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Article ; Online: Phenotyping of tumor infiltrating immune cells using mass-cytometry (CyTOF).

Maby, Pauline / Corneau, Aurélien / Galon, Jérôme

2019 Volume 632, Page(s) 339–368

Abstract: The anti-tumor immune response plays a crucial role in cancer patient outcome as well as in response to the growing family of immunotherapeutic treatments. Improving patient prognostic and therapeutic management requires a better knowledge of the tumor ... ...

Abstract	The anti-tumor immune response plays a crucial role in cancer patient outcome as well as in response to the growing family of immunotherapeutic treatments. Improving patient prognostic and therapeutic management requires a better knowledge of the tumor microenvironment, for which a deep characterization of tumor-infiltrating immune populations is instrumental. Mass cytometry represents an excellent tool in tumor Immunology, as it allows the simultaneous analysis of >40 markers on single cells. In this chapter, we review challenging technical aspects of the mass cytometry phenotyping of tumor infiltrating immune cells, focusing on fresh human solid tumor samples. We first explain how to design mass cytometry experiments, then provide detailed protocols to isolate mononuclear immune cells from solid tissues and to stain them for an acquisition on a mass cytometer. We also discuss how to optimize the preparation of single immune cell samples, and how to ensure the reproducibility of data generated from distinct fresh human samples. Finally, we provide troubleshooting suggestions for difficult sample acquisitions on a mass cytometer.
MeSH term(s)	Animals ; Flow Cytometry/methods ; Humans ; Immunophenotyping/methods ; Lymphocytes, Tumor-Infiltrating/immunology ; Mass Spectrometry/methods ; Neoplasms/immunology ; Single-Cell Analysis/methods ; Tumor Microenvironment
Language	English
Publishing date	2019-10-18
Publishing country	United States
Document type	Journal Article ; Review
ISSN	1557-7988 ; 0076-6879
ISSN (online)	1557-7988
ISSN	0076-6879
DOI	10.1016/bs.mie.2019.07.025
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Article ; Online: Progressive and Coordinated Mobilization of the Skeletal Muscle Niche throughout Tissue Repair Revealed by Single-Cell Proteomic Analysis.

Borok, Matthew / Didier, Nathalie / Gattazzo, Francesca / Ozturk, Teoman / Corneau, Aurelien / Rouard, Helene / Relaix, Frederic

Cells

2021 Volume 10, Issue 4

Abstract: ... ...

Abstract	Background
MeSH term(s)	Animals ; Cell Lineage ; Mice ; Muscle Development ; Muscle, Skeletal/pathology ; Muscle, Skeletal/physiopathology ; Proteome/metabolism ; Proteomics ; Regeneration ; Single-Cell Analysis ; Stem Cells/cytology ; Wound Healing
Chemical Substances	Proteome
Language	English
Publishing date	2021-03-28
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells10040744
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Article ; Online: Comprehensive analysis of early T cell responses to acute Zika Virus infection during the first epidemic in Bahia, Brazil.

Samri, Assia / Bandeira, Antonio Carlos / Gois, Luana Leandro / Silva, Carlos Gustavo Regis / Rousseau, Alice / Corneau, Aurelien / Tarantino, Nadine / Maucourant, Christopher / Queiroz, Gabriel Andrade Nonato / Vieillard, Vincent / Yssel, Hans / Campos, Gubio Soares / Sardi, Silvia / Autran, Brigitte / Rios Grassi, Maria Fernanda

PloS one

2024 Volume 19, Issue 5, Page(s) e0302684

Abstract: Background: In most cases, Zika virus (ZIKV) causes a self-limited acute illness in adults, characterized by mild clinical symptoms that resolve within a few days. Immune responses, both innate and adaptive, play a central role in controlling and ... ...

Abstract	Background: In most cases, Zika virus (ZIKV) causes a self-limited acute illness in adults, characterized by mild clinical symptoms that resolve within a few days. Immune responses, both innate and adaptive, play a central role in controlling and eliminating virus-infected cells during the early stages of infection. Aim: To test the hypothesis that circulating T cells exhibit phenotypic and functional activation characteristics during the viremic phase of ZIKV infection. Methods: A comprehensive analysis using mass cytometry was performed on peripheral blood mononuclear cells obtained from patients with acute ZIKV infection (as confirmed by RT-PCR) and compared with that from healthy donors (HD). The frequency of IFN-γ-producing T cells in response to peptide pools covering immunogenic regions of structural and nonstructural ZIKV proteins was quantified using an ELISpot assay. Results: Circulating CD4+ and CD8+ T lymphocytes from ZIKV-infected patients expressed higher levels of IFN-γ and pSTAT-5, as well as cell surface markers associated with proliferation (Ki-67), activation ((HLA-DR, CD38) or exhaustion (PD1 and CTLA-4), compared to those from HD. Activation of CD4+ and CD8+ memory T cell subsets, including Transitional Memory T Cells (TTM), Effector Memory T cells (TEM), and Effector Memory T cells Re-expressing CD45RA (TEMRA), was prominent among CD4+ T cell subset of ZIKV-infected patients and was associated with increased levels of IFN-γ, pSTAT-5, Ki-67, CTLA-4, and PD1, as compared to HD. Additionally, approximately 30% of ZIKV-infected patients exhibited a T cell response primarily directed against the ZIKV NS5 protein. Conclusion: Circulating T lymphocytes spontaneously produce IFN-γ and express elevated levels of pSTAT-5 during the early phase of ZIKV infection whereas recognition of ZIKV antigen results in the generation of virus-specific IFN-γ-producing T cells.
MeSH term(s)	Humans ; Zika Virus Infection/immunology ; Zika Virus Infection/epidemiology ; Adult ; Zika Virus/immunology ; Female ; Male ; Interferon-gamma/metabolism ; Interferon-gamma/immunology ; Brazil/epidemiology ; CD8-Positive T-Lymphocytes/immunology ; CD4-Positive T-Lymphocytes/immunology ; Middle Aged ; Young Adult ; Epidemics ; Lymphocyte Activation/immunology ; T-Lymphocytes/immunology
Chemical Substances	Interferon-gamma (82115-62-6)
Language	English
Publishing date	2024-05-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0302684
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Article ; Online: Single-cell mass cytometry on peripheral cells in Myasthenia Gravis identifies dysregulation of innate immune cells.

Verdier, Julien / Fayet, Odessa-Maud / Hemery, Edouard / Truffault, Frédérique / Pinzón, Natalia / Demeret, Sophie / Behin, Anthony / Fadel, Elie / Guihaire, Julien / Corneau, Aurélien / Blanc, Catherine / Berrih-Aknin, Sonia / Le Panse, Rozen

Frontiers in immunology

2023 Volume 14, Page(s) 1083218

Abstract: Myasthenia Gravis (MG) is a neurological autoimmune disease characterized by disabling muscle weaknesses due to anti-acetylcholine receptor (AChR) autoantibodies. To gain insight into immune dysregulation underlying early-onset ... ...

Abstract	Myasthenia Gravis (MG) is a neurological autoimmune disease characterized by disabling muscle weaknesses due to anti-acetylcholine receptor (AChR) autoantibodies. To gain insight into immune dysregulation underlying early-onset AChR
MeSH term(s)	Humans ; Immunity, Innate ; Lymphocytes ; Myasthenia Gravis ; Receptors, Cholinergic ; Autoantibodies ; Thymus Neoplasms ; Nervous System Diseases
Chemical Substances	Receptors, Cholinergic ; Autoantibodies
Language	English
Publishing date	2023-01-30
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2023.1083218
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Article ; Online: Mass cytometry reveals atypical immune profile notably impaired maturation of memory CD4 T with Gb3-related CD27 expression in CD4 T cells in Fabry disease.

Journal of inherited metabolic disease

2024

Abstract: Fabry disease (FD) is an X-linked disease characterized by an accumulation of glycosphingolipids, notably of globotriaosylceramide (Gb3) and globotriaosylsphingosine (lysoGb3) leading to renal failure, cardiomyopathy, and cerebral strokes. Inflammatory ... ...

Abstract	Fabry disease (FD) is an X-linked disease characterized by an accumulation of glycosphingolipids, notably of globotriaosylceramide (Gb3) and globotriaosylsphingosine (lysoGb3) leading to renal failure, cardiomyopathy, and cerebral strokes. Inflammatory processes are involved in the pathophysiology. We investigated the immunological phenotype of peripheral blood mononuclear cells in Fabry patients depending on the clinical phenotype, treatment, Gb3, and lysoGb3 levels and the presence of anti-drug antibodies (ADA). Leucocytes from 41 male patients and 20 controls were analyzed with mass cytometry using both unsupervised and supervised algorithms. FD patients had an increased expression of CD27 and CD28 in memory CD45- and CD45 + CCR7-CD4 T cells (respectively p < 0.014 and p < 0.02). Percentage of CD45RA-CCR7-CD27 + CD28+ cells in CD4 T cells was correlated with plasma lysoGb3 (r = 0.60; p = 0.0036) and phenotype (p < 0.003). The correlation between Gb3 and CD27 in CD4 T cells almost reached significance (r = 0.33; p = 0.058). There was no immune profile associated with the presence of ADA. Treatment with agalsidase beta was associated with an increased proportion of Natural Killer cells. These findings provide valuable insights for understanding FD, linking Gb3 accumulation to inflammation, and proposing new prognostic biomarkers.
Language	English
Publishing date	2024-04-16
Publishing country	United States
Document type	Journal Article
ZDB-ID	438341-2
ISSN	1573-2665 ; 0141-8955
ISSN (online)	1573-2665
ISSN	0141-8955
DOI	10.1002/jimd.12727
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Article ; Online: Immune landscape after allo-HSCT: TIGIT- and CD161-expressing CD4 T cells are associated with subsequent leukemia relapse.

Gournay, Viviane / Vallet, Nicolas / Peux, Vivien / Vera, Kristi / Bordenave, Jennifer / Lambert, Marion / Corneau, Aurélien / Michonneau, David / Peffault de Latour, Régis / Caillat-Zucman, Sophie / Socié, Gérard / Chevalier, Mathieu F

Blood

2022 Volume 140, Issue 11, Page(s) 1305–1321

Abstract: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the most effective treatment for selected patients with acute myeloid leukemia (AML) and relies on a "graft-versus-leukemia" effect (GVL) where donor T lymphocytes mediate control of ... ...

Abstract	Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the most effective treatment for selected patients with acute myeloid leukemia (AML) and relies on a "graft-versus-leukemia" effect (GVL) where donor T lymphocytes mediate control of malignant cell growth. However, relapse remains the major cause of death after allo-HSCT. In various malignancies, several immunoregulatory mechanisms have been shown to restrain antitumor immunity, including ligand-mediated engagement of inhibitory receptors (IRs) on effector cells, and induction of immunosuppressive cell subsets, such as regulatory T cells (Tregs) or myeloid-derived suppressor cells (MDSCs). Relapse after HSCT remains a major therapeutic challenge, but immunoregulatory mechanisms involved in restraining the GVL effect must be better deciphered in humans. We used mass cytometry to comprehensively characterize circulating leukocytes in 2 cohorts of patients after allo-HSCT. We first longitudinally assessed various immunoregulatory parameters highlighting specific trends, such as opposite dynamics between MDSCs and Tregs. More generally, the immune landscape was stable from months 3 to 6, whereas many variations occurred from months 6 to 12 after HSCT. Comparison with healthy individuals revealed that profound alterations in the immune equilibrium persisted 1 year after HSCT. Importantly, we found that high levels of TIGIT and CD161 expression on CD4 T cells at month 3 after HSCT were distinct features significantly associated with subsequent AML relapse in a second cross-sectional cohort. Altogether, these data provide global insights into the reconstitution of the immunoregulatory landscape after HSCT and highlight non-canonical IRs associated with relapse, which could open the path to new prognostic tools or therapeutic targets to restore subverted anti-AML immunity.
MeSH term(s)	CD4-Positive T-Lymphocytes/pathology ; Cross-Sectional Studies ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myeloid, Acute/pathology ; Leukemia, Myeloid, Acute/therapy ; Ligands ; Receptors, Immunologic ; Recurrence ; Transplantation, Homologous
Chemical Substances	Ligands ; Receptors, Immunologic ; TIGIT protein, human
Language	English
Publishing date	2022-07-09
Publishing country	United States
Document type	Journal Article
ZDB-ID	80069-7
ISSN	1528-0020 ; 0006-4971
ISSN (online)	1528-0020
ISSN	0006-4971
DOI	10.1182/blood.2022015522
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Article ; Online: Analysis of cell surface and intranuclear markers on non-stimulated human PBMC using mass cytometry.

Dzangué-Tchoupou, Gaëlle / Corneau, Aurélien / Blanc, Catherine / Benveniste, Olivier / Allenbach, Yves

PloS one

2018 Volume 13, Issue 3, Page(s) e0194593

Abstract: Mass cytometry is a powerful tool that allows simultaneous analysis of more than 37 markers at the single cell level. Mass cytometry is of particular interest in the identification of a wide variety of cell phenotypes in autoimmune diseases. Moreover, ... ...

Abstract	Mass cytometry is a powerful tool that allows simultaneous analysis of more than 37 markers at the single cell level. Mass cytometry is of particular interest in the identification of a wide variety of cell phenotypes in autoimmune diseases. Moreover, cells can be labelled with palladium isotopes and pooled before staining (barcoding). Nevertheless, immunologists often face an important problem concerning the choice of markers to be included in a panel. This problem arises due to the incompatibility of different buffers used for the fixation and permeabilization of cells with various cell surface epitopes. In this study, we used a panel of 27 markers (19 surface markers and 8 intranuclear markers) to demonstrate disparities in the detection of cell surface antigens when comparing different buffers to stain unstimulated peripheral blood mononuclear cells. These disparities range from mild differences to very important differences in population frequencies depending on the buffers. Finally, we demonstrate the harmful effects of permeabilization prior to barcoding on the detection of some cell surface antigens. Here, we optimize a protocol that is suitable to use when targeting a large panel including both cell surface and intranuclear markers on unstimulated human peripheral blood mononuclear cells.
MeSH term(s)	Antigens, Surface/analysis ; Antigens, Surface/metabolism ; Biomarkers/analysis ; Cell Membrane/drug effects ; Cell Membrane/immunology ; Cell Membrane/metabolism ; Cell Membrane Permeability/drug effects ; Cell Nucleus/drug effects ; Cell Nucleus/immunology ; Cell Nucleus/metabolism ; Cell Separation/methods ; Epitopes/analysis ; Epitopes/metabolism ; Fixatives/adverse effects ; Flow Cytometry/methods ; Humans ; Isotopes/chemistry ; Leukocytes, Mononuclear ; Molecular Typing/methods ; Palladium/chemistry ; Staining and Labeling/methods ; Tissue Fixation/methods
Chemical Substances	Antigens, Surface ; Biomarkers ; Epitopes ; Fixatives ; Isotopes ; Palladium (5TWQ1V240M)
Language	English
Publishing date	2018
Publishing country	United States
Document type	Comparative Study ; Journal Article
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0194593
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Article ; Online: Que la lumière soit. Et si ce n’était plus seulement vrai ! - Évolution de la cytométrie : du conventionnel à la masse (en passant par le spectre).

Idziorek, Thierry / Cazareth, Julie / Blanc, Catherine / Jouy, Nathalie / Bourdely, Pierre / Corneau, Aurélien

Medecine sciences : M/S

2018 Volume 34, Issue 5, Page(s) 439–447

Abstract: The last decade has been an era of accelerated technological progress for flow cytometry. New technologies have been developed such as mass cytometry in which standard fluorochromes have been replaced by lanthanide-based non-radioactive metals and by ... ...

Title translation	Fiat Lux. May be no more true in cytometry! Go to mass and spectrum but still stay classic.
Abstract	The last decade has been an era of accelerated technological progress for flow cytometry. New technologies have been developed such as mass cytometry in which standard fluorochromes have been replaced by lanthanide-based non-radioactive metals and by spectral cytometry that measures the complete fluorescence spectrum. In this review, we schematically describe conventional, mass and spectral cytometry and present the plus and minus of each technology.
MeSH term(s)	Animals ; Flow Cytometry/instrumentation ; Flow Cytometry/methods ; Flow Cytometry/trends ; Fluorescence ; Fluorescent Dyes/chemistry ; Fluorescent Dyes/pharmacology ; Humans ; Light ; Mass Spectrometry/instrumentation ; Mass Spectrometry/methods ; Spectrum Analysis/instrumentation ; Spectrum Analysis/methods
Chemical Substances	Fluorescent Dyes
Language	French
Publishing date	2018-06-13
Publishing country	France
Document type	Journal Article ; Review
ZDB-ID	632733-3
ISSN	1958-5381 ; 0767-0974
ISSN (online)	1958-5381
ISSN	0767-0974
DOI	10.1051/medsci/20183405017
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Article ; Online: CD8+PD-L1+CXCR3+ polyfunctional T cell abundances are associated with survival in critical SARS-CoV-2-infected patients.

Adam, Lucille / Rosenbaum, Pierre / Quentric, Paul / Parizot, Christophe / Bonduelle, Olivia / Guillou, Noëlline / Corneau, Aurélien / Dorgham, Karim / Miyara, Makoto / Luyt, Charles-Edouard / Guihot, Amélie / Gorochov, Guy / Combadière, Christophe / Combadière, Behazine

JCI insight

2021 Volume 6, Issue 18

Abstract: The importance of the adaptive T cell response in the control and resolution of viral infection has been well established. However, the nature of T cell-mediated viral control mechanisms in life-threatening stages of COVID-19 has yet to be determined. ... ...

Abstract	The importance of the adaptive T cell response in the control and resolution of viral infection has been well established. However, the nature of T cell-mediated viral control mechanisms in life-threatening stages of COVID-19 has yet to be determined. The aim of the present study was to determine the function and phenotype of T cell populations associated with survival or death of patients with COVID-19 in intensive care as a result of phenotypic and functional profiling by mass cytometry. Increased frequencies of circulating, polyfunctional CD4+CXCR5+HLA-DR+ stem cell memory T cells (Tscms) and decreased proportions of granzyme B-expressing and perforin-expressing effector memory T cells were detected in recovered and deceased patients, respectively. The higher abundance of polyfunctional PD-L1+CXCR3+CD8+ effector T cells (Teffs), CXCR5+HLA-DR+ Tscms, and anti-nucleocapsid (anti-NC) cytokine-producing T cells permitted us to differentiate between recovered and deceased patients. The results from a principal component analysis show an imbalance in the T cell compartment that allowed for the separation of recovered and deceased patients. The paucity of circulating PD-L1+CXCR3+CD8+ Teffs and NC-specific CD8+ T cells accurately forecasts fatal disease outcome. This study provides insight into the nature of the T cell populations involved in the control of COVID-19 and therefore might impact T cell-based vaccine designs for this infectious disease.
MeSH term(s)	Adult ; B7-H1 Antigen/immunology ; CD4-Positive T-Lymphocytes/immunology ; CD8 Antigens/immunology ; CD8-Positive T-Lymphocytes/immunology ; COVID-19/immunology ; COVID-19/mortality ; COVID-19/pathology ; Epitopes, T-Lymphocyte/immunology ; Female ; France/epidemiology ; Humans ; Immunity, Cellular ; Immunologic Memory ; Lymphocyte Activation ; Male ; Receptors, CXCR3/immunology ; SARS-CoV-2 ; Survival Rate/trends
Chemical Substances	B7-H1 Antigen ; CD274 protein, human ; CD8 Antigens ; CXCR3 protein, human ; Epitopes, T-Lymphocyte ; Receptors, CXCR3
Language	English
Publishing date	2021-09-22
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2379-3708
ISSN (online)	2379-3708
DOI	10.1172/jci.insight.151571
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Article ; Online: VEXAS syndrome is characterized by inflammasome activation and monocyte dysregulation.

Kosmider, Olivier / Possémé, Céline / Templé, Marie / Corneau, Aurélien / Carbone, Francesco / Duroyon, Eugénie / Breillat, Paul / Chirayath, Twinu-Wilson / Oules, Bénédicte / Sohier, Pierre / Luka, Marine / Gobeaux, Camille / Lazaro, Estibaliz / Outh, Roderau / Le Guenno, Guillaume / Lifermann, François / Berleur, Marie / Le Mene, Melchior / Friedrich, Chloé /

Lenormand, Cédric / Weitten, Thierry / Guillotin, Vivien / Burroni, Barbara / Boussier, Jeremy / Willems, Lise / Aractingi, Selim / Dionet, Léa / Tharaux, Pierre-Louis / Vergier, Béatrice / Raynaud, Pierre / Ea, Hang-Korng / Ménager, Mickael / Duffy, Darragh / Terrier, Benjamin

Nature communications

2024 Volume 15, Issue 1, Page(s) 910

Abstract: Acquired mutations in the UBA1 gene were recently identified in patients with severe adult-onset auto-inflammatory syndrome called VEXAS (vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic). However, the precise physiological and clinical impact of ...

Abstract	Acquired mutations in the UBA1 gene were recently identified in patients with severe adult-onset auto-inflammatory syndrome called VEXAS (vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic). However, the precise physiological and clinical impact of these mutations remains poorly defined. Here we study a unique prospective cohort of VEXAS patients. We show that monocytes from VEXAS are quantitatively and qualitatively impaired and display features of exhaustion with aberrant expression of chemokine receptors. In peripheral blood from VEXAS patients, we identify an increase in circulating levels of many proinflammatory cytokines, including IL-1β and IL-18 which reflect inflammasome activation and markers of myeloid cells dysregulation. Gene expression analysis of whole blood confirms these findings and also reveals a significant enrichment of TNF-α and NFκB signaling pathways that can mediate cell death and inflammation. This study suggests that the control of the nflammasome activation and inflammatory cell death could be therapeutic targets in VEXAS syndrome.
MeSH term(s)	Adult ; Humans ; Monocytes ; Inflammasomes/genetics ; Prospective Studies ; Myeloid Cells ; Mutation ; Myelodysplastic Syndromes ; Skin Diseases, Genetic
Chemical Substances	Inflammasomes
Language	English
Publishing date	2024-01-30
Publishing country	England
Document type	Journal Article
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-024-44811-4
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