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12. Hematopoietic niche drives FLT3-ITD acute myeloid leukemia resistance to quizartinib via STAT5-and hypoxia-dependent upregulation of AXL

Author

Dumas, Pierre-Yves, Naudin, Cécile, Martin-Lannerée, Séverine, Izac, Brigitte, Casetti, Luana, Mansier, Olivier, Rousseau, Benoît, Artus, Alexandre, Dufossée, Mélody, Giese, Alban, Dubus, Pierre, Pigneux, Arnaud, Praloran, Vincent, Bidet, Audrey, Villacreces, Arnaud, Guitart, Amélie, Milpied, Noël, Kosmider, Olivier, Vigon, Isabelle, Desplat, Vanessa, Dusanter-Fourt, Isabelle, Pasquet, Jean-Max, CHU Bordeaux, Service d'Hématologie Clinique et Thérapie cellulaire, F-33000, Bordeaux., Biothérapies des maladies génétiques et cancers, Université Bordeaux Segalen - Bordeaux 2-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Paris, Institut Cochin, Institut National de la Santé et de la Recherche Médicale INSERM U1016, Centre National de la Recherche Scientifique CNRS UMR8104, F-75014 Paris., Service de Biologie des Tumeurs and Laboratoire d'Hématologie Biologique, Centre Hospitalo-Universitaire CHU Bordeaux, F-33000, Bordeaux., Service Commun des Animaleries, Animalerie A2, Université de Bordeaux, Bordeaux., TBM-Core [Bordeaux] (CNRS UMS 3427 - INSERM US 005), Université de Bordeaux (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Service d'Hématologie Biologique, Assistance Publique-Hôpitaux de Paris, Hôpitaux Universitaires Paris Centre, Paris, France., Université de Paris, Institut Cochin, Institut National de la Santé et de la Recherche Médicale INSERM U1016, Centre National de la Recherche Scientifique CNRS UMR8104, F-75014 Paris isabelle.dusanter@inserm.fr., and Vigon, Isabelle

Subjects
Acute Myeloid Leukemia, [SDV.MHEP.HEM] Life Sciences [q-bio]/Human health and pathology/Hematology, Phenylurea Compounds, [SDV]Life Sciences [q-bio], Hematopoietic Stem Cell Transplantation, [SDV.CAN]Life Sciences [q-bio]/Cancer, [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology, [SDV.BC.IC] Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Article, Up-Regulation, [SDV] Life Sciences [q-bio], Leukemia, Myeloid, Acute, fms-Like Tyrosine Kinase 3, [SDV.CAN] Life Sciences [q-bio]/Cancer, hemic and lymphatic diseases, embryonic structures, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], STAT5 Transcription Factor, Tumor Microenvironment, Humans, Benzothiazoles, Hypoxia, ComputingMilieux_MISCELLANEOUS
Abstract

Internal tandem duplication in Fms-like tyrosine kinase 3 (FLT3-ITD) is the most frequent mutation observed in acute myeloid leukemia (AML) and correlates with poor prognosis. FLT3 tyrosine kinase inhibitors are promising for targeted therapy. Here, we investigated mechanisms dampening the response to the FLT3 inhibitor quizartinib, which is specific to the hematopoietic niche. Using AML primary samples and cell lines, we demonstrate that convergent signals from the hematopoietic microenvironment drive FLT3-ITD cell resistance to quizartinib through the expression and activation of the tyrosine kinase receptor AXL. Indeed, cytokines sustained phosphorylation of the transcription factor STAT5 in quizartinib-treated cells, which enhanced AXL expression by direct binding of a conserved motif in its genomic sequence. Likewise, hypoxia, another well-known hematopoietic niche hallmark, also enhanced AXL expression. Finally, in a xenograft mouse model, inhibition of AXL significantly increased the response of FLT3-ITD cells to quizartinib exclusively within a bone marrow environment. These data highlight a new bypass mechanism specific to the hematopoietic niche that hampers the response to quizartinib through combined upregulation of AXL activity. Targeting this signaling offers the prospect of a new therapy to eradicate resistant FLT3-ITD leukemic cells hidden within their specific microenvironment, thereby preventing relapses from FLT3-ITD clones.

Published
2019
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18. STAT5-and hypoxia-dependent upregulation of AXL

Author

DUMAS, Pierre-Yves, NAUDIN, Cécile, MARTIN-LANNERÉE, Séverine, IZAC, Brigitte, CASETTI, Luana, MANSIER, Olivier, ROUSSEAU, Benoît, ARTUS, Alexandre, DUFOSSÉE, Mélody, GIESE, Alban, DUBUS, Pierre, PIGNEUX, Arnaud, PRALORAN, Vincent, BIDET, Audrey, VILLACRECES, Arnaud, GUITART, Amélie, MILPIED, Noël, KOSMIDER, Olivier, VIGON, Isabelle, DESPLAT, Vanessa, DUSANTER-FOURT, Isabelle, and PASQUET, Jean-Max

Subjects
Article RECHERCHE
Published
2019

19. The Expression of Myeloproliferative Neoplasm-Associated Calreticulin Variants Depends on the Functionality of ER-Associated Degradation

Author

Mansier, Olivier, primary, Prouzet-Mauléon, Valérie, additional, Jégou, Gwénaële, additional, Barroso, Kim, additional, Raymundo, Diana Pelizzari, additional, Chauveau, Aurélie, additional, Dumas, Pierre-Yves, additional, Lagarde, Valérie, additional, Turcq, Béatrice, additional, Pasquet, Jean-Max, additional, Viallard, Jean-François, additional, James, Chloé, additional, Praloran, Vincent, additional, Voutetakis, Konstantinos, additional, Chatziioannou, Aristotelis, additional, Mahon, François-Xavier, additional, Chevet, Eric, additional, and Lippert, Eric, additional

Published
2019
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26. Additional file 3: of MiR-10a and HOXB4 are overexpressed in atypical myeloproliferative neoplasms

Author

Pierre-Yves Dumas, Mansier, Olivier, Prouzet-Mauleon, Valerie, Koya, Junji, Villacreces, Arnaud, Grange, Philippe Brunet De La, Paz, Damien Luque, Bidet, Audrey, Jean-Max Pasquet, Praloran, Vincent, Salin, Franck, Kurokawa, Mineo, François-Xavier Mahon, Cardinaud, Bruno, and Lippert, Eric

Abstract

Figure S1. (A) Expression of HOXB4 in OCI-AML3 according to treatment by 5-aza-2’deoxycytidine (DEOX), valproic acid (VPA), and retinoic acid (RA). OCI-AML3 were treated with 2 μM DEOX and/or 1 mM VPA for 48 h and/or 2 μM RA for 4 h. qRT-PCR measured the relative expression of HOXB4 with TUBA1C and RPLP0 both used as control genes. The graphs show fold induction of gene expression by treatment over DMSO control with mean ± SEM (n = 3). *, p

Published
2018
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28. Additional file 2: of MiR-10a and HOXB4 are overexpressed in atypical myeloproliferative neoplasms

Author

Pierre-Yves Dumas, Mansier, Olivier, Prouzet-Mauleon, Valerie, Koya, Junji, Villacreces, Arnaud, Grange, Philippe Brunet De La, Paz, Damien Luque, Bidet, Audrey, Jean-Max Pasquet, Praloran, Vincent, Salin, Franck, Kurokawa, Mineo, François-Xavier Mahon, Cardinaud, Bruno, and Lippert, Eric

Abstract

Table S1. Characteristics of patients with atypical myeloproliferative neoplasms (n = 18). Table S2. Characteristics of patients with hematological malignancies (n = 39). (DOC 136 kb)

Published
2018
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31. Steady state peripheral blood provides cells with functional and metabolic characteristics of real hematopoietic stem cells

Author

Bourdieu, Antonin, primary, Avalon, Maryse, additional, Lapostolle, Véronique, additional, Ismail, Sadek, additional, Mombled, Margaux, additional, Debeissat, Christelle, additional, Guérinet, Marianne, additional, Duchez, Pascale, additional, Chevaleyre, Jean, additional, Vlaski‐Lafarge, Marija, additional, Villacreces, Arnaud, additional, Praloran, Vincent, additional, Ivanovic, Zoran, additional, and Brunet de la Grange, Philippe, additional

Published
2017
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32. Apolipoprotein A1: A new serum marker correlated to JAK2 V617F proportion at diagnosis in patients with polycythemia vera

Author

François Girodon, Jérôme Garin, Sylvie Hermouet, Praloran Vincent, Jean-Yves Cahn, Marie Arlotto, Pascal Mossuz, François Berger, Sabine Brugière, Ali Bouamrani, Irène Dobo, Eric Lippert, and François Laporte

Subjects
Mutation, biology, Chemistry, Clinical Biochemistry, medicine.disease_cause, medicine.disease, Blood proteins, Real-time polymerase chain reaction, Polycythemia vera, hemic and lymphatic diseases, Immunology, medicine, biology.protein, Biomarker (medicine), Apolipoprotein A1, Allele, JAK2 V617F
Abstract

Polycythemia vera (PV) is a myeloproliferative disorder (MPD) characterized by an acquired gain-of-function mutation of the JAK2 protein (JAK2 V617F). Allele-specific quantitative PCR has showed a JAK2 V617F dosage effect on haematological and clinical parameters of PV at diagnosis, but it is unknown whether the level of certain serum proteins might correlate with the proportion of mutated JAK2. Taking into account that such proteins could represent useful prognostic marker, we investigated the serum protein profile of PV patients by SELDI-TOF MS. We identified apolipoprotein A1 (Apo-A1) as a serum marker correlated to the percentage of JAK2 V617F alleles; Apo-A1 expression being the highest for PV patients with more than 75% of mutated alleles. Immuno-assay on an automated random immuno-analyser confirmed the correlation between Apo-A1 concentrations and JAK2 V617F percentages, and showed that serum Apo-A1 assay allowed the specific discrimination of PV patients with high levels of mutated alleles (≥75%). These data suggest that Apo-A1 assay could be a useful assay for the stratification of PV patients at diagnosis.

Published
2007
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33. Hypoxia/Hypercapnia-Induced Adaptation Maintains Functional Capacity of Cord Blood Stem and Progenitor Cells at 4 degrees C

Author

Vlaški, Marija, Negroni, Luc, Kovačević-Filipović, Milica, Guibert, Christelle, de la Grange, Philippe Brunet, Rossignol, Rodrigue, Chevaleyre, Jean, Duchez, Pascale, Lafarge, Xavier, Praloran, Vincent, Schmitter, Jean-Marie, and Ivanović, Zoran

Abstract

We analyzed the effect of exposure to hypoxic/hypercapnic (HH) gas mixture (5% O2/9% CO2) on the maintenance of functional cord blood CD34+ hematopoietic stem and progenitor cells in severe hypothermia (4 degrees C) employing the physiological and proteomic approaches. Ten-day exposure to HH maintained the Day 0 (D-0) level of hematopoietic stem cells as detected in vivo on the basis of hematopoietic repopulation of immunodeficient miceshort-term scid repopulating cells (SRC). Conversely, in the atmospheric air (20% O2/0.05% CO2), usual condition used for cell storage at 4 degrees C, stem cell activity was significantly decreased. Also, HH doubled the survival of CD34+ cells and committed progenitors (CFCs) with respect to the atmospheric air (60% vs. 30%, respectively). Improved cell maintenance in HH was associated with higher proportion of aldehyde dehydrogenase (ALDH) positive cells. Cell-protective effects are associated with an improved maintenance of the plasma and mitochondrial membrane potential and with a conversion to the glycolytic energetic state. We also showed that HH decreased apoptosis, despite a sustained ROS production and a drop of ATP amount per viable cell. The proteomic study revealed that the global protein content was better preserved in HH. This analysis identified: (i) proteins sensitive or insensitive to hypothermia irrespective of the gas phase, and (ii) proteins related to the HH cell-protective effect. Among them are some protein families known to be implicated in the prolonged survival of hibernating animals in hypothermia. These findings suggest a way to optimize short-term cell conservation without freezing. J. Cell. Physiol. 229: 2153-2165, 2014.

Published
2014

35. γδ T Cells Confer Protection against Murine Cytomegalovirus (MCMV).

Author

Khairallah, Camille, Netzer, Sonia, Villacreces, Arnaud, Juzan, Marina, Rousseau, Benoît, Dulanto Magallanes, Sara Adriana, Giese, Alban, Costet, Pierre, Praloran, Vincent, Moreau, Jean François, Dubus, Pierre, Vermijlen, David, Dechanet-Merville, Julie, Capone, Myriam, Khairallah, Camille, Netzer, Sonia, Villacreces, Arnaud, Juzan, Marina, Rousseau, Benoît, Dulanto Magallanes, Sara Adriana, Giese, Alban, Costet, Pierre, Praloran, Vincent, Moreau, Jean François, Dubus, Pierre, Vermijlen, David, Dechanet-Merville, Julie, and Capone, Myriam

Abstract

Cytomegalovirus (CMV) is a leading infectious cause of morbidity in immune-compromised patients. γδ T cells have been involved in the response to CMV but their role in protection has not been firmly established and their dependency on other lymphocytes has not been addressed. Using C57BL/6 αβ and/or γδ T cell-deficient mice, we here show that γδ T cells are as competent as αβ T cells to protect mice from CMV-induced death. γδ T cell-mediated protection involved control of viral load and prevented organ damage. γδ T cell recovery by bone marrow transplant or adoptive transfer experiments rescued CD3ε-/- mice from CMV-induced death confirming the protective antiviral role of γδ T cells. As observed in humans, different γδ T cell subsets were induced upon CMV challenge, which differentiated into effector memory cells. This response was observed in the liver and lungs and implicated both CD27+ and CD27- γδ T cells. NK cells were the largely preponderant producers of IFNγ and cytotoxic granules throughout the infection, suggesting that the protective role of γδ T cells did not principally rely on either of these two functions. Finally, γδ T cells were strikingly sufficient to fully protect Rag-/-γc-/- mice from death, demonstrating that they can act in the absence of B and NK cells. Altogether our results uncover an autonomous protective antiviral function of γδ T cells, and open new perspectives for the characterization of a non classical mode of action which should foster the design of new γδ T cell based therapies, especially useful in αβ T cell compromised patients., info:eu-repo/semantics/published

Published
2015

36. γδ T Cells Confer Protection against Murine Cytomegalovirus (MCMV)

Author

Khairallah, Camille, primary, Netzer, Sonia, additional, Villacreces, Arnaud, additional, Juzan, Marina, additional, Rousseau, Benoît, additional, Dulanto, Sara, additional, Giese, Alban, additional, Costet, Pierre, additional, Praloran, Vincent, additional, Moreau, Jean-François, additional, Dubus, Pierre, additional, Vermijlen, David, additional, Déchanet-Merville, Julie, additional, and Capone, Myriam, additional

Published
2015
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37. Proteomic study of the impact of the JAK2-V617F mutation on the phenotype of essential thrombocythemia

Author

Pascal Mossuz, François Girodon, Sylvie Hermouet, A Bouamrani, François Berger, Eric Lippert, Praloran Vincent, Irène Dobo, Marie Arlotto, Jean-Yves Cahn, Laboratoire d'hématologie cellulaire et moléculaire (DBPC), CHU Grenoble, Grenoble Institut des Neurosciences (GIN), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d'hématologie, CHU Bordeaux [Bordeaux], Laboratoire d'Hématologie, Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Centre Hospitalier Universitaire d'Angers (CHU Angers), PRES Université Nantes Angers Le Mans (UNAM)-PRES Université Nantes Angers Le Mans (UNAM), Département de cancérologie et d'hématologie, CHU Grenoble-Hôpital Michallon, and Issartel, Jean-Paul

Subjects
Male, Proteomics, Cancer Research, Apolipoprotein B, Proteome, medicine.disease_cause, MESH: Polycythemia Vera, MESH: Janus Kinase 2, 0302 clinical medicine, Polycythemia vera, hemic and lymphatic diseases, MESH: Blood Proteins, Polycythemia Vera, 0303 health sciences, Mutation, biology, MESH: Proteomics, Hematology, Blood Proteins, Phenotype, MESH: Amino Acid Substitution, MESH: Proteome, 030220 oncology & carcinogenesis, Female, JAK2 V617F, Thrombocythemia, Essential, MESH: Genetic Diseases, Inborn, Mutation, Missense, MESH: Phenotype, 03 medical and health sciences, Genetics, medicine, Humans, Allele, Molecular Biology, 030304 developmental biology, MESH: Mutation, Missense, MESH: Humans, Essential thrombocythemia, Genetic Diseases, Inborn, Cell Biology, Janus Kinase 2, medicine.disease, Molecular biology, MESH: Male, Amino Acid Substitution, Serum proteome, Immunology, biology.protein, MESH: Thrombocythemia, Essential, MESH: Female
Abstract

International audience; OBJECTIVE: Previous studies have suggested that two subtypes of essential thrombocythemia (ET) could be separated on the basis of their JAK2 status (V617F-positive or V617F-negative), with a continuum between V617F-positive ET and polycythemia vera (PV). Nevertheless, increasingly contradictory data on the impact of JAK2-V617F (presence and load) on ET phenotype highlight the need for further investigations. MATERIALS AND METHODS: We investigated the influence of JAK2-V617F on ET phenotype using mass spectrometry-based analysis of serum protein profiles of ET patients, comparatively with PV patients. RESULTS: V617F-positive ET and PV displayed significant differences in their serum protein profiles. Furthermore, JAK2-V617F presence did not impact significantly the serum proteome of ET patients: we observed very few differences in serum protein profiles of V617F-positive and -negative ET. Reciprocally, clustering of ET patients on the basis of their serum protein profiles did not correlate with JAK2-V617F presence. Finally, the JAK2-V617F load did not influence serum apolipoprotein A-1 levels in ET, a previously validated marker of JAK2-V617F allele burden in PV. CONCLUSION: Serum proteome of ET patients was not influenced by the presence of JAK2-V617F or by high V617F allelic ratio (up to 50%) suggesting that ET phenotype is, at best, only partially influenced by the JAK2-V617F mutation.

Published
2008
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39. Oxygen concentration influences mRNA processing and expression of the CD34 gene

Author

Brunet De La Grange, Philippe, Barthe, Christophe, Lippert, Eric, Hermitte, Francis, Belloc, Francis, Lacombe, Francis, Ivanovic, Zoran, Praloran, Vincent, Institut Cochin (UMR_S567 / UMR 8104), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Génomique et Physiologie du Poisson (G.P.P.), Université Sciences et Technologies - Bordeaux 1, Laboratoire d'hématologie, CHU Bordeaux [Bordeaux], Composantes innées de la réponse immunitaire et différenciation (CIRID), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Transfert de Genes a Visee Therapeutique Dans les Cellules Souches, Université Bordeaux Segalen - Bordeaux 2-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Transfusion Sanguine Aquitaine-Limousin (CTS AQUITAINE-LIMOUSIN), Centre de Transfusion Sanguine, Muniesa, Annie, Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Université Bordeaux Segalen - Bordeaux 2 - Centre National de la Recherche Scientifique (CNRS), and Université Bordeaux Segalen - Bordeaux 2 - Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], [SDV.OT] Life Sciences [q-bio]/Other [q-bio.OT]
Published
2006

40. Préservation des cellules souches hématopoïétiques (CHS) en hypoxie par le VEGF

Author

Debeissat, Christelle, Brunet de La Grange, Philippe, Ivanovic, Zoran, Praloran, Vincent, Muniesa, Annie, Composantes innées de la réponse immunitaire et différenciation (CIRID), Université Bordeaux Segalen - Bordeaux 2 - Centre National de la Recherche Scientifique (CNRS), Institut Cochin (UMR_S567 / UMR 8104), Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Centre de Transfusion Sanguine Aquitaine-Limousin (CTS AQUITAINE-LIMOUSIN), Centre de Transfusion Sanguine, Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)

Subjects
[SDV.MHEP.HEM] Life Sciences [q-bio]/Human health and pathology/Hematology, [SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], [SDV.OT] Life Sciences [q-bio]/Other [q-bio.OT], [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology
Published
2006

41. Steady state peripheral blood provides cells with functional and metabolic characteristics of real hematopoietic stem cells.

Author

Bourdieu, Antonin, Avalon, Maryse, Lapostolle, Véronique, Ismail, Sadek, Mombled, Margaux, Debeissat, Christelle, Guérinet, Marianne, Duchez, Pascale, Chevaleyre, Jean, Vlaski-Lafarge, Marija, Villacreces, Arnaud, Praloran, Vincent, Ivanovic, Zoran, and de la Grange, Philippe Brunet

Subjects
HEMATOPOIETIC stem cells, CORD blood, CELL proliferation, XENOGRAFTS, HYPOXIA-inducible factor 1
Abstract

Hematopoietic stem cells (HSCs), which are located in the bone marrow, also circulate in cord and peripheral blood. Despite high availability, HSCs from steady state peripheral blood (SSPB) are little known and not used for research or cell therapy. We thus aimed to characterize and select HSCs from SSPB by a direct approach with a view to delineating their main functional and metabolic properties and the mechanisms responsible for their maintenance. We chose to work on Side Population (SP) cells which are highly enriched in HSCs in mouse, human bone marrow, and cord blood. However, no SP cells from SSBP have as yet been characterized. Here we showed that SP cells from SSPB exhibited a higher proliferative capacity and generated more clonogenic progenitors than non-SP cells in vitro. Furthermore, xenotransplantation studies on immunodeficient mice demonstrated that SP cells are up to 45 times more enriched in cells with engraftment capacity than non-SP cells. From a cell regulation point of view, we showed that SP activity depended on O2 concentrations close to those found in HSC niches, an effect which is dependent on both hypoxia-induced factors HIF-1α and HIF-2α. Moreover SP cells displayed a reduced mitochondrial mass and, in particular, a lower mitochondrial activity compared to non-SP cells, while they exhibited a similar level of glucose incorporation. These results provided evidence that SP cells from SSPB displayed properties of very primitive cells and HSC, thus rendering them an interesting model for research and cell therapy. [ABSTRACT FROM AUTHOR]

Published
2018
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44. Hypoxia/Hypercapnia-Induced Adaptation Maintains Functional Capacity of Cord Blood Stem and Progenitor Cells at 4°C

Author

Vlaski, Marija, primary, Negroni, Luc, additional, Kovacevic-Filipovic, Milica, additional, Guibert, Christelle, additional, de la Grange, Philippe Brunet, additional, Rossignol, Rodrigue, additional, Chevaleyre, Jean, additional, Duchez, Pascale, additional, Lafarge, Xavier, additional, Praloran, Vincent, additional, Schmitter, Jean-Marie, additional, and Ivanovic, Zoran, additional

Published
2014
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45. Increased soluble and platelet-associated CD40 ligand in essential thrombocythemia and reactive thrombocytosis

Author

Viallard, Jean-François, Solanilla, Anne, Gauthier, Bruno, Contin, Cécile, Déchanet, Julie, Grosset, Christophe, Moreau, Jean-François, Praloran, Vincent, Nurden, Paquita, Pellegrin, Jean-Luc, Nurden, Alan, Ripoche, Jean, Viallard, Jean-François, Contin, Cécile, Déchanet, Julie, Moreau, Jean-François, Médecine Interne, CHU Bordeaux [Bordeaux]-Hôpital Haut-Lévêque [CHU Bordeaux], CHU Bordeaux [Bordeaux], Fibrose hépatique et cancer du foie, Université Bordeaux Segalen - Bordeaux 2-IFR66-Institut National de la Santé et de la Recherche Médicale (INSERM), Composantes innées de la réponse immunitaire et différenciation (CIRID), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Biothérapies des maladies génétiques et cancers, Université Bordeaux Segalen - Bordeaux 2-Institut National de la Santé et de la Recherche Médicale (INSERM), Service d'hématologie, and Grosset, Christophe

Subjects
Adult, Blood Platelets, Male, medicine.medical_specialty, Platelet associated, T-Lymphocytes, Immunology, CD40 Ligand, Biochemistry, White People, Reactive thrombocytosis, Internal medicine, medicine, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Humans, Platelet, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Platelet activation, Thrombopoiesis, Aged, Thrombocytosis, CD40, biology, Chemistry, Essential thrombocythemia, Platelet Count, Cell Membrane, hemic and immune systems, Cell Biology, Hematology, Middle Aged, medicine.disease, Platelet Activation, Endocrinology, biology.protein, Regression Analysis, Female, France, Thrombocythemia, Essential
Abstract

International audience; CD40 ligand (CD40L) is expressed on activated CD4(+) T lymphocytes and at the activated platelet surface. A circulating soluble form of CD40L (sCD40L) is generated by the way of a proteolytic cleavage. We measured sCD40L in the plasma of either healthy subjects; patients with inflammatory disorders and low, normal, or high platelet count (reactive thrombocytosis); or patients with essential thrombocythemia (ET). A tight correlation was found between the platelet count and plasma sCD40L. ET patients had the highest levels of sCD40L. Platelet-associated CD40L was increased in ET and reactive thrombocytosis, conditions associated with increased platelet regeneration. Platelet-associated CD40L was released upon platelet activation. In conclusion, platelets appear as a reservoir of CD40L that may be a major contributor to circulating sCD40L. Platelet-associated CD40L may be a potential marker of platelet regeneration.

Published
2002
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46. CD34+cells obtained from ''good mobilizers'' are more activated and exhibit lower ex vivo expansion efficiency than their counterparts from ''poor mobilizers''

Author

Ivanović, Zoran, Kovačević-Filipović, Milica, Jeanne, Michel, Ardilouze, Leslie, Bertot, Anne, Szyporta, Milene, Hermitte, Francis, Lafarge, Xavier, Duchez, Pascale, Vlaški, Marija, Milpied, Noel, Pavlović, Mirjana, Praloran, Vincent, Boiron, Jean-Michel, Ivanović, Zoran, Kovačević-Filipović, Milica, Jeanne, Michel, Ardilouze, Leslie, Bertot, Anne, Szyporta, Milene, Hermitte, Francis, Lafarge, Xavier, Duchez, Pascale, Vlaški, Marija, Milpied, Noel, Pavlović, Mirjana, Praloran, Vincent, and Boiron, Jean-Michel

Abstract

BACKGROUND: The classification of patients into good or poor mobilizers is based on CD34+ cell count in their peripheral blood (PB) after granulocyte-colony-stimulating factor (G-CSF) injection. We hypothesized that, apart from their mobilization from marrow to the blood, the response to G-CSF of CD34+ cells also includes activation of proliferation, metabolic activity, and proliferative capacity. STUDY DESIGN AND METHODS: Mobilized PB CD34+ cells purified from samples obtained by cytapheresis of multiple myeloma or non-Hodgkins lymphoma patients of both good (> 50 CD34+ cells/mu L) and poor (< 50 CD34+ cells/mu L) mobilizers were studied. The initial cell cycle state of CD34+ cells after selection and their kinetics of activation (exit from G(0) phase) during ex vivo culture were analyzed. Their proliferative capacity was estimated on the basis of ex vivo generation of total cells, CD34+ cells, and colony-forming cells (CFCs), in a standardized expansion culture. Indirect insight in metabolic activity was obtained on the basis of their survival (viability and apoptosis follow-up) during the 7-day-long conservation in hypothermia (4 degrees C) in the air or in atmosphere containing 3% O-2/6% CO2. RESULTS: CD34+ cells obtained from good mobilizers were in lower proportion in the G(0) phase, their activation in a cytokine-stimulated culture was accelerated, and they exhibited a lower ex vivo expansion efficiency than those from poor mobilizers. The resistance to hypothermia of good immobilizers CD34+ cells is impaired. CONCLUSION: A good response to G-CSF mobilization treatment is associated with a higher degree of proliferative and metabolic activation of mobilized CD34+ cells with a decrease in their expansion capacity.

Published
2010

47. Low O-2 concentrations enhance the positive effect of IL-17 on the maintenance of erythroid progenitors during co-culture of CD34+and mesenchymal stem cells

Author

Krstić, Aleksandra, Krstić, Aleksandra, Vlaški, Marija, Hammoud, Mohammad, Chevaleyre, Jean, Duchez, Pascale, Jovčić, Gordana, Bugarski, Diana, Milenković, Pavle B., Bourin, Philippe, Boiron, Jean-Michel, Praloran, Vincent, Ivanović, Zoran, Krstić, Aleksandra, Krstić, Aleksandra, Vlaški, Marija, Hammoud, Mohammad, Chevaleyre, Jean, Duchez, Pascale, Jovčić, Gordana, Bugarski, Diana, Milenković, Pavle B., Bourin, Philippe, Boiron, Jean-Michel, Praloran, Vincent, and Ivanović, Zoran

Abstract

Co-culture of haematopoietic cells with a stromal cell layer does not mimic the physiological, micro-environmental niche, whose major feature is a low oxygen (O-2) concentration. Thus, in order to study the effects of IL-17 in a context which better approximates the physiological state, we investigated its effects on cell expansion, colony-forming ability, and the phenotypical profile of normal, human blood CD34(+) cells co-cultured for five days with MSC layers at various O-2 concentrations (20%, 12.5% and 3% O-2). We demonstrated that IL-17 enhances CD34(+) and total CFC production during the five days of MSC/CD34(+) co-culture. This effect depends upon the O-2 concentration, reaching its maximum at 3% O-2, and is more pronounced on erythroid progenitors (BFU-E). In addition, the stimulation of IL-6 production by IL-17 in MSC cultures and co-cultures is enhanced by low O-2 concentration. The expression of some differentiation markers (CD34, CD13 and CD41) on haematopoietic cells in co-cultures also depends upon the oxygen concentration. Our results strengthen the concept that physiological levels of O-2 (mistakenly called hypoxia), should be considered as an important environmental factor that significantly influences cytokine activity.

Published
2009

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