a problem for the choice of therapeutic alternative. We report on two cases of sensitization to heparins and heparinoids. OBSERVATIONS: One woman developed localized skin reaction to a low-molecular-weight heparin, then a generalized maculopapular rash when an intravenous injection of unfractionated heparins was performed. The second patient had a localized then extended reaction to a low-molecular-weight heparin and was referred for the choice of a well tolerated method to obtain anticoagulation during a pulmonary surgery. METHODS: Patch tests, prick tests, intradermal and subcutaneous tests were performed with several unfractionated heparins, low-molecular-weight heparins, danaparoid and lepirudin in both cases. RESULTS: In the first case, tests performed with both heparins and heparinoid were positive and the use of lepirudin was proposed if anticoagulation was necessary. In the second case the subcutaneous danaparoid injection induced a localized reaction on the injection site. Danaparoid injections were continued associated with localized applications of dermocorticoids without any side effect. DISCUSSION: Cutaneous delayed hypersensitivity reactions occur on the injection site but can also be generalised. Cross reactions might be unexpected therefore skin-tests are necessary to guide the choice of a therapeutic alternative. In case of intolerance to both heparins and heparinoids various solutions may be proposed such as the application of topical corticosteroids on the injection site or the administration of hirudins. There is no cross sensitization between heparins and hirudins, but the use of hirudins is restricted and requires specific monitoring.

Drug Eruptions↗
PubMed2001

Beta2-glycoprotein I binding to platelet microparticle membrane specifically reduces immunoreactivity of glycoproteins IIb/IIIa.

We have investigated beta2-glycoprotein I (beta2GPI) binding to platelet-derived microparticles (PMP) and its effect on GPIIb/IIIa. PMP were isolated from washed human platelets after stimulation with A23187, and analyzed by surface plasmon resonance spectroscopy. Beta2GPI as well as activated protein C (APC) or annexin V bound to PMP-coated sensorchips, demonstrating exposure of anionic phospholipids on immobilized PMP. Beta2GPI binding was impaired by calcium and occurred in a concentration-dependent manner with apparent k(on) = 2.6 x 10(4) M(-1) s(-1) and k(off) = 4.4 x 10(-3) s(-1), corresponding to a KD value of 1.7 x 10(-7) M. When analyzed by flow cytometry, the binding of certain mAbs specific for GPIIb and/or GPIIIa was reduced in the presence of beta2GPI but not of APC or annexin V, whereas the binding of anti-GPIb or anti-P-selectin mAbs, or of soluble fibrinogen remained unchanged. These results suggest a broad but specific influence of beta2GPI on GPIIb/IIIa immunoreactivity, and indicate that beta2GPI may act as a modulator of GPIIb/IIIa-dependent functions of PMP.

Antibodies, Monoclonal↗
PubMed2000

Recombinant hirudin anticoagulation for aortic valve replacement in heparin-induced thrombocytopenia.

PURPOSE: To report the case of a patient with HIT that received a prolonged infusion of r-hirudin (lepirudin; Refludan; Hoechst, France) before, during and after cardiopulmonary bypass (CPB) for aortic surgery. Although administration of r-hirudin for CPB anticoagulation has previously been reported, many questions persist concerning the best therapeutic regimen for CPB anticoagulation as well as the time of onset and the doses for postoperative anticoagulation. CLINICAL FEATURES: A 65-yr-old man was admitted for surgery of aortic stenosis after an episode of acute pulmonary edema complicated by deep venous thrombosis in the context of documented HIT. The patient received r-hirudin for 13 dy before surgery at doses (0.4 mg x kg(-1) bolus followed by 0.15 mg x kg(-1) x hr(-1) continuous infusion) that maintained activated partial thromboplastin time (aPTT) ratios between 2 and 2.5. Anticoagulation for CPB was performed with r-hirudin given as 0.1 mg x kg(-1) i.v. bolus and 0.2 mg kg(-1) in the CPB priming volume. Anticoagulation during CPB was monitored with the whole blood activated coagulation time and ecarin clotting time (ECT) performed in the operating room with values corresponding to r-hirudin concentrations >5 microg x ml(-1) during CPB. Anticoagulation during CPB was uneventful. Two bleeding episodes, related to the r-hirudin regimen and necessitating allogeneic blood transfusion, occurred after surgery. CONCLUSION: This case report confirms previous experience of the use of r-hirudin for anticoagulation during CPB and provides additional information in the context of prolonged r-hirudin infusion before and after CPB.

Aged↗
PubMed2000

Monovalent binding of autoantibodies to beta2-glycoprotein I, detected using surface plasmon resonance at low antigen density.

The precise mechanism of interaction between autoantibodies and beta2-glycoprotein I (beta2GPI) and the experimental conditions to be used for their detection are still under debate. Until now, these interactions have been studied under static conditions. We have investigated the interactions of purified IgG from 25 lupus anticoagulant-positive patients with immobilized beta2GPI under flow conditions by real-time analysis based on surface plasmon resonance technology. Sensor chips were coated with purified human beta2GPI coupled to dextran via amino groups at low densities (1.4, 1.8 or 2. 4 ng beta2GPI/mm2). Four patients' IgG displayed efficient binding and had the highest so-called antiphospholipid IgG levels by enzyme-linked immunosorbent assay (ELISA) and the highest absorbance values in an anti- beta2GPI ELISA at a beta2GPI density reported to be around 12 ng/mm2. Binding of antibodies to the beta2GPI sensor chips proved to be dependent upon the IgG concentration and beta2GPI density and was inhibited by a rabbit antibody against beta2GPI. Similar association and dissociation profiles were observed for the four efficient binders. The fast rate of dissociation limited the binding of autoantibodies to beta2GPI and was highly suggestive of a monovalent association, confirmed by binding of Fab fragments under similar experimental conditions. In conclusion, monovalent binding of low-affinity antibodies to beta2GPI immobilized at a density as low as 1.8 ng/mm2 could be detected using surface plasmon resonance.

Animals↗
PubMed2000

Further evidence for the presence of anti-protein S autoantibodies in patients with systemic lupus erythematosus.

Acquired protein S (PS) deficiency in systemic lupus erythematosus (SLE) has been previously reported, but its mechanism and its possible thrombotic role have not been established. The aim of our study was to provide further evidence for auto-immune PS deficiency in 27 Tunisian SLE patients, using PS-specific enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance technology (SPR). PS deficiencies for PS activity, free PS or total PS, respectively, were found in 19, 18 and 12 patients. A significant correlation (r= -0.475, P< 0.016) was found between free/total PS ratio and C4bBP levels, suggesting a role of inflammation in free PS deficiency. Immunoglobulin IgG antibodies to PS were detected in four patients by both ELISA and SPR, in six patients only by ELISA, and in two patients only by SPR. Signals for anti-PS IgG by ELISA and SPR were, however, significantly correlated (r = 0.549, P = 0.003). These results suggest that an auto-immune mechanism could account for low PS activity in patients with SLE. Auto-antibodies to PS may form immune complexes, inducing increased clearance of PS or interfering with the protein C-protein S system.

Autoantibodies↗
PubMed2000

[Determination of polymorphonuclear neutrophil adhesion receptors. Effect of pre-analytic factors].

UNLABELLED: Polymorphonuclear neutrophil (PMN) adherence receptors expression varies with leukocyte activation state. Their quantification need accurate and inter-laboratories reproducible methods, without artefactual activation. OBJECTIVE: The aim of this study was to study the influence of cell preparation on PMN adherence receptors expression. MATERIALS AND METHODS: It was proposed to quantify, using immunolabeling standard (QIFIKIT(R), Dako), surface expression of the main adherence receptors (L-selectin and B(2) integrins), from different preparations of PMN: total blood collected with EDTA, isolated PMN by density gradient Polymorphprep(TM) 1,113 (Nycomed Pharma) and formaldehyde fixed PMN. RESULTS: A decrease of all receptors was noted after isolation and fixation of PMN, in comparison with whole blood PMN analysis. These results differed from data previously reported since, in these studies, activated phenotypes (increased of B(2) integrins) were observed after isolation and fixation methods. CONCLUSION: The present study provides strong evidence that pre-analytical conditions are sources of biological variations and thus extreme care must be taken in the interpretation of results. It underlines the interest of consensual practices for these pre-analytic and analytic parameters in order to compare results in multicenter and longitudinal studies.

CD18 Antigens↗
PubMed1999

[Antiplatelet agents and their therapeutic use].

Antiplatelet agents have a well established effect against thrombosis complicating atherosclerosis. Drugs currently in use in France are: aspirin and flurbiprofen, inhibiting thromboxane synthesis; ticlopidine and clopidogrel, inhibiting platelet activation by adenosine diphosphate; dipyridamole, inhibiting platelet activation through adenosine; abciximab, acting on the mechanism of aggregation. Their molecular and cellular pharmacology is in agreement with the clinical effects and the guidelines for practical use. These drugs have in common that they carry a risk of hemorrhage, albeit low but difficult to cope with in case of invasive procedures. There is no antidote. They also have specific contraindications. No laboratory tests aimed at assessing their effect on primary haemostasis are proved of any clinical value.

Arteriosclerosis↗
PubMed1999

[Improvement in prevention of venous thromboembolism in medical environment. Mission impossible?].

PURPOSE: In France, low molecular weight heparins are widely used for prophylaxis of venous thromboembolism. This practice is not based on strong evidence supporting the use of this technique, thus leading to important difficulties in defining accurate guidelines. Improvement in the prophylaxis must focus on the process leading to prescription or non-prescription. Our primary objective was to identify and describe this process. We also analyzed the causes for dysfunction and implemented corrective actions. METHODS: Basic tools and methods specific to improvement of quality were used including: flow charting for process description, causes-effect diagram for updating and analysis of dysfunction, weighted vote for decisions regarding corrective actions, and agenda and designation of leaders for the implementation of corrective actions. RESULTS: The theoretical process beginning with the recognition of a risk and ending with the adequate treatment or prevention of this risk was demonstrated. The most important dysfunction was the lack of epidemiological and clinical trials. Key corrective actions were the following: standardization of practices where evidence was available; if not, collaborative efforts to collect valid epidemiological data through multicenter surveys and clinical trials; systematic appraisal of the quality of published data; assessment of the risk for venous thromboembolism during hospitalization. To achieve real improvement in practice, priority was given to repeated measures of the risk for thromboembolism. The use of low molecular weight heparins has been so wide however, that it has led to some difficulties. We must draw lessons from this experience. CONCLUSION: Improvement of quality in terms of healthcare is not only based on auditing the practices or modifying the defective processes, but also on our ability not to use new drugs before fully assessing their validity.

Anticoagulants↗
PubMed1999

Both kinetic data and epitope mapping provide clues for understanding the anti-coagulant effect of five murine monoclonal antibodies to human beta2-glycoprotein I.

The interaction between five murine monoclonal antibodies (mAb) and beta2-glycoprotein I (beta2GPI) in the absence of phospholipids was studied using surface plasmon resonance-based biosensor technology. Two separate epitope regions were confirmed for the five mAb but epitopes of two mAb were shown to be overlapping but not identical. The characteristics of binding on both immobilized beta2GPI, using different chemistries of coupling to a dextran matrix and antibody surfaces prepared by two strategies of immobilization, were compared. Binding was strongly influenced by the orientation of the immobilized partner, and the five mAb showed heterogeneity in their binding to immobilized and soluble beta2GPI. The observed stoichiometries of mAb-beta2GPI complexes and the detailed analysis of the kinetics of the association and dissociation phases of the interactions with soluble and immobilized beta2GPI revealed differences in the dissociation rate constants, resulting in a 10-fold higher affinity for immobilized beta2GPI compared to soluble beta2GPI for four out of five mAb. This suggests bivalent binding of these mAb to immobilized beta2GPI. In addition, the kinetic data helped explain the differing anti-coagulant properties of these mAb.

Animals↗
PubMed1999

[Vascular disease and cellular activation: exploration of cell adhesion phenotype by quantitative flow cytometry].

The pathogenesis of atheromatous and/or thrombotic vascular diseases involves rheological parameters, soluble mediators and cellular agents. The many studies that have tried to establish correlations between plasma factors, shear stress and the risk of ischemia have left some questions unanswered. Current exploration methods are now focusing on the determining role of cells. Activated cells express adhesion molecules on their membranes, which allow to communicate in a homo- or heterotypical manner. Quantifying adherence molecules on the surface of platelets, leukocytes and endothelial cells provides an assessment of the "adhesive phenotypical profile". Quantitative cytometry, using beads coated with a known amount of immunoglobulins as calibrators, is perfectly suited, through its multiple parameter analyses and the specificity provided by monoclonal antibodies, for the quantification of membrane antigens. Measuring the adhesive profile on the surface of cells that are implicated in vascular disease makes it possible to correlate that phenotype to the ischemic risk in such diversified pathologies or circumstances as intermittent angor, myocardial infarction, angioplasty, insulin-dependent diabetes or pre-eclampsia. In addition, that quantification permits monitoring the action of new therapeutical agents targeting adherence molecules.

Angioplasty, Balloon, Coronary↗
PubMed1999

[New thrombolytic agents in myocardial infarction].

Myocardial infarction is the result of thrombotic coronary artery occlusion. Although present-day thrombolytics have major value by increasing the frequency of reopening of arteries responsible for myocardial infarction, by preserving myocardial function and, thereby, significantly reduce mortality. Nevertheless, they are subject to the following limitations: 1) excellent arterial partency is only obtained in 50% of cases: 2) reocclusion occurs in 5 to 10% of cases; 3) severe complications such as cerebral haemorrhage are observed in about 0.5% of cases. Therefore, the search to improve thrombolytic agents is intense. This article reports the recent advances in concept and production of new thrombolytic agents. The most recent results concern the production of mutants of T-PA (tissue plasmogen activator). Of these mutants, the reteplase (r-PA) has already received authorization for its commercialisation. Other t-PA mutants under development (phase 3) include TNK-t-PA and lanoteplase. Over the last few years, there has been renewed interest in staphylokinase. The results of the initial clinical trials with this agent have also been reported. Paradoxically, the mode of action of thrombolytic agents has an inherent pro-thrombotic effect. This explains some of the interest for anti-thrombotic agents as an adjuvant treatment of thrombolysis. The initial results of the association of thrombolytics with new glycoprotein IIb/IIIa platelet inhibitors and anti-thrombin agents are reported.

Coronary Disease↗
PubMed1999

Heparin-induced thrombocytopenia in France, 1980-1998.

Due to the extensive use of unfractionated heparins in France, there is considerable experience with heparin-induced thrombocytopenia (HIT). It is recommended that platelet counts be performed twice a week for three weeks when patients are treated with any form of heparin. A drop in platelet counts can, however, occur not only in HIT patients but also for other unrelated reasons. For diagnosing HIT, all laboratories in France use platelet aggregometry inspite of poor sensitivity. Both false positive and false negative results are obtained. The serotonin release test is not used in France. The ELISA test for HIT does not always correlate with the platelet aggregation test and many patients with a positive ELISA test do not necessarily have other evidence for HIT. This is especially true in patients following cardiopulmonary bypass surgery. None of the available laboratory tests reliably identify patients with HIT. Patients with HIT should not be managed with low-molecular-weight heparins, but danaparoid, argatroban and ancrod are viable options. Also, recombinant hirudin has been employed. All have advantages and disadvantages. At present, the diagnosis and management of patients with HIT remains difficult and properly designed clinical studies are needed to obtain answers to several open questions.

Anticoagulants↗
PubMed1998

Immunopurification of human beta2-glycorprotein I with a monoclonal antibody selected for its binding kinetics using a surface plasmon resonance biosensor.

The beta2-glycoprotein I (beta2GPI)-binding properties of five murine monoclonal antibodies immobilized as capture antibodies were studied using surface plasmon resonance detection. The monoclonal antibody with the fastest dissociation kinetics (6F3) was selected for the development of an immunoaffinity chromatography procedure, assuming that its behaviour would be similar in both systems since the covalent coupling chemistries involved amino groups in both cases. Under our experimental conditions of a fast one-step procedure, beta2GPI was purified to homogeneity from human plasma with a yield of about 50%. Beta2GPI was eluted under fairly mild conditions, either at low pH or at high pH. The immunoadsorbent was used five times without any apparent loss of binding capacity. The immunopurified protein showed similar binding to cardiolipin-coated polystyrene wells as beta2GPI purified by conventional methods. However, differences in the pattern of immunoreactivity in relation to the purification procedure were observed by surface plasmon resonance using the monoclonal antibody with the highest association kinetics (9G1) immobilized on the sensor surface.

Animals↗
PubMed1998

[Prevention of venous thromboembolism. Survey of in-hospital medical practice].

OBJECTIVES: In an effort to improve the prevention of venous thromboembolism, the Nancy University Hospitals conducted a survey of medical practice concerning indications for preventive therapy and surveillance of platelet counts and anti Xa activity. METHODS: The survey involved 163 medical files. Questionnaires were filled out in 6 units (3 medical wards and 3 intensive care units). RESULTS: Indications for preventive therapy were found to be quite variable with the exception of very low risk of thromboembolism where the treat/do not treat ratio was 0.1/1, indicating a clear tendency for abstention. This ratio was 0.77/1 and 0.38/1 respectively for low and moderate risk and 2/1 for high risk. There was undoubtedly a ward effect. The attitudes in practice tended toward non-prevention in patients without limited mobility. For platelet counts, an initial count was performed in 95% of the cases and during treatment in 38% although the specific rates were not the same for different types of units. Anti-Xa activity, which according to prevention recommendations need not to be determined, was not monitored in 88% of the cases. In accordance with prevention recommendations, anti-Xa activity was not determined in 88% of the cases. CONCLUSION: Further progress is needed in the prevention of venous thromboembolism and should be based on wider use of existing methods.

France↗