scholarly journals Proteolytic Processing of Stat6 Signaling in Mast Cells as a Negative Regulatory Mechanism

2002 ◽  
Vol 196 (1) ◽  
pp. 27-38 ◽  
Author(s):  
Kotaro Suzuki ◽  
Hiroshi Nakajima ◽  
Shin-ichiro Kagami ◽  
Akira Suto ◽  
Kei Ikeda ◽  
...  
Keyword(s):  
Mast Cells ◽  
Regulatory Mechanism ◽  
Allergic Diseases ◽  
Proteolytic Processing ◽  
Dominant Negative ◽  
Negative Regulator ◽  
Nuclear Accumulation ◽  
Transactivation Domain ◽  
Negative Regulatory Mechanism

Accumulating evidence has shown the importance of Stat6-mediated signaling in allergic diseases. In this study, we show a novel regulatory mechanism of Stat6-mediated signaling in mast cells. When Stat6 is activated by interleukin (IL)-4 and translocated to the nucleus, Stat6 is cleaved by a nucleus-associated protease in mast cells. The cleaved 65-kD Stat6 lacks the COOH-terminal transactivation domain and functions as a dominant-negative molecule to Stat6-mediated transcription. The retrovirus-mediated expression of cleavage-resistant Stat6 mutants prolongs the nuclear accumulation of Stat6 upon IL-4 stimulation and enhances IL-4–induced gene expression and growth inhibition in mast cells. These results indicate that the proteolytic processing of Stat6 functions as a lineage-specific negative regulator of Stat6-dependent signaling in mast cells, and thus suggest that it may account for the limited role of Stat6 in IL-4 signaling in mast cells.

Proteomic Analysis of the Role of STAT5A in the Apoptosis of Human Pre-B Cells : Evidence for a Connection to Daxx and the Oxidative Stress

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 3858-3858
Author(s):  
Eric Cholez ◽  
Veronique Debuysscher ◽  
Roland Charlionet ◽  
Francois Tron ◽  
Fabrice Gouilleux ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
B Cell ◽  
Cellular Proliferation ◽  
Dominant Negative ◽  
Proliferation Rate ◽  
Negative Regulator ◽  
Glutathione Synthetase ◽  
Transactivation Domain ◽  
Proteomic Approach

Abstract It is well established that STAT5A and 5B transcription factors (TF) play a major role in hematopoiseis and oncogenesis. In particular, there is a body of evidence suggesting that they might be involved in B lymphocyte survival, proliferation and development as well as in B cell neoplastic transformation. To investigate the role of STAT5A in human precursor B cell survival, we stably transfected a dominant negative form of STAT5A (DN-STAT5A, deleted in its transactivation domain) in the human leukemic Nalm6 pre-B cell line. All clones expressing DN-STAT5A exhibited a lower proliferation rate associated with a higher spontaneous apoptosis and that was massively enhanced upon IL7 stimulation. They also were more sensitive to FAS (CD95)- and at a lesser extent to etoposide-induced cell death than cells transfected with the empty vector (Nalm6neo), suggesting a hitherto unknown link between STAT5A and apoptosis/survival pathways. There was no evidence for changes in the levels of expression of Bcl2, BclxL, Bad or –c-Myc in DN-STAT5A expressing cells whereas Bax and Bim expression was sharply up-regulated in these cells. In order to identify the proteins involved in the connection between Fas and STAT5A, we used a proteomic approach with differential analysis of cells expressing (Nalm6Δ5A749) or not (Nalm6neo) the DN-STAT5A. Statistical analysis of bidimensional (2D) gels and use of MALDI-TOF technique enabled us to identify 7 proteins down-regulated in the Nalm6Δ5A749 cells, including members of heat shock proteins such as hsp27 and hsp70, as well as proteins implied in the control of oxidative stress like glutathione synthetase and transaldolase. By contrast, whereas 6 proteins were shown to be up-regulated, including prohibitin which is a negative regulator of cellular proliferation. Nalm6Δ5A749 cells exhibited enhanced levels of reactive oxygen species in keeping with the idea that oxidative stress might be involved in the increased sensitivity of these cells to apoptotic signals. Although expression of all above mentioned molecules did not change upon Fas stimulation, addition of gluthatione (10mM) resulted in a complete inhibition of Fas-mediated apoptosis in Nalm6Δ5A749 cells. Whether or not prohibitin might be involved in the decreased proliferation rate in DN-STAT5A expressing cells needs further investigation. As Hsp27 has been previously shown to have the potential to sequestrate Daxx, a molecule that is involved in Fas receptor signaling, we investigated the subcellular location of this molecule in Nalm6Δ5A749 cells. We showed that Daxx was essentially expressed in the cytoplasm of these cells whereas it was mainly located in the nucleus in Nalm6neo cells. Our results bring evidence for a role of STAT5A in pre-B cell growth and survival and in particular for a connection between this TF and Fas signaling machinery that might involve Hsp27 and Daxx. They also point out to a so far undescribed link between STAT5 and oxidative stress.

Gtpase Immunity-Associated Protein Family Member 4 Contributes to the Enhanced Expansion of HSPCs in RUNX1 Deficient Mice

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4344-4344
Author(s):  
Amanda Scholl ◽  
Kentson Lam ◽  
Alex Muselman ◽  
Tingdong Tang ◽  
Shinobu Matsuura ◽  
...  
Keyword(s):  
Family Member ◽  
Conflicts Of Interest ◽  
Protein Family ◽  
Dominant Negative ◽  
Negative Regulator ◽  
Wild Type ◽  
Double Knockout ◽  
Hematopoietic Stem ◽  
Myeloid Progenitor

Abstract RUNX1 is the transcription factor described as the master regulator of hematopoiesis. Due to its central role during blood development, numerous RUNX1 mutations have been reported in hematologic abnormalities. Mice null for Runx1 die during embryogenesis, lacking definitive HSCs. Conditional Runx1Δ/Δ mice are viable, but exhibit a variety of blood abnormalities. The most salient defect in these Runx1Δ/Δ mice is expansion of the hematopoietic stem and progenitor cell (HSPC) population, measured as an increase in number of lineage negative, Sca1 positive, cKit positive (LSK) cells. A shortened form of RUNX1 (RUNX1SF) lacking the C-terminal and part of the N-terminal domain (41-214) acts as a dominant negative regulator of RUNX1 and hence also models RUNX1 loss-of-function. A differential gene expression analysis of HSPCs derived from Runx1Δ/Δ compared to wild type mice uncovered GTPase immunity-associated protein family member 4 (GIMAP4) as one of the genes most highly upregulated. Previous studies have focused almost exclusively on the role of GIMAP4 as a pro-apoptotic protein during T-cell development. This study illuminates a novel non-apoptotic role of GIMAP4 in a formerly unstudied HSPC context. Runx1Δ/Δ mice were crossed with Gimap4-/- mice to generate a double knockout (dKO) mouse line. These dKO mice exhibited attenuated HSPC proliferation in comparison to Runx1Δ/Δ mice, suggesting that GIMAP4 functions in this HSPC expansion phenotype. BMT experiments using lethally irradiated C57 mice and RUNX1SF transduced wild type versus Gimap4-/-bone marrow confirmed this result. GIMAP4 also worked independently and coordinately with RUNX1 to influence individual progenitor populations. Common lymphoid progenitors (CLP) were affected only by GIMAP4. Gimap4-/- mice exhibited an expansion of the CLP population, consistent with its pro-apoptotic role in lymphoid populations. Conversely, both RUNX1 and GIMAP4 coordinately exerted an effect on myeloid progenitor populations. Runx1Δ/Δ mice harbored expanded granulocyte-macrophage progenitor (GMP) and common myeloid progenitor (CMP) populations. This expansion was not observed when GIMAP4 was also ablated. This suggests a pro-proliferative role of GIMAP4 specifically in myeloid populations. These opposing roles of GIMAP4 in lymphoid versus myeloid cells suggest a more contextual, cell-specific role of this GTPase protein. Ultimately, this study provides insight into how RUNX1 and GIMAP4 may coordinate to maintain HSPC homeostasis. Disclosures No relevant conflicts of interest to declare.

scholarly journals The Role of Interleukin-37 in the Pathogenesis of Allergic Diseases

Acta Naturae ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 54-64
Author(s):  
I. P. Shilovskiy ◽  
M. E. Dyneva ◽  
O. M. Kurbacheva ◽  
D. A. Kudlay ◽  
M. R. Khaitov
Keyword(s):  
Allergic Rhinitis ◽  
Atopic Dermatitis ◽  
Bronchial Asthma ◽  
Interleukin 1 ◽  
Allergic Diseases ◽  
Biological Properties ◽  
Negative Regulator ◽  
Cellular Mechanisms ◽  
Wide Range

Cytokines of the interleukin-1 (IL-1) family play an important role in the realization of the protective functions of innate immunity and are the key mediators involved in the pathogenesis of a wide range of diseases, including various manifestations of allergy. The IL-1 family includes more than 11 members. However, the functions of many of them remain to be elucidated. Recently, new members of the IL-1 family have been discovered. In 2000, several independent research groups reported the discovery of a new interleukin of this family, which was named IL-37, or IL-1F7 (according to the new nomenclature). IL-37 was assigned to the IL-1 family based on its structural similarity with other members of this family. The study of its biological properties showed that its activity changes in inflammatory diseases, such as rheumatoid arthritis, psoriasis, as well as allergic diseases (allergic rhinitis, bronchial asthma, and atopic dermatitis). However, unlike most members of the IL-1 family, IL-37 acts as a negative regulator of inflammation. Activation of IL-37 suppresses inflammation, resulting in the suppression of inflammatory cytokines and chemokines, which in turn prevents infiltration of pro-inflammatory cells, mainly eosinophils and neutrophils. The exact molecular and cellular mechanisms of the anti-inflammatory effect of IL-37 in the development of allergic diseases (AD) have not been fully studied. This review summarizes and analyzes the accumulated experimental data on the role of IL-37 in the pathogenesis of AD, such as allergic rhinitis, bronchial asthma, and atopic dermatitis.

Historical anecdotes and breakthroughs of histamine: from discovery to date

2020 ◽  
Vol 20 ◽  
Author(s):  
Ioannis Alexandros Charitos ◽  
Francesca Castellaneta ◽  
Luigi Santacroce ◽  
Lucrezia Bottalico
Keyword(s):  
Mast Cells ◽  
Treatment Guidelines ◽  
Allergic Diseases ◽  
History Of ◽  
Museum Archives ◽  
Ancient Times ◽  
New Research ◽  
Preclinical And Clinical Studies ◽  
Historical References

Aim: Investigating about the history of allergies and discovery of the histamine’s role in the immune response through historical references, starting with ancient anecdotes, analysing the first immunization attempts on animals to understand its importance as the anaphylaxis mediator. Moreover, we shortly resume the most recent discoveries on mast cell role in allergic diseases throughout the latest updates on its antibody-independent receptors. Methods: Publications, including reviews, treatment guidelines, historical and medical books, on the topic of interest were found on Medline, PubMed, Web of Knowledge, Web of Science, Google Scholar, Elsevier’s (EMBASE.comvarious internet museum archives. Texts from the National Library of Greece (Stavros Niarchos Foundation), from the School of Health Sciences of the National and Kapodistrian University of Athens (Greece). We selected key articles which could provide an historical and scientific insight into histamine molecule and its mechanism of action’s discovery starting with Egyptian, Greek and Chinese antiquity to end with the more recent pharmacological and molecular discoveries. Results: Allergic diseases were described by medicine since ancient times, without exactly understanding physio-pathologic mechanisms of immuno-mediated reactions and of their most important biochemical mediator, histamine. Researches on histamine and allergic mechanisms started at the beginning of the 20th century with the first experimental observations on animals of anaphylactic reactions. Histamine was then identified as their major mediator of many allergic diseases and anaphylaxis, but also of several physiologic body’s functions, and its four receptors were characterized. Modern researches focus their attention on the fundamental role of the antibody-independent receptors of mast cells in allergic mechanisms, such as MRGPRX2, ADGRE2 and IL-33 receptor. Conclusion: New research should investigate how to modulate immunity cells activity in order to better investigate possible multi-target therapies for host’s benefits in preclinical and clinical studies on allergic diseases in which mast cells play a major role.

OTT-MKL1 and MKL1 Inhibit Wnt Signaling.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2250-2250
Author(s):  
Stephanie Halene ◽  
Ee-chun Cheng ◽  
Vincent Schulz ◽  
David Tuck ◽  
Diane Krause
Keyword(s):  
Cell Death ◽  
Wnt Signaling ◽  
Transcriptional Activation ◽  
Wnt Pathway ◽  
Fusion Gene ◽  
Apoptotic Cell ◽  
Dominant Negative ◽  
Nuclear Accumulation ◽  
Transactivation Domain ◽  
Hel Cells

Abstract The OTT-MKL1 fusion gene product is generated as a result of t(1;22) in a subset of acute megakaryoblastic leukemia predominantly encountered in young children. Due to myelofibrosis and the age at presentation, patient samples are scarce. We generated Human Erythroid Leukemia (HEL) cell derived cell lines with tet-inducible OTT, MKL1 and OTT-MKL1 to further elucidate the function of the respective proteins. HEL cells can be induced to differentiate down the megakaryocyte lineage by TPA. Induction with doxycycline resulted in transcription and translation of the respective genes within hours. While overexpression of MKL1 led to enhancement of megakaryocytic differentiation, both OTT and OTT-MKL1 overexpression led to cell death over the course of several days by apoptosis as evident by staining for Annexin V and morphology. The apoptotic cell death was greatly enhanced by concomitant induction of differentiation by TPA. We performed microarray analysis comparing uninduced and 8-hour tet-induced samples in the presence or absence of TPA. While overexpression of OTT had only a minimal effect on the transcriptome of the HEL cells, both MKL1 and OTT-MKL1 significantly affected the gene expression of many genes. Using a false discovery rate cut-off of p < 0.05, and assessing only those genes whose expression changed by greater than 2-fold, OTT-MKL1 and MKL1 induced the upregulation of 157 and 168 genes, respectively, and the downregulation of 56 and 62 genes, respectively. Only 20 genes were upregulated by both OTT-MKL1 and MKL1, and 12 genes were downregulated greater than 2-fold by both OTT-MKL1 and MKL1. GeneGo analysis comparing OTT-MKL1 over-expressing versus non-expressing cells revealed over-representation of the Wnt pathway. Among the differentially expressed genes implicated in the Wnt pathway were Frat1 and Frat2, which have been shown to inhibit GSK3β and lead to β-catenin nuclear accumulation, and thus stimulation of the Wnt pathway. At the same time, inhibitory NLK was upregulated and several down-stream targets of the Wnt pathway were downregulated. Spenito, the homolog of OTT in Drosophila, is known to have promoter-specific activating and inhibiting effects on Wnt target genes. We thus performed reporter assays to study the effects of OTT, MKL1 and OTT-MKL1 on the Wnt pathway. Using the TOP-/FOP-FLASH reporter system in 293T cells, there was a dose-dependent inhibition of β-catenin-mediated activation of the Tcf/Lef binding site promoter by MKL1 and OTT-MKL1. Full length OTT showed a minimal stimulatory effect only at low doses., while N-terminal OTT, lacking the SPOC domain and a dominant negative form of MKL1 lacking the transactivation domain each enhanced β-catenin induced Tcf/Lef mediated transcriptional activation. Studies to define the domains of OTT and MKL1 and the underlying mechanisms in hematopoietic cells are underway. These results suggest that MKL1 and OTT-MKL1 inhibit canonical Wnt signaling by inhibiting β-catenin induced transcription.

scholarly journals Mast cells in collagen diseases

2019 ◽  
Vol 3 (2) ◽  
pp. 69
Author(s):  
Naoya Mikita ◽  
Yutaka Inaba ◽  
Takashi Yoshimasu ◽  
Nobuo Kanazawa ◽  
Fukumi Furukawa
Keyword(s):  
Rheumatoid Arthritis ◽  
Multiple Sclerosis ◽  
Mast Cells ◽  
Immune Cells ◽  
Lupus Erythematosus ◽  
Allergic Diseases ◽  
Local Inflammation ◽  
Systemic Lupus ◽  
Cytokines And Chemokines

Mast cells are involved in many immune reactions and diseases through 1) the expressions of several receptors, 2) productions of various mediators such as histamine, cytokines, and chemokines, 3) direct interactions with immune cells. Besides allergic diseases, mast cells have been also assumed to be involved in autoimmune diseases such as bullous pemphigoid, rheumatoid arthritis, and multiple sclerosis. Moreover, several studies reported the involvement of mast cells in collagen disease. In this article, we review recent findings about the role of mast cells especially in systemic lupus erythematosus and systemic sclerosis. In these diseases, mast cells seem to be involved in local inflammation and tissue damage partially in the targeted organ rather than the development of autoimmunity including production of autoantibodies.

scholarly journals Mast cells in collagen diseases

2017 ◽  
Vol 1 (2) ◽  
pp. 75
Author(s):  
Naoya Mikita ◽  
Yutaka Inaba ◽  
Takashi Yoshimasu ◽  
Nobuo Kanazawa ◽  
Fukumi Furukawa
Keyword(s):  
Rheumatoid Arthritis ◽  
Multiple Sclerosis ◽  
Mast Cells ◽  
Immune Cells ◽  
Lupus Erythematosus ◽  
Allergic Diseases ◽  
Local Inflammation ◽  
Systemic Lupus ◽  
Cytokines And Chemokines

Mast cells are involved in many immune reactions and diseases through 1) the expressions of several receptors, 2) productions of various mediators such as histamine, cytokines, and chemokines, 3) direct interactions with immune cells. Besides allergic diseases, mast cells have been also assumed to be involved in autoimmune diseases such as bullous pemphigoid, rheumatoid arthritis, and multiple sclerosis. Moreover, several studies reported the involvement of mast cells in collagen disease. In this article, we review recent findings about the role of mast cells especially in systemic lupus erythematosus and systemic sclerosis. In these diseases, mast cells seem to be involved in local inflammation and tissue damage partially in the targeted organ rather than the development of autoimmunity including production of autoantibodies.

Identification of Oncostatin M as a Novel KIT D816V-Dependent Cytokine in Neoplastic Human Mast Cells.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 213-213
Author(s):  
Gregor Hoermann ◽  
Sabine Cerny-Reiterer ◽  
Andrea Perne ◽  
Miriam Klauser ◽  
Leonhard Muellauer ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mast Cell ◽  
Mast Cells ◽  
Dominant Negative ◽  
Oncostatin M ◽  
Mitogen Activated Protein Kinase ◽  
Affect Expression ◽  
Mitogen Activated Protein ◽  
Kit D816v

Abstract Abstract 213 Systemic mastocytosis (SM) is a neoplastic disease of mast cells (MC) and their bone marrow-derived progenitors. The clinical picture in SM is variable ranging from an indolent course to highly aggressive variants with short survival time. The pathologic hallmark in SM is the multifocal dense infiltrate of MC in the bone marrow. Other typical features of SM include alterations of the bone marrow microenvironment such as increased angiogenesis and fibrosis. In a majority of patients, MC display the KIT mutation D816V which affects the activation loop at the entrance to the enzymatic pocket of the KIT kinase. As a consequence, KIT D816V exhibits constitutive tyrosine kinase activity and promotes cytokine-independent differentiation of MC. However, so far, little is known about KIT D816V-dependent expression of pathogenetically relevant molecules in neoplastic MC. Oncostatin M (OSM) is a pleiotropic cytokine of the interleukin-6 family which is produced mainly by activated T cells and monocytes. OSM has been shown to inhibit cell growth in cell lines derived from solid tumors but to stimulate proliferation of fibroblasts and endothelial cells. Recently, it has been reported that OSM produced by activated MC promotes growth of human dermal fibroblasts. Moreover, it has been suggested that OSM stimulates growth of murine bone marrow-derived mast cells in a mast cell/fibroblast coculture. However, expression of OSM in neoplastic MC or a potential pathogenetic role of OSM in SM have not been examined so far. The aim of the present study was to analyze expression of OSM in neoplastic human MC and to determine the role of KIT D816V in OSM expression. As assessed by immunohistochemistry performed on bone marrow sections of patients with SM, typical spindle-shaped neoplastic MC were found to express OSM. Serial section-staining confirmed that tryptase-positive MC co-express OSM. Expression of OSM was found in neoplastic MC in all patients investigated (n=15) and in all variants of SM (indolent SM as well as aggressive variants) with comparable staining intensities. Preincubation of anti-OSM antibody with a specific blocking peptide resulted in a negative stain. In Ba/F3 cells, doxycycline-inducible expression of KIT D816V led to a substantial upregulation of OSM mRNA and OSM protein, whereas expression of wild type KIT did not affect expression of OSM. In addition, the KIT D816V-positive HMC-1.2 mast cell line was found to express OSM at high levels, whereas the KIT D816V-negative HMC-1.1 subclone expressed only baseline levels of OSM. Correspondingly, the KIT D816V-targeting drug midostaurine (PKC412) decreased the expression of OSM in HMC-1.2 cells as well as in KIT D816V-expressing Ba/F3 cells in a dose-dependent manner. To investigate signaling pathways involved in KIT D816V-dependent expression of OSM, we applied pharmacologic inhibitors and dominant negative-acting signaling molecules. We found that KIT D816V-dependent expression of OSM is inhibited by the mitogen-activated protein-kinase/extracellular signal-regulated kinase (MEK) inhibitor, PD98059, but not by the phosphoinositide 3-kinase inhibitor, LY294002. Expression of dominant negative mutants of signal transducer and activator of transcription 5 (STAT5) did not affect expression of OSM in KIT D816V-expressing cells. In summary, our data identify OSM as a novel cytokine expressed in neoplastic MC in patients with SM and show that KIT D816V directly promotes expression of OSM through activation of the mitogen-activated protein-kinase pathway. OSM may be an important KIT D816V-dependent effector promoting angiogenesis and fibrogenesis/sclerosis in patients with SM. Disclosures: No relevant conflicts of interest to declare.

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