Pharmacologic Regulation of Mediator Generation and Release from the Murine Bone Marrow Derived Mast Cell

1985 ◽  
Vol 77 (1-2) ◽  
pp. 121-125 ◽  
Author(s):  
Robert A. Lewis ◽  
Jean-Louis Robin ◽  
Frank Austen
Keyword(s):  
Bone Marrow ◽  
Mast Cell ◽  
Murine Bone Marrow ◽  
Pharmacologic Regulation

scholarly journals Strain difference of murine bone marrow-derived mast cell functions

2005 ◽  
Vol 78 (3) ◽  
pp. 605-611 ◽  
Author(s):  
Junko Noguchi ◽  
Etsushi Kuroda ◽  
Uki Yamashita
Keyword(s):  
Bone Marrow ◽  
Mast Cell ◽  
Strain Difference ◽  
Murine Bone Marrow ◽  
Cell Functions

The C-Terminal Zinc Finger of GATA2 Is Necessary for Normalization of Aberrant Myelomonocytic Differentiation Induced by Activating PTPN11 Mutations.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3177-3177
Author(s):  
Zhenyun Yang ◽  
Cara S. Voorhorst ◽  
Leila Ndong ◽  
Fuqin Yin ◽  
Takako Kondo ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Transcription Factor ◽  
Mast Cell ◽  
Monocytic Differentiation ◽  
Rt Pcr ◽  
Hematopoietic Progenitors ◽  
Murine Bone Marrow ◽  
Factor Expression ◽  
Transcription Factor Expression ◽  
In Cells

Abstract Juvenile myelomonocytic leukemia (JMML) is a myeloproliferative disorder characterized by overproduction of myelomonocytic cells. Activating mutations of PTPN11, which encodes the protein tyrosine phosphatase, Shp2, are found in 35% of JMML patients. Murine bone marrow low density mononuclear cells (LDMNCs) expressing activating Shp2 mutants preferentially undergo myelomonocytic differentiation despite being subjected to conditions that typically support only mast cell development. Evaluation of hematopoietic cell-specific transcription factor expression revealed that GATA2 expression, needed for mast cell differentiation, is dramatically reduced, while, surprisingly, PU.1 expression is unchanged in cells expressing activating Shp2 mutants. In addition to lineage-specific transcription factors such as PU.1, however, c-jun also promotes monocytic differentiation by functioning as a co-activator of PU.1. Thus, we hypothesized that activating Shp2 mutations (Shp2D61Y or Shp2 E76K) induce increased c-jun expression permitting, in collaboration with PU.1, excessive monocytic differentiation and reduced GATA2 expression in hematopoietic progenitors. As a corollary, we hypothesized that ectopic expression of GATA2, but not of GATA2 lacking the C-terminal zinc finger (GATA2del330–407), which is needed for disruption of the PU.1-c-jun interaction, would normalize aberrant myelomonocytic differentiation induced by activating Shp2 mutants. Consistent with our hypothesis, quantitative RT-PCR studies revealed 5-fold higher c-jun levels in cells expressing Shp2D61Y or Shp2E76K compared to cells expressing WT Shp2. We next utilized retroviral co-transduction of murine bone marrow LDMNCs to generate six groups: pMIEG3-Shp2WT plus pCD4 (empty vector); pMIEG3-Shp2WT plus pCD4-GATA2; pMIEG3-Shp2WT plus pCD4-GATA2del330-407; pMIEG3-Shp2E76K plus pCD4; pMIEG3-Shp2E76K plus pCD4-GATA2; and pMIEG3-Shp-2E76K plus pCD4-GATA2del330-407. Transduced cells were stained with anti-human CD4 conjugated to allophycocyanin (APC), sorted for EGFP+APC+ cells, and plated into progenitor assays. Colonies were scored for colony forming unit (CFU)-granulocyte-macrophage (GM), monocyte (M), granulocyte (G), and granulocyte-erythroid-monocyte-megakaryocyte (GEMM). As predicted, cells co-transduced with activating Shp2E76K plus pCD4 produced significantly more CFU-M than cells co-transduced with WT Shp2 plus pCD4. Upon co-transduction with GATA2, the number of CFU-M generated from Shp2E76K-expressing cells was significantly reduced and was similar to that observed in cells expressing WT Shp2. In contrast, co-transduction of GATA2del330-407 failed to normalize the number of CFU-M produced by Shp2E76K-expressing cells. Quantitative RT-PCR verified ectopic GATA2 and GATA2del330-407 expression in the co-transduced cells. These findings demonstrate that restoration of GATA2 expression normalizes the propensity toward monocytic differentiation induced by Shp2E76K. The lack of correction conferred by GATA2del330-407 in combination with the observed increased c-jun expression support a model in which GATA2 and c-jun compete for binding to PU.1 to direct cell differentiation decisions in hematopoietic progenitors bearing activating Shp2 mutants. Collectively, these findings imply that normalization of transcription factor expression may provide a novel approach to differentiation-mediated therapy in JMML.

A self-renewing, bipotential erythroid/mast cell progenitor in continuous cultures of normal murine bone marrow

1985 ◽  
Vol 125 (1) ◽  
pp. 10-18 ◽  
Author(s):  
Fran�oise Wendling ◽  
Mona Shreeve ◽  
David McLeod ◽  
Arthur Axelrad
Keyword(s):  
Bone Marrow ◽  
Mast Cell ◽  
Continuous Cultures ◽  
Murine Bone Marrow ◽  
Mast Cell Progenitor

scholarly journals Dynamics of Plasma and Granule Membrane in Murine Bone Marrow-Derived Mast Cells after Re-stimulation

2015 ◽  
Vol 8 (1) ◽  
pp. 14-22
Author(s):  
Masahiro Kaneko ◽  
Arisa Yamada
Keyword(s):  
Bone Marrow ◽  
Mast Cell ◽  
Plasma Membrane ◽  
Mast Cells ◽  
Morphological Changes ◽  
Membrane Dynamics ◽  
Hematopoietic Stem ◽  
Murine Bone Marrow ◽  
Cell Functions ◽  
Granule Membrane

Mast cells are derived from hematopoietic stem cells and play important roles in allergic responses. Mast cells are long-lived compared with other granular cell types. Since the response of the individual mast cell after FcεRI-induced degranulation is unclear, the aim of this study was to analyze morphological changes in individual mast cells after restimulation. To observe plasma and granule membrane dynamics, AcGFP-actb (β-actin) and DsRed-monomer (DRM)- CD63 fusion constructs were introduced into bone marrow-derived mast cells (BMMCs). Furthermore, AcGFP-CD63 and DRM-Cma1 (mMCP-5) were introduced into BMMCs. Re-stimulation resulted in increased β-hexosaminidase release and cytokine mRNA expression similar to those observed during initial stimulation. Moreover, expression of FcεRI on BMMCs 24 h after initial stimulation was similar to that measured before initial stimulation. Changes in morphology of the plasma membrane and colocalization of granules and plasma membrane were observed after initial stimulation. BMMCs returned to normal 120 min after the initial stimulation. These phenomena were also observed in BMMCs after re-stimulation. BMMC chymase content decreased 20 min after stimulation but returned to near normal 24 h after stimulation. These findings suggest that mast cell functions can be maintained and that these cells can be repeatedly degranulated after FcεRI-mediated stimulation.

scholarly journals Identification of eosinophil lineage–committed progenitors in the murine bone marrow

2005 ◽  
Vol 201 (12) ◽  
pp. 1891-1897 ◽  
Author(s):  
Hiromi Iwasaki ◽  
Shin-ichi Mizuno ◽  
Robin Mayfield ◽  
Hirokazu Shigematsu ◽  
Yojiro Arinobu ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mast Cell ◽  
Bone Marrow Cells ◽  
Cell Level ◽  
Short Term ◽  
Monocyte Differentiation ◽  
Murine Bone Marrow ◽  
Cellular Target ◽  
Short Term Culture

Eosinophil lineage–committed progenitors (EoPs) are phenotypically isolatable in the steady-state murine bone marrow. Purified granulocyte/monocyte progenitors (GMPs) gave rise to eosinophils as well as neutrophils and monocytes at the single cell level. Within the short-term culture of GMPs, the eosinophil potential was found exclusively in cells activating the transgenic reporter for GATA-1, a transcription factor capable of instructing eosinophil lineage commitment. These GATA-1–activating cells possessed an IL-5Rα+CD34+c-Kitlo phenotype. Normal bone marrow cells also contained IL-5Rα+CD34+c-Kitlo EoPs that gave rise exclusively to eosinophils. EoPs significantly increased in number in response to helminth infection, suggesting that the EoP stage is physiologically involved in eosinophil production in vivo. EoPs expressed eosinophil-related genes, such as the eosinophil peroxidase and the major basic protein, but did not express basophil/mast cell–related mast cell proteases. The enforced retroviral expression of IL-5Rα in GMPs did not enhance the frequency of eosinophil lineage read-outs, whereas IL-5Rα+ GMPs displayed normal neutrophil/monocyte differentiation in the presence of IL-5 alone. Thus, IL-5Rα might be expressed specifically at the EoP stage as a result of commitment into the eosinophil lineage. The newly identified EoPs could be the cellular target in the treatment of a variety of disorders mediated by eosinophils.

scholarly journals Increased Growth Promoting But Not Mast Cell Degranulation Potential of a Covalent Dimer of c-Kit Ligand

Blood ◽  
1997 ◽  
Vol 90 (10) ◽  
pp. 3874-3883 ◽  
Author(s):  
Karl H. Nocka ◽  
Beth A. Levine ◽  
Jone-Lone Ko ◽  
Peter M. Burch ◽  
Bryan E. Landgraf ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mast Cell ◽  
Mast Cells ◽  
Cell Activation ◽  
Mast Cell Activation ◽  
Native Form ◽  
Murine Bone Marrow ◽  
Kit Ligand ◽  
Growth Promoting

Abstract The native form of soluble c-kit ligand (KL) is a noncovalent dimer. We have isolated a soluble, disulfide-linked dimer of murine KL (KL-CD) by expressing KL in Escherichia coli and refolding the denatured protein under conditions that promote the formation of both noncovalent dimers (KL-NC) and KL-CD. KL-CD exhibits a 10- to 15-fold increase in the ability to stimulate the growth of both the human megakaryocytic cell line MO7e and murine bone marrow-derived mast cells relative to KL-NC. Colony-forming assays of murine bone marrow progenitor cells also reflected this increased potency. However, KL-CD and KL-NC are equally able to prime mast cells for enhanced IgE-dependent degranulation in vitro and activate mast cells in vivo. Improving the growth-promoting activity of KL without changing its mast cell activation potential suggests that KL-CD or a related molecule could be administered in the clinic at doses that stimulate hematopoietic recovery while avoiding significant mast cell activation.

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