Using a human microarray to highlight new genes of interest for a better understanding of molecular mechanisms that underpin the physiopathology of heavesRamery, Eve ; ; Bureau, Fabrice et alin Proceedings: Plant and Animal Genome Conference, Equine Workshop, San Diego (2008) Environmental causes of heaves are well described, but the molecular mechanisms of the disease remain unclear. Using reverse transcription polymerase chain reaction (RT-PCR), disparate results have been ... [more ▼] Environmental causes of heaves are well described, but the molecular mechanisms of the disease remain unclear. Using reverse transcription polymerase chain reaction (RT-PCR), disparate results have been obtained concerning cytokines expression profile. cDNA microarray appears to be so far the platform of choice for massively parallel gene expression profiling and provides a good tool for exploratory research. However, equine-specific microarrays are not yet available on the market. Because they are commercially available, highly specific and well annotated, human and mouse large-scale microarrays are an exploratory alternative to equine-specific microarrays. In the present study, the purpose was to highlight new targets not previously related to the disease and able to improve our understanding of the molecular mechanisms of the disease. A human microarray was used to study gene expression in nucleated cells originating from peripheral blood and bronchoalveolar lavage fluid in heaves-affected horses. With a four-fold cut-off, a total of 46 candidates were identified with differentially regulated genes between heaves-affected horses and controls. Based on their documented function, five of these genes were selected for the real-time quantitative RT-PCR (RT-qPCR) validation procedure: CYBB, BTG1, MARCKS, PTX3 and PTPRC. The RT-qPCR results confirmed those obtained with the microarray, pointing out these genes as new directions for future experiments. However, the human microarray failed to detect the presence of IL-1beta and Il-8, otherwise confirmed by RT-qPCR, and the expression profile of the disease could not be obtained [less ▲] Detailed reference viewed: 19 (2 ULg) Investigation of pulmonary expression of heat shock protein HSP70 in mice chronically exposed to cigarette smokeCheu, Esteban ; Steuve, Jonathan ; Fievez, Laurence et alin Acta Physiologica (2008), 192(662), 19 Detailed reference viewed: 53 (13 ULg) Lung interstitial macrophages prevent lipopolysaccharide-triggered T helper type 2 responses to harmless inhaled antigens; Wallemacq, Hugues ; Marichal, Thomas et alin Proceedings of the Annual BIS-meeting (2008) Detailed reference viewed: 23 (4 ULg) Anti-inflammatory effects of ipratropium bromide in rats with cadmium-induced acute pulmonary inflammation; Fievez, Laurence ; Cheu, Esteban et alin Proceedings of the 12th Annual Meeting of the French Society of Pharmacology and Therapeutics (2008) Detailed reference viewed: 19 (2 ULg) How transcriptomic studies may help to improve the control of bovine diseases : an example with calf pneumonia and endotoxemiaWallemacq, Hugues ; Ramery, Eve ; Bureau, Fabrice et alin Proceedings: XXVth Jubilee World Buiatrics Congress (2008) Global change in genes expression induced by pathological processes can now be analysed in cattle by new tools called microarrays. These transcriptomic studies may help to better understand the ... [more ▼] Global change in genes expression induced by pathological processes can now be analysed in cattle by new tools called microarrays. These transcriptomic studies may help to better understand the pathophysiological mechanisms responsible for the disease and may therefore contribute to develop more efficient preventive and curative strategies. An example is given with a model of calf pneumonia and endotoxemia [less ▲] Detailed reference viewed: 40 (9 ULg) Influence of different exposure conditions to cigarette smoke on the pulmonary acute inflammatory response in miceSteuve, Jonathan ; Cheu, Esteban ; Fievez, Laurence et alin Acta Physiologica (2008), 192(suppl. 662), 18 Detailed reference viewed: 42 (13 ULg) Role of beta 2-receptors in the anti-inflammatory effects of formoterol in rats with cadmium-induced acute pulmonary inflammation; Fievez, Laurence ; Cheu, Esteban et alin Fundamental & Clinical Pharmacology (2008), 227 Detailed reference viewed: 33 (8 ULg) Expression génique et médecine vétérinaireRamery, Eve ; ; Bureau, Fabrice et alin 36èmes Journées AVEF (2008) Detailed reference viewed: 40 (16 ULg) Hunting for a key to the enigma of heaves in the black box of the white cellsArt, Tatiana ; Bureau, Fabrice ; in Veterinary Journal (2008), 177(3), 307-308 Detailed reference viewed: 53 (21 ULg) Dendritic cells genetically engineered to express IL-10 induce long-lasting antigen-specific tolerance in experimental asthma.; Desmet, Christophe ; et alin Journal of Immunology (2008), 181(10), 7230-7242 Dendritic cells (DCs) are professional APCs that have a unique capacity to initiate primary immune responses, including tolerogenic responses. We have genetically engineered bone marrow-derived DCs to ... [more ▼] Dendritic cells (DCs) are professional APCs that have a unique capacity to initiate primary immune responses, including tolerogenic responses. We have genetically engineered bone marrow-derived DCs to express the immunosuppressive cytokine IL-10 and tested the ability of these cells to control experimental asthma. A single intratracheal injection of OVA-pulsed IL-10-transduced DCs (OVA-IL-10-DCs) to naive mice before OVA sensitization and challenge prevented all of the cardinal features of airway allergy, namely, eosinophilic airway inflammation, airway hyperreactivity, and production of mucus, Ag-specific Igs, and IL-4. OVA-IL-10-DCs also reversed established experimental asthma and had long-lasting and Ag-specific effects. We furthermore showed, by using IL-10-deficient mice, that host IL-10 is required for mediating the immunomodulatory effects of OVA-IL-10-DCs and demonstrated a significant increase in the percentage of OVA-specific CD4(+)CD25(+)Foxp3(+)IL-10(+) regulatory T cells in the mediastinal lymph nodes of OVA-IL-10-DC-injected mice. Finally, adoptive transfer of CD4(+) mediastinal lymph node T cells from mice injected with OVA-IL-10-DCs protected OVA-sensitized recipients from airway eosinophilia upon OVA provocation. Our study describes a promising strategy to induce long-lasting Ag-specific tolerance in airway allergy. [less ▲] Detailed reference viewed: 54 (15 ULg) Relevance of using a human microarray to study gene expression in heaves-affected horses.Ramery, Eve ; ; Bureau, Fabrice et alin Veterinary Journal (2008), 177(2), 216-221 Environmental causes of heaves are well described, but the molecular mechanisms of the disease remain unclear. Previous studies have highlighted the implications of variations in gene expression, most ... [more ▼] Environmental causes of heaves are well described, but the molecular mechanisms of the disease remain unclear. Previous studies have highlighted the implications of variations in gene expression, most using reverse transcription polymerase chain reaction (RT-PCR). This well-known technique limits the number of genes that can be studied in a single assay. Microarray appears to be a valuable tool to by-pass this limitation, but so far there has been no equine-specific microarray available on the market. The present study was performed to determine whether a human microarray could be used to study gene expression in nucleated cells originating from peripheral blood and bronchoalveolar lavage fluid (BALF) in heaves-affected horses. With a four-fold cut-off, a total of 46 candidates were identified with differentially regulated genes between heaves-affected horses and controls. A real-time quantitative RT-PCR (RT-QPCR) conducted on a selection of genes, determined on the basis of previous publications, was used to validate the microarray results. The microarray failed to detect the presence of interleukin (IL)-1beta and IL-8 mRNA in the nucleated cells from BALF otherwise confirmed by real-time RT-QPCR. Although some candidate genes have been identified using this method, a complete expression profile of genes related to heaves could not be obtained with the use of the human microarray. [less ▲] Detailed reference viewed: 34 (13 ULg) Nicotinamide phosphoribosyl transferase/pre-B cell colony-enhancing factor/visfatin is required for lymphocyte development and cellular resistance to genotoxic stress.; ; et al in Journal of Immunology (2008), 181(7), 4685-4695 Nicotinamide phosphoribosyl transferase (Nampt)/pre-B cell colony-enhancing factor (PBEF)/visfatin is a protein displaying multiple functional properties. Originally described as a cytokine-like protein ... [more ▼] Nicotinamide phosphoribosyl transferase (Nampt)/pre-B cell colony-enhancing factor (PBEF)/visfatin is a protein displaying multiple functional properties. Originally described as a cytokine-like protein able to regulate B cell development, apoptosis, and glucose metabolism, this protein also plays an important role in NAD biosynthesis. To gain insight into its physiological role, we have generated a mouse strain expressing a conditional Nampt allele. Lack of Nampt expression strongly affects development of both T and B lymphocytes. Analysis of hemizygous cells and in vitro cell lines expressing distinct levels of Nampt illustrates the critical role of this protein in regulating intracellular NAD levels. Consequently, a clear relationship was found between intracellular Nampt levels and cell death in response to the genotoxic agent MNNG (N-methyl-N'-nitro-N-nitrosoguanidine), confirming that this enzyme represents a key regulator of cell sensitivity to NAD-consuming stress secondary to poly(ADP-ribose) polymerases overactivation. By using mutant forms of this protein and a well-characterized pharmacological inhibitor (FK866), we unequivocally demonstrate that the ability of the Nampt to regulate cell viability during genotoxic stress requires its enzymatic activity. Collectively, these data demonstrate that Nampt participates in cellular resistance to genotoxic/oxidative stress, and it may confer to cells of the immune system the ability to survive during stressful situations such as inflammation. [less ▲] Detailed reference viewed: 45 (8 ULg) Influence of different exposure conditions to cigarette smoke on the pulmonary acute inflammatory response in mice; Cheu, Esteban ; Fievez, Laurence et alin Acta Physiologica (2008), 192(662), 18 Detailed reference viewed: 18 (4 ULg) Expression génique et médecine vétérinaireRamery, Eve ; Van Erck, Emmanuelle ; Bureau, Fabrice et alin Proceedings des 36èmes Journées AVEF (2008) Detailed reference viewed: 16 (2 ULg) Use of a human microarray to highlight new genes of interest for a better understanding of recurrent airway obstruction in horses (heaves)Ramery, Eve ; ; Bureau, Fabrice et alin XVIth International Plant & Animal Genome Conference (2008) Detailed reference viewed: 29 (11 ULg) Anti-inflammatory effects of ipratropium bromide in rats with cadmium-induced acute pulmonary inflammationZhang, Yinghong ; Fievez, Laurence ; Cheu, Esteban et alin Fundamental & Clinical Pharmacology (2008), 22(1), 7 Detailed reference viewed: 42 (3 ULg) Potential use of micro-array technology (bio-puce) in the diagnosis of inflammatory disorders in the horseLekeux, Pierre ; ; Ramery, Eve et alin XVII. Tagung über Pferdekrankheiten im Rahmen der EQUITANA (2007) Detailed reference viewed: 48 (13 ULg) Stat5 Is an Ambivalent Regulator of Neutrophil HomeostasisFievez, Laurence ; Desmet, Christophe ; et alin PLoS ONE (2007), 2(1), 727 Although STAT5 promotes survival of hematopoietic progenitors, STAT5-/- mice develop mild neutrophilia. METHODOLOGY/PRINCIPAL FINDINGS: Here, we show that in STAT5-/- mice, liver endothelial cells (LECs ... [more ▼] Although STAT5 promotes survival of hematopoietic progenitors, STAT5-/- mice develop mild neutrophilia. METHODOLOGY/PRINCIPAL FINDINGS: Here, we show that in STAT5-/- mice, liver endothelial cells (LECs) autonomously secrete high amounts of G-CSF, allowing myeloid progenitors to overcompensate for their intrinsic survival defect. However, when injected with pro-inflammatory cytokines, mutant mice cannot further increase neutrophil production, display a severe deficiency in peripheral neutrophil survival, and are therefore unable to maintain neutrophil homeostasis. In wild-type mice, inflammatory stimulation induces rapid STAT5 degradation in LECs, G-CSF production by LECs and other cell types, and then sustained mobilization and expansion of long-lived neutrophils. CONCLUSION: We conclude that STAT5 is an ambivalent factor. In cells of the granulocytic lineage, it exerts an antiapoptotic function that is required for maintenance of neutrophil homeostasis, especially during the inflammatory response. In LECs, STAT5 negatively regulates granulopoiesis by directly or indirectly repressing G-CSF expression. Removal of this STAT5-imposed brake contributes to induction of emergency granulopoiesis. [less ▲] Detailed reference viewed: 86 (39 ULg) Prolactin-induced activation of nuclear factor kappa B in bovine mammary epithelial cells: Role in chronic mastitisBoutet, Philippe ; Sulon, Joseph ; et alin Journal of Dairy Science (2007), 90(1), 155-164 We sought to determine whether prolactin (PRL) could influence the neutrophilic inflammation that characterizes chronic mastitis. Most of the genes encoding inflammatory proteins depend on the nuclear ... [more ▼] We sought to determine whether prolactin (PRL) could influence the neutrophilic inflammation that characterizes chronic mastitis. Most of the genes encoding inflammatory proteins depend on the nuclear factor kappa B (NF-kappa B) for their expression. We addressed the hypothesis that immunomodulatory activities of PRL might arise from an increase in NF-kappa B activity. MAC-T cells, a bovine mammary epithelial cell line, were stimulated with increasing concentrations of bovine PRL ( 1, 5, 25, 125, and 1,000 ng/mL). Level of NF-kappa B binding activity was measured and mRNA was evaluated for IL-1 beta, IL-6, IL-8, granulocyte-macrophage colony-stimulating factor (GMCSF), IFN-gamma, and tumor necrosis factor (TNF)-alpha, cytokines known to require NF-kappa B for their maximal transcription. Prolactin activated NF-kappa B; maximal NF-kappa B activation was weaker with PRL than with TNF-alpha at 30 or 180 min poststimulation. In addition, PRL significantly amplified, in a dose-dependent manner, mRNA expression of IL-1 beta, IL-6, IL-8, GMCSF, and TNF-a. We measured PRL concentrations in blood and milk from healthy and chronic mastitis-infected cows, and studied the relationship between the PRL concentration and the degree of inflammation in the mammary gland as indirectly assessed by somatic cell counts (SCC). Plasma PRL did not differ significantly between healthy and chronic mastitis-affected cows (63.7 and 67.5 ng/mL, respectively). Milk PRL concentration was significantly increased in chronic mastitis-affected quarters with the highest SCC, and had a positive significant correlation between SCC, as well as between the number of neutrophils present in milk samples. The present findings show that PRL promotes an inflammatory response in bovine mammary epithelial cells via NF-kappa B activation, and suggest a role for PRL in the pathogenesis of chronic mastitis. [less ▲] Detailed reference viewed: 38 (7 ULg) Anti-inflammatory effects of formoterol in rats after a single dose inhalation of nebulized cadmium; Fievez, Laurence ; Cheu, Esteban et alin Fundamental & Clinical Pharmacology (2007), 74 Detailed reference viewed: 26 (7 ULg) |
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