Biomedical subjects

I Vachier

Publications and source records attributed to I Vachier.

30 records · Page 2Linked to original sources

PubMed1997

[Leukotriene antagonists. A new therapeutic approach in asthma].

BRONCHIAL INFLAMMATION: Corticosteroids are currently the most important treatment for bronchial inflammation in asthma. SULFIDOPEPTIDE LEUKOTRIENES: These biologically active fatty acids play an important role in the inflammation process in asthma. A NEW THERAPEUTIC APPROACH: Some leukotriene receptor antagonists and synthesis inhibitors are effective and safe treatment for mild to moderate asthma. GLOBAL STRATEGY: The role of these new drugs in the management of asthma remains to be defined on the basis of comparative clinical trials with the standard reliever treatment: inhaled corticosteroids.

Asthma↗

PubMed1997

Cellular protein kinase C activity in asthma.

Protein kinase C (PKC) is a primary group of enzymes mediating signal transduction for a wide variety of functions in many different cell types. Its activation has been implicated in various inflammatory diseases. In asthma, inflammatory cells, such as alveolar macrophages (AM) and polymorphonuclear neutrophils (PMN), are primed and activated compared with those obtained from control subjects. In particular, they release higher amounts of reactive oxygen species. PKC has been known to play an important role in the respiratory burst of human leukocytes. In this study, the PKC activity was measured in blood neutrophils and alveolar macrophages from control (n = 16) and asthmatic subjects (n = 28). In PMN, the total PKC activity was significantly lower in PMN from stable (182.00 +/- 27.20) and unstable (108.40 +/- 14.15) asthmatic patients, compared with control subjects (257.35 +/- 29.70 pmol/10(7) cells/min with p < 0.05 and p < 0.0005, respectively). In AM, PKC activity was 479.50 +/- 71.80 for controls and 254.00 +/- 25.90 pmol/10(7) cells/min for asthmatic patients. Moreover the percentage of membrane PKC was significantly higher in stable asthmatic patients in both cell types. After stimulation of neutrophils with PMA, a significant decrease in total PKC activity was observed in both control and asthmatic subjects. We have found an abnormal regulation of PKC activity in both blood PMNs and AM in asthmatic patients. These findings are consistent with the functional hyperreactivity of inflammatory cells observed in asthma.

Adult↗

PubMed1996

5(S),15(S)-dihydroxyeicosatetraenoic acid and lipoxin generation in human polymorphonuclear cells: dual specificity of 5-lipoxygenase towards endogenous and exogenous precursors.

5-Lipoxygenase activation of human blood polymorphonuclear cells (PMN) from asthmatic patients (asthmatics) was studied to investigate whether differences may exist with healthy subjects (controls). The respective cell capacities to produce lipoxins (LXs), leukotrienes, and 5(S), 15(S)-dihydroxyeicosatetraenoic acid [5(S),15(S)-diHETE] were compared under in vitro stimulation by ionophore A23187, with or without exogenous 15(S)-hydroxyeicosatetraenoic acid [15(S)-diHETE]. Eicosanoids were analyzed by elution with an isocratic reverse-phase high performance liquid chromatography system, and their profiles, detected by simultaneous monitoring at 302, 280, and 246 nm, were evaluated on the basis of chromatographic behavior: UV spectral characteristics and coelution with synthetic standards. In the presence of exogenous 15(S)-HETE, human PMN were able to produce LXs and 5(S),15(S)-diHETE, PMN from asthmatics were able to produce 5(S), 5(S),15(S)-diHETE, and LXs from endogenous sources, whereas in the same experimental conditions, no detectable amounts of these compounds were released by PMN from controls. The levels of 5(S),15(S)-diHETE, and LXs biosynthesized from endogenous arachidonic acid were highly correlated. Two different LX patterns were observed involving two possible metabolic pathways: (a) via the intermediate 5,6-epoxytetraene alone for LXs generation from exogenous 15(S)-HETE; and (b) via 5,6- and/or 14,15-epoxytetraenes leading to the formation of an enzyme-bound delocalized carbocation for LXs generation from endogenous arachidonate, respectively. The enhanced 5-lipoxygenase activation of blood PMN from asthmatics and the metabolism of exogenous 15(S)-HETE may reflect a priming induced by various mediators released from environmental cells, and could be considered as a model of transcellular signalization between PMN and endothelial cells.

Arachidonate 5-Lipoxygenase↗

PubMed1996

Glucocorticoids induced down-regulation of glucocorticoid receptor mRNA expression in asthma.

Although their precise mechanism of action remains to be elucidated, glucocorticoids represent the most effective therapy in the treatment of asthma. Interactions between the glucocorticoid receptor and the AP-1 complex have been shown to regulate the transcription of some genes, including glucocorticoid receptor itself. The aim of the present study was to compare the expression of mRNA for glucocorticoid receptor in human blood monocytes obtained from seven unstable untreated asthmatic patients who were subsequently treated with high doses of parenteral corticosteroid (methyl prednisolone 120 mg/day) for 10 days. mRNA expression was identified after RNA extraction using RNAzol and analysed after reverse transcriptase, by polymerase chain reaction using a semiquantitative competitive hybridization assay. All asthmatic patients showed an improvement in their FEV1 values after corticosteroid treatment (per cent of predicted value 68.28 +/- 4.93 versus 95.57 +/- 6.41, P < 0.02), and a significant decrease for glucocorticoid receptor mRNA expression (P < 0.02) was observed in their monocytes. This is the first report of an ex vivo down-regulation for the glucocorticoid receptor mRNA expression, following corticosteroid treatment.

Adult↗

PubMed1995

Nonradioactive quantification of glucocorticoid receptor expression during differentiation of human monocytic cells (U937).

We describe a method for relative quantification of specific mRNA using a nonradioactive assay based on DNA strand competition between identical sequences of biotin- and fluorescein-labeled amplicon (probe) and unlabeled amplicon (target) during hybridization. As the target quantity increased, that of the double-labeled probe decreased in accordance with the mass action law. This technique was successfully applied to evaluate differences in glucocorticoid receptor expression in U937 cells before and after the addition of potent differentiation inducers: 12-O-tetradecanoylphorbol 13-acetate (TPA) and a combination of all-trans retinoic acid (RA) and 1,25-dihydroxyvitamin D2 (VD). We observed that TPA treatment was associated with an increase in specific binding of [3H]dexamethasone and up-regulation of GR mRNA while no enhanced GR expression was perceived with RA/VD treatment.

Base Sequence↗

PubMed1995

5-15-diHETE and lipoxins generated by neutrophils from endogenous arachidonic acid as asthma biomarkers.

5-Lipoxygenase (5-LO) activation of human blood polymorphonuclear cells (PMN) from healthy subjects (HS) and from asthmatic patients (AP) was investigated comparing their respective capacities to produce lipoxins, 5,15-dihydroxyeicosatetraenoic acid (5,15-diHETE) and leukotrienes, under in vitro stimulation by ionophore A23187. PMN from AP were able to generate higher leukotriene levels from endogenous sources than PMN from HS. Moreover they produced 5,15-diHETE (from 50 to 280ng/10(7) cells) and lipoxins (from 1 to 30ng/10(7) cells), in a linear manner, whereas in the same experimental conditions no detectable amounts of these compounds appeared in PMN from HS. The enhanced 5-LO activation of blood PMN may reflect transcellular signalisation priming indicating that lipoxins and 5,15-diHETE could be much more specific inflammatory state biomarkers than leukotriene B4.

Arachidonic Acid↗

PubMed1995

Bimodal action of fatty acids on PMA-stimulated O2.- production in human adherent monocytes.

Not only unsaturated linear fatty acids, but also saturated monomethyl-branched fatty acids (MMBFAs), are provided by food. They are capable of penetrating into the membrane lipids and promoting lipid disorder. Both NADPH oxidase and protein kinase C (PKC) are bound to cell cytoplasmic membranes and are responsive to unesterified fatty acids (UEFAs). We found that the O2.- NADPH oxidase-mediated production of human adherent monocytes was modified by unsaturated linear and saturated branched UEFAs only in the presence of phorbol myristate acetate. This result together with our inhibition data indicated an action strictly linked to PKC activity. The type and intensity of action depended on the UEFA concentrations and chain structure. Nanomolar concentrations showed potentiating effects whereas micromolar (< CMC) concentrations displayed depressant influences MMBFAs were generally more active than the other FAs. With respect to the micromolar depressant effect, oleate and linoleate were as active and docosahexaenoate nearly as active as MMBFAs. As assessed by iso-15:0 or arachidonate action, such bimodal alteration did not occur in non-adherent monocytes and neutrophils. Certain UEFAs could be considered as 'cellular' anti-oxidants on the sites of adherent-monocyte recruitment.

Acridines↗

PubMed1994

IgE produces monocyte superoxide anion release: correlation with CD23 expression. Comparison of patients with asthma, patients with rhinitis, and normal subjects.

Allergic inflammation involves many different cell types among which mononuclear cells, such as macrophages and monocytes, play an important role. These cells release numerous chemical mediators, including superoxide anion (O2.-). We evaluated the capacity of atopic serum to stimulate peripheral blood monocyte O2.- release. Thirteen untreated allergic patients (seven with asthma and six with rhinitis), and five nonallergic control subjects were studied. O2.- was measured in a photon-counting camera with Lucigenin-enhanced (Sigma Chemical Co., St. Louis, Mo.) chemiluminescence. Results were expressed (mean +/- SEM) in relation to basal values (peak/basal chemiluminescence values). Spontaneous production of O2.- was greater in allergic patients. Moreover, atopic serum stimulated O2.- production of blood monocytes in all subjects, but this was greater in subjects with allergic asthma than in subjects with allergic rhinitis and normal subjects. Anti-IgE immunoadsorption of atopic serum completely abrogated this effect, which was restored by the IgE-rich eluted fraction. IgE-induced O2.- release decreased as adherence duration increased and was correlated with surface CD23 expression. These results indicate that monocytes from allergic patients are in an activated state and that binding of IgE to their receptors generates O2.-, possibly by direct activation of blood monocyte reduced nicotinamide adenine dinucleotide phosphate oxidase.

Adult↗

PubMed1994

Enhancement of reactive oxygen species formation in stable and unstable asthmatic patients.

There is increasing evidence to suggest that human blood polymorphonuclear neutrophils (PMNs) and monocytes play an important role in the inflammatory processes of asthma. In asthmatic patients, PMNs and monocytes were shown to be activated more than in healthy subjects. We investigated the capacity of these two cell populations to generate reactive oxygen species (ROS) in stable and unstable asthmatic patients. The two populations of asthmatic patients were identified by asthma activity, as expressed by clinical events occurring within 2 weeks prior to the study. Oxygen species formation was analysed for isolated purified PMNs and monocytes (Mos) by chemiluminescence (CL) using lucigenin and luminol as luminescent probes. CL was determined on nonstimulated and on phorbol myristate acetate (PMA)-stimulated cells. The stimulatability coefficient (PMA-stimulated/nonstimulated cell ratio) of each cell population was then calculated. Resting PMNs and Mos generated significantly greater amounts of ROS in stable asthmatic patients, and much more in unstable asthmatic patients, as compared to healthy subjects, both in lucigenin and luminol enhanced CL. Non O2.- ROS production from PMA-stimulated PMNs and Mos was identical in unstable asthmatic patients and in healthy subjects, whereas a significant decrease was observed in stable asthmatic patients, as assessed by luminol enhanced CL. PMA-stimulated cells showed no difference in O2.- generation, as assessed by lucigenin enhanced CL. However, the stimulatability coefficient of all asthmatic patients was always significantly lower than that of healthy subjects. These results suggest that there are differences in priming and stimulation of Ros production from PMNs and Mos between stable and unstable asthmatic patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Acridines↗

PubMed1992

Increased oxygen species generation in blood monocytes of asthmatic patients.

Besides eosinophils, inflammatory processes in asthma are characterized by an infiltration of inflammatory cells, including mononuclear phagocytes, such as alveolar macrophages (AM) and blood monocytes, in the airways. Monocyte activation has been observed in the blood after exercise or allergen-induced asthma. Stimulated AM in chronic and stable asthmatic patients have been shown to release oxygen species. We thus investigated the intensity of the activation of monocytes from 18 asthmatic patients compared with 18 healthy subjects. Oxygen species release was analyzed for monocytes in suspension by chemiluminescence using a luminometer and for monocytes maintained in adherence using conventional assay and video imaging camera. Circulating blood monocytes in suspension from asthmatic patients and control subjects showed the same baseline free radical release. Monocytes in suspension from asthmatic patients were more stimulatable by PMA: specifically, monocytes release more H2O and peaks of O2-. are sooner; moreover, peaks of total free radical release are higher, and this plateau is sustained. Compared with monocytes from control subjects, those from asthmatic patients evaluated after adherence show a higher baseline for O2-. and higher total free radical release. Monocytes from asthmatic patients spontaneously release more O2-. over time in nonstimulated cells and release more O2-. with PMA stimulation; they show the same peak level total free radical release as those from control subjects after stimulation. SOD activity analysis on adherent monocytes was lower in asthmatic compared with control subjects. These data show that monocytes from asthmatic patients were activated compared with control monocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

Acridines↗

PubMed1990

[Aberrant expression of HLA-DR antigens of the MHC class II in bronchial epithelial cells in asthmatic patients].

The study of cellular events in the bronchi of asthmatic patients shows significant epithelial destruction. The ciliated cells are more often destroyed than others in the respiratory epithelium. Using highly specific monoclonal antibodies, we detected that the epithelial cells were positive for HLA-DR antigen expression, whereas those from healthy subjects were negative. The aberrant expression could be explained by the presence of T lymphocytes in the asthmatic bronchial mucosa. The activated lymphocytes (CD4+) are known to release gamma interferon, which is a lymphokine that induces or enhances the expression of HLA-DR antigens. These cells, which are known to mediate cytotoxicity in an Ag-specific and Ia-restricted way, could take part in the shedding process of epithelial cells in asthma.

Asthma↗

PubMedDate not supplied

Imaging reactive oxygen species in asthma.

Inflammatory processes in asthma are characterized by an infiltration of inflammatory cells including mononuclear phagocytes. It has been observed that mononuclear phagocytes, alveolar macrophages and blood monocytes, release higher quantities of reactive oxygen species in asthmatic patients than in healthy subjects. Chemiluminescence assays were developed to measure the superoxide anion and the other reactive oxygen species. The chemiluminescence response was first analysed with a luminometer, which made it possible to study cells in suspension before and after PMA-stimulation. Secondly a video-imaging camera was used in experiments on adherent cells before and after stimulation with PMA and/or specific stimulus IgE/anti-IgE. Both techniques showed that human alveolar macrophages, blood monocytes, PMN and lymphocytes were spontaneously primed in vivo and were more easily stimulated in asthma. Analysis of adherent cells in vitro may provide give information on the physiological condition of adherent cells in vivo.

Acridines↗