The lysosomal protease cathepsin D is efficiently sorted to and secreted from regulated secretory compartments in the rat basophilic/mast cell line RBL

2000 ◽  
Vol 113 (18) ◽  
pp. 3289-3298 ◽  
Author(s):  
A. Dragonetti ◽  
M. Baldassarre ◽  
R. Castino ◽  
M. Demoz ◽  
A. Luini ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Cell Line ◽  
Cathepsin D ◽  
Secretory Granules ◽  
Bioactive Molecules ◽  
Blue Staining ◽  
Lysosomal Protease ◽  
Mannose 6 Phosphate ◽  
Mast Cell Line

Basophils and mast cells contain a peculiar class of inflammatory granules that discharge their content upon antigen-mediated crosslinking of IgE-membrane receptors. The pathways for granule biogenesis and exocytosis in these cells are still largely obscure. In this study we employed the rat basophilic leukemia (RBL)/mast cell line to verify the hypothesis that inflammatory granules share common bioactive molecules and functional properties with lysosomes. We demonstrate that inflammatory granules, as identified by the monoclonal 5G10 antibody (which recognises an integral membrane protein) or by Toluidine Blue staining, have an intralumenal acidic pH, possess lysosomal enzymes and are accessible by fluid-phase and membrane endocytosis markers. In addition, we studied the targeting, subcellular localisation and regulated secretion of the lysosomal aspartic protease cathepsin D (CD) as affected by IgE receptor stimulation in order to obtain information on the pathways for granule biogenesis and exocytosis. Stimulation with DNP-BSA of specific IgE-primed RBL cells led to a prompt release of processed forms of CD, along with other mature lysosomal hydrolases. This release could be prevented by addition of EGTA, indicating that it was dependent on extracellular calcium influx. Antigen stimulation also induced exocytosis of immature CD forms accumulated by ammonium chloride, suggesting the existence of an intermediate station in the pathway for granule biogenesis still sensitive to regulated exocytosis. The targeting of molecules to secretory granules may occur via either a mannose-6-phosphate-dependent or mannose-6-phosphate-independent pathway. We conclude that endosomes and lysosomes in basophils/mast cells can act as regulated secretory granules or actually identify with them.

Dexamethasone suppresses gene expression and production of IL-13 by human mast cell line and lung mast cells

1998 ◽  
Vol 102 (1) ◽  
pp. 134-142 ◽  
Author(s):  
Toshiaki Fushimi ◽  
Hiroshi Okayama ◽  
Sanae Shimura ◽  
Hiroki Saitoh ◽  
Kunio Shirato
Keyword(s):  
Gene Expression ◽  
Mast Cell ◽  
Mast Cells ◽  
Cell Line ◽  
Human Mast Cell ◽  
Mast Cell Line

scholarly journals Stimulated release of histamine by a rat mast cell line is inhibited during mitosis.

1984 ◽  
Vol 98 (6) ◽  
pp. 2250-2254 ◽  
Author(s):  
T R Hesketh ◽  
M A Beaven ◽  
J Rogers ◽  
B Burke ◽  
G B Warren
Keyword(s):  
Mast Cell ◽  
Plasma Membrane ◽  
Histamine Release ◽  
Cell Line ◽  
Secretory Granules ◽  
Antigen Stimulation ◽  
Interphase Cells ◽  
Mast Cell Line ◽  
Stimulation Of ◽  
Mitotic Cells

Stimulated histamine release was depressed at least tenfold in mitotic 2H3 rat basophilic cells when compared with interphase cells even though both contained comparable amounts of histamine. Antigen stimulation of IgE-sensitized interphase cells initiated an influx of Ca2+ that preceded secretion of histamine and a similar Ca2+ influx occurred in stimulated mitotic cells. This strongly suggests that during mitosis there is a dramatic inhibition of one or more of the steps on the pathway leading from elevated intracellular Ca2+ to the fusion of secretory granules with the plasma membrane.

Comparative Cytokine Release from Human Monocytes, Monocyte-Derived Immature Mast Cells, and a Human Mast Cell Line (HMC-1)

1994 ◽  
Vol 103 (4) ◽  
pp. 504-508 ◽  
Author(s):  
Jürgen Grabbe ◽  
P.i.a. Welker ◽  
Annelie Möller ◽  
Edgar Dippel ◽  
Leonie K Ashman ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Cell Line ◽  
Human Mast Cell ◽  
Human Monocytes ◽  
Cytokine Release ◽  
Mast Cell Line

Signaling pathways activated in a unique mast cell line where interleukin-3 supports survival and stem cell factor is required for a proliferative response

Blood ◽  
1996 ◽  
Vol 87 (9) ◽  
pp. 3655-3668 ◽  
Author(s):  
AM O'Farrell ◽  
M Ichihara ◽  
AL Mui ◽  
A Miyajima
Keyword(s):  
Mast Cell ◽  
Stem Cell ◽  
Mast Cells ◽  
Cell Line ◽  
Stem Cell Factor ◽  
Mapk Pathway ◽  
Interleukin 3 ◽  
Murine Mast Cell ◽  
Mast Cell Line

Stem cell factor (SCF) and interleukin-3 (IL-3) both act on several target hematopoietic populations, including mast cells. We have isolated a unique murine mast cell line, B6M, that is phenotypically similar to immature mast cells. For B6M cells, IL-3 is a survival factor and alone does not stimulate proliferation. SCF can stimulate proliferation of B6M cells, and together IL-3 and SCF synergize to stimulate optimal proliferation and long-term growth. A sustained induction of c-myc is observed only in the presence of SCF (with or without IL-3). In B6M cells, both IL-3 and SCF stimulate phosphorylation of Shc and activation of the Ras, Raf-1, MAPK pathway. Interestingly, IL-3 plus SCF synergistically activate MAPK. IL-3, but not SCF, leads to activation of Jak 2 and Stat 5 and induces pim-1 expression. From these data, we suggest that the induction of pim-1 and c-myc is independently regulated. Furthermore, IL-3-stimulated activation of the Jak 2/Stat 5 pathway, induction of pim-1, and activation of the Ras/MAPK pathway are insufficient to mediate proliferation of B6M cells. The unusual IL-3 response of B6M cells provides a useful model to dissect signals required for IL-3-stimulated survival and proliferation.

scholarly journals Synaptotagmin II Negatively Regulates Ca2+-triggered Exocytosis of Lysosomes in Mast Cells

1999 ◽  
Vol 189 (10) ◽  
pp. 1649-1658 ◽  
Author(s):  
Dana Baram ◽  
Roberto Adachi ◽  
Ora Medalia ◽  
Michael Tuvim ◽  
Burton F. Dickey ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Cathepsin D ◽  
Cell Biology ◽  
Secretory Granules ◽  
Triggered Release ◽  
Lysosomal Fraction ◽  
Regulated Secretion ◽  
Processed Form

Synaptotagmins (Syts) I and II are believed to act as Ca2+ sensors in the control of neurotransmission. Here we demonstrate that mast cells express Syt II in their lysosomal fraction. We further show that activation of mast cells by either aggregation of FcεRI or by Ca2+ ionophores results in exocytosis of lysosomes, in addition to the well documented exocytosis of their secretory granules. Syt II directly regulates lysosomal exocytosis, whereby overexpression of Syt II inhibited Ca2+-triggered release of the lysosomal processed form of cathepsin D, whereas suppression of Syt II expression markedly potentiated this release. These findings provide evidence for a novel function of Syt II in negatively regulating Ca2+-triggered exocytosis of lysosomes, and suggest that Syt II–regulated secretion from lysosomes may play an important role in mast cell biology.

In vitro inhibitory effect of rupatadine on histamine and TNF-α release from dispersed canine skin mast cells and the human mast cell line HMC-1

2000 ◽  
Vol 49 (7) ◽  
pp. 355-360 ◽  
Author(s):  
M. Queralt ◽  
P. Brazís ◽  
M. Merlos ◽  
F. de Mora ◽  
A. Puigdemont
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Cell Line ◽  
Inhibitory Effect ◽  
Human Mast Cell ◽  
Tnf Α ◽  
Mast Cell Line

FOG-1 represses GATA-1-dependent FcϵRI β-chain transcription: transcriptional mechanism of mast-cell-specific gene expression in mice

Blood ◽  
2006 ◽  
Vol 108 (1) ◽  
pp. 262-269 ◽  
Author(s):  
Keiko Maeda ◽  
Chiharu Nishiyama ◽  
Tomoko Tokura ◽  
Hiroyasu Nakano ◽  
Shunsuke Kanada ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Cell Line ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Hematopoietic Progenitor Cell ◽  
Specific Gene ◽  
Mast Cell Line ◽  
High Affinity Ige Receptor ◽  
Β Chain

Cell-type-specific transcription of mouse high-affinity IgE receptor (FcϵRI) β-chain is positively regulated by the transcription factor GATA-1. Although GATA-1 is expressed in erythroid cells, megakaryocytes, and mast cells, the expression of mouse FcϵRI β-chain is restricted to mast cells. In the present study, we characterized the role of GATA-associated cofactor FOG-1 in the regulation of the FcϵRI β-chain promoter. The expression levels of FOG-1, GATA-1, and β-chain in each hematopoietic cell line were analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. FOG-1 expression was higher in the β-chain-negative hematopoietic progenitor cell line Ba/F3 than in the β-chain-positive mast cell line PT18. By contrast, GATA-1 expression was similar when comparing the 2 cell lines. A transient reporter assay demonstrated that the β-chain promoter functioned in PT18 but not in Ba/F3 and that the transcription activity of the β-chain promoter in PT18 was markedly suppressed by overexpression of FOG-1. Although the activity of the β-chain promoter, which was upregulated by coexpression of GATA-1, was significantly suppressed by coexpression of FOG-1 in the simian kidney CV-1 cells (β-chain-, GATA-1-, and FOG-1-), the transactivation of the β-chain promoter by the GATA-1 mutant V205G, which cannot bind FOG-1, was not affected by coexpression of FOG-1. Further, overexpression of FOG-1 in PT18 resulted in decreases in cell surface expression of FcϵRI and β-chain transcription. Finally, suppression of FOG-1 expression using an siRNA approach resulted in increased β-chain promoter activity in Ba/F3. These results suggest that FOG-1 expression level regulates the GATA-1-dependent FcϵRI β-chain promoter. (Blood. 2006;108:262-269)

Abstract 1164: Mast Cells Contribute to Pathogenesis in the Progression of Abdominal Aortic Aneuyrsm

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Toshihiro Tsuruda ◽  
Johji Kato ◽  
Kinta Hatakeyama ◽  
Kunihide Nakamura ◽  
Takuroh Imamura ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Cell Line ◽  
Secretory Granules ◽  
Histological Study ◽  
Human Mast Cell ◽  
Pharmacological Intervention ◽  
Aortic Tissue ◽  
Positive Staining ◽  
Abdominal Aortic

Abdominal aortic aneurysm (AAA) is histologically characterized by medial degeneration and various degrees of chronic adventitial inflammation, while the mechanisms for progression of aneurysm are poorly understood. In the present study, we carried out histological study of AAA tissues of patients (75±1 years, Mean±SEM), and interventional animal and cell culture experiments to investigate the role of mast cells in the pathogenesis of AAA. The number of mast cells determined by positive staining for tryptase, a major neutral protease of human mast cell secretory granules, was found to increase in the outer media or adventitia of human AAA, compared to those in the control aorta (2.1±1.1/mm 2 (n=57) vs. 0.6±0.4/mm 2 (n=22), p<0.01), showing a positive correlation (r=0.442, p<0.01) between the cell number and the AAA diameter. Western blot demonstrated that the protein expressions of tryptase, stem cell factor (a ligand for c-kit) and the phosphorylation of c-kit were significantly (p<0.01) increased in the AAA tissues, compared with the control aorta. In the animal experiments, diameter of abdominal aorta of the control (+/+) rats was increased up to 55% at 14 days following peri-aortic application of 0.4 mol/L calcium chloride, but only a 13% increase in the mast cell-deficient mutant Ws/Ws rats. The AAA formation was accompanied by accumulation of mast cells, T-lymphocytes, interferon-γ positive cells and by activated matrix metalloproteinases (MMP)-2 and -9, reduced elastin levels and augmented angiogenesis in the aortic tissue, but these changes were much less in the Ws/Ws rats than in the control (+/+) rats. The pharmacological intervention with the mast cell stabilizer tranilast (400 mg/kg/day) for 14 days significantly (p<0.01) attenuated the AAA development in the control (+/+) rats. Co-culturing the mast cell line HMC-1 with the monocyte/macrophage cell line U937 significantly (p<0.01) augmented MMP-9 activity. Furthermore, 100 and 300 μmol/L of tranilast significantly (p<0.01) decreased MMP-9 activity in those cells. Collectively, these data suggest that mast cells may play important roles in the progression of AAA.

scholarly journals Disrupted Lipid Raft Shuttling of FcεRI by n-3 Polyunsaturated Fatty Acid Is Associated With Ligation of G Protein-Coupled Receptor 120 (GPR120) in Human Mast Cell Line LAD2

2020 ◽  
Vol 7 ◽  
Author(s):  
Xiaofeng Wang ◽  
Ramses Ilarraza ◽  
Brian P. Tancowny ◽  
Syed Benazir Alam ◽  
Marianna Kulka
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Cell Line ◽  
G Protein ◽  
Lipid Raft ◽  
Human Mast Cell ◽  
G Protein Coupled Receptor ◽  
Cell Functions ◽  
Mast Cell Line ◽  
G Protein Coupled

n-3 polyunsaturated fatty acids (PUFA) influences a variety of disease conditions, such as hypertension, heart disease, diabetes, cancer and allergic diseases, by modulating membrane constitution, inhibiting production of proinflammatory eicosanoids and cytokines, and binding to cell surface and nuclear receptors. We have previously shown that n-3 PUFA inhibit mast cell functions by disrupting high affinity IgE receptor (FcεRI) lipid raft partitioning and subsequent suppression of FcεRI signaling in mouse bone marrow-derived mast cells. However, it is still largely unknown how n-3 PUFA modulate human mast cell function, which could be attributed to multiple mechanisms. Using a human mast cell line (LAD2), we have shown similar modulating effects of n-3 PUFA on FcεRI lipid raft shuttling, FcεRI signaling, and mediator release after cell activation through FcεRI. We have further shown that these effects are at least partially associated with ligation of G protein-coupled receptor 120 expressed on LAD2 cells. This observation has advanced our mechanistic knowledge of n-3 PUFA's effect on mast cells and demonstrated the interplay between n-3 PUFA, lipid rafts, FcεRI, and G protein-coupled receptor 120. Future research in this direction may present new targets for nutritional intervention and therapeutic agents.

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