Identification of novel mast cell genes by serial analysis of gene expression in cord blood-derived mast cells

Mast cells are critical for allergic and inflammatory responses in which the peptide substance P (SP) and the cytokine IL-33 are involved. SP (0.01–1 μM) administered together with IL-33 (30 ng/mL) to human cultured LAD2 mast cells stimulates a marked increase (P< 0.0001) in secretion of the proinflammatory cytokine IL-1β. Preincubation of LAD2 (30 min) with the SP receptor (NK-1) antagonists L-733,060 (10 μM) or CP-96345 (10 µM) inhibits (P< 0.001) secretion of IL-1β stimulated by either SP (1 μM) or SP together with IL-33 (30 ng/mL). Surprisingly, secretion of IL-1β stimulated by IL-33 is inhibited (P< 0.001) by each NK-1 antagonist. Preincubation with an antibody against the IL-33 receptor ST2 inhibits (P< 0.0001) secretion of IL-1β stimulated either by IL-33 or together with SP. The combination of SP (1 μM) with IL-33 (30 ng/mL) increases IL-1β gene expression by 90-fold in LAD2 cells and by 200-fold in primary cultured mast cells from human umbilical cord blood. The combination of SP and IL-33 increases intracellular levels of IL-1β in LAD2 by 100-fold and gene expression of IL-1β and procaspase-1 by fivefold and pro-IL-1β by twofold. Active caspase-1 is present even in unstimulated cells and is detected extracellularly. Preincubation of LAD2 cells with the natural flavonoid methoxyluteolin (1–100 mM) inhibits (P< 0.0001) secretion and gene expression of IL-1β, procaspase-1, and pro-IL-1β. Mast cell secretion of IL-1β in response to SP and IL-33 reveals targets for the development of antiinflammatory therapies.

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Retinoic acid is a negative regulator for the differentiation of cord blood-derived human mast cell progenitors

Blood ◽

10.1182/blood.v95.9.2821.009k11_2821_2828 ◽

2000 ◽

Vol 95 (9) ◽

pp. 2821-2828 ◽

Cited By ~ 25

Author(s):

Tatsuya Kinoshita ◽

Kenichi Koike ◽

Hadija Hemed Mwamtemi ◽

Susumu Ito ◽

Shuichi Ishida ◽

...

Keyword(s):

Mast Cell ◽

Retinoic Acid ◽

Mast Cells ◽

Cord Blood ◽

Cell Development ◽

Negative Regulator ◽

Human Mast Cell ◽

Target Cells ◽

Dependent Manner ◽

Selective Agonist

We examined the effects of retinoids on the human mast cell development using a serum-deprived culture system. When 10-week cultured mast cells derived from CD34+ cord blood cells were used as target cells, both all-trans retinoic acid (ATRA) and 9-cis RA inhibited the progeny generation under stimulation with stem cell factor (SCF) in a dose-dependent manner (the number of progeny grown by SCF plus RA at 10−7 mol/L was one tenth of the value obtained by SCF alone). The early steps in mast cell development appear to be less sensitive to RA according to the single CD34+c-kit+ cord blood cell culture study. The optimal concentration of RAs also reduced the histamine concentration in the cultured mast cells (3.00 ± 0.47 pg per cell in SCF alone, 1.44 ± 0.18 pg per cell in SCF+ATRA, and 1.41 ± 0.10 pg per cell in SCF+9-cis RA). RT-PCR analyses showed the expression of RAR, RARβ, RXR, and RXRβ messenger ribonucleic acid (mRNA) in 10-week cultured mast cells. The addition of an RAR-selective agonist at 10−10 mol/L to 10−7 mol/L decreased the number of mast cells grown in SCF, whereas an RXR-selective agonist at up to 10−8 mol/L was inactive. Among RAR subtype selective retinoids used at 10−9 mol/L to 10−7 mol/L, only the RAR agonist was equivalent to ATRA at 10−7 mol/L in its ability to inhibit mast cell growth. Conversely, the addition of excess concentrations of a RAR antagonist profoundly counteracted the retinoid-mediated suppressive effects. These results suggest that RA inhibits SCF-dependent differentiation of human mast cell progenitors through a specific receptor.

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Expression of Functional TrkA Receptor Tyrosine Kinase in the HMC-1 Human Mast Cell Line and in Human Mast Cells

Blood ◽

10.1182/blood.v90.5.1807 ◽

1997 ◽

Vol 90 (5) ◽

pp. 1807-1820 ◽

Cited By ~ 105

Author(s):

See-Ying Tam ◽

Mindy Tsai ◽

Masao Yamaguchi ◽

Koji Yano ◽

Joseph H. Butterfield ◽

...

Keyword(s):

Mast Cell ◽

Mast Cells ◽

Tyrosine Kinase ◽

Cord Blood ◽

Umbilical Cord ◽

Umbilical Cord Blood ◽

Human Mast Cell ◽

Human Umbilical Cord Blood ◽

Human Umbilical Cord ◽

Human Mast Cells

Abstract Nerve growth factor (NGF ) can influence mast cell development and function in murine rodents by interacting with its receptors on mast cells. We now report the identification of mRNA transcripts of full-length tyrosine kinase-containing trkA, trkB, and trkC neurotrophin receptor genes in HMC-1 human mast cell leukemia cells. Although HMC-1 cells lacked p75 mRNA, they expressed transcripts for the exon-lacking splice variant of trkA (trkAI), truncated trkB (trkB.T1), and truncated trkC. By flow cytometry, HMC-1 cells exhibited expression of TrkA, TrkB, and TrkC receptor proteins containing full-length tyrosine kinase domains. NGF stimulation of HMC-1 cells induced tyrosine phosphorylation of TrkA protein, increased expression of the early response genes c-fos and NGF1-A, and activation of ERK-mitogen–activated protein (MAP) kinase, results which indicate that TrkA receptors in HMC-1 cells are fully functional. Highly purified populations of human lung mast cells expressed mRNAs for trkA, trkB and trkC, whereas preparations of human umbilical cord blood-derived mast cells expressed mRNAs for trkA and trkC, but not trkB. Moreover, preparations of human umbilical cord blood-derived immature mast cells not only expressed mRNA transcript and protein for TrkA, but exhibited significantly higher numbers of chymase-positive cells after the addition of NGF to their culture medium for 3 weeks. In addition, HMC-1 cells expressed mRNAs for NGF, brain-derived neurotrophic factor (BDNF ), and neurotrophin-3 (NT-3), the cognate ligands for TrkA, TrkB, and TrkC, whereas NGF and BDNF transcripts were detectable in human umbilical cord blood mast cell preparations. Taken together, our findings show that human mast cells express a functional TrkA receptor tyrosine kinase and indicate that NGF may be able to promote certain aspects of mast cell development and/or maturation in humans. Our studies also raise the possibility that human mast cells may represent a potential source for neurotrophins.

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Effects of early life adversity on meningeal mast cells and proinflammatory gene expression in male and female Mus musculus

10.1101/2021.09.17.460793 ◽

2021 ◽

Author(s):

Natalia Duque-Wilckens ◽

Erika Sarno ◽

Robby E. Teis ◽

Frauke Stoelting ◽

Sonia Khalid ◽

...

Keyword(s):

Gene Expression ◽

Mast Cell ◽

Immune System ◽

Mast Cells ◽

Early Life ◽

Inflammatory Markers ◽

Disease Susceptibility ◽

Early Life Adversity ◽

Male And Female ◽

The Brain

ABSTRACTExposure to early life adversity (ELA) in the form of physical and/or psychological abuse or neglect increases the risk of developing psychiatric and inflammatory disorders later in life. It has been hypothesized that exposure to ELA results in persistent, low grade inflammation that leads to increased disease susceptibility by amplifying the crosstalk between stress-processing brain networks and the immune system, but the mechanisms remain largely unexplored. The meninges, a layer of three overlapping membranes that surround the central nervous system (CNS)- duramater, arachnoid, and piamater – possess unique features that allow them to play a key role in coordinating immune trafficking between the brain and the peripheral immune system. These include a network of lymphatic vessels that carry cerebrospinal fluid from the brain to the deep cervical lymph nodes, fenestrated blood vessels that allow the passage of molecules from blood to the CNS, and a rich population of resident mast cells, master regulators of the immune system. Using a mouse model of ELA consisting of neonatal maternal separation plus early weaning (NMSEW), we sought to explore the effects of ELA on duramater mast cell histology and expression of inflammatory markers in male and female C57Bl/6 mice. We found that mast cell number, activation level, and relative expression of pseudopodia differ across duramater regions, and that NMSEW exerts region-specific effects on mast cells in males and females. Using gene expression analyses, we next found that NMSEW increases the expression of inflammatory markers in the duramater of females but not males, and that this is prevented by pharmacological inhibition of mast cells with ketotifen. Together, our results show that ELA drives sex-specific, long-lasting effects on the duramater mast cell population and immune-related gene expression, suggesting that the long-lasting effects of ELA on disease susceptibility could be partly mediated by meningeal function.

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Serial Analysis of Gene Expression in Human Cord Blood-Derived Mast Cells

The Japanese Journal of Pharmacology ◽

10.1016/s0021-5198(19)48080-3 ◽

2000 ◽

Vol 82 ◽

pp. 154

Author(s):

Atsuo Kuramasu ◽

Yasuhiko Kubota ◽

Hirohisa Saito ◽

Takehiko Watanabe ◽

Hiroshi Ohtsu

Keyword(s):

Gene Expression ◽

Mast Cells ◽

Cord Blood ◽

Human Cord Blood

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Data analysis-driven precise asthmatic treatment by targeting mast cells

Endocrine Metabolic & Immune Disorders - Drug Targets ◽

10.2174/1871530320666200610152922 ◽

2020 ◽

Vol 20 ◽

Author(s):

Yupin Tan ◽

Lili Zou ◽

Na Li ◽

Li Huang ◽

Meiji Chen ◽

...

Keyword(s):

Gene Expression ◽

Mast Cell ◽

Mast Cells ◽

Pulmonary Inflammation ◽

Functional Enrichment ◽

Global Changes ◽

Hub Gene ◽

Cell Engraftment ◽

Inhaled Budesonide ◽

Array Analyses

Background: Although importance of mast cells in asthma has been studied, mast cells-induced global changes in lungs are largely unknown. Data-driven identification contributes to discovering significant biomarkers or therapeutic targets, which are the basis of effective clinical medications. Objective: This study aims to explore the effects of mast cells on gene expression in asthmatic lungs, and to assess curative effects of inhaled budesonide (BUD). Methods and Results: Pulmonary gene expression in KitWsh mice with or without mast cell engraftment was analyzed with R software. Functional enrichment of Gene Ontology and KEGG were carried out through DAVID. Hub genes were identified with String and Cytoscape. The array analyses showed that the mast cell engraftment enhanced inflammation/immune response, cytokine/chemokine signal, and monocyte/neutrophil/lymphocyte chemotaxis. Interleukin (IL)-6 was identified to be a significant hub gene with the highest degree. Based on this, the effects of BUD were investigated on the aspects of anti-inflammation. BUD’s treatment was found to reduce serum IL-6 content and pulmonary inflammation in the ovalbumin-induced asthma rats. The treatment also downregulated beta-tryptase expression in both lung tissues and serum. Morphologically, the accumulation and degranulation of mast cells were significantly suppressed. Notably, the effects of BUD on inflammation and degranulation were comparable with tranilast (a classic mast cell inhibitor), while a synergy effect was not found. Conclusion: This study presented a unique pulmonary gene profile induced by mast cell engraftment, which could be reversed through blockage of mast cells or inhaled BUD.

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

Journal of Allergy and Clinical Immunology ◽

10.1016/s0091-6749(98)70064-8 ◽

1998 ◽

Vol 102 (1) ◽

pp. 134-142 ◽

Cited By ~ 23

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

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GATA-Dependent Expression of the Interleukin-1 Receptor-Related T1 Gene in Mast Cells

Molecular and Cellular Biology ◽

10.1128/mcb.18.9.5320 ◽

1998 ◽

Vol 18 (9) ◽

pp. 5320-5331 ◽

Cited By ~ 21

Author(s):

Thomas Gächter ◽

Dirk R. Moritz ◽

Jaqueline Gheyselinck ◽

Roman Klemenz

Keyword(s):

Gene Expression ◽

Mast Cell ◽

Mast Cells ◽

Mutational Analysis ◽

Interleukin 1 ◽

Point Mutations ◽

Calcium Ionophore ◽

Normal Growth ◽

Promoter Strength ◽

T1 Gene

ABSTRACT The murine delayed-early serum-responsive gene T1 encodes glycoproteins of the interleukin-1 receptor family. Transcriptional initiation in fibroblasts is regulated by c-Fos and gives rise to a rare 5-kb mRNA and an abundant 2.7-kb mRNA. These transcripts are translated into a receptor-like membrane-anchored protein and a secreted protein consisting only of the ectodomain. In mast cells, T1 gene transcription is initiated 10.5 kb further upstream than in fibroblasts and gives rise predominantly to the 5-kb transcript under normal growth conditions. Here we demonstrate that calcium ionophore stimulation of mast cells resulted in an upregulation of T1 gene expression and a switch from the long to the short T1 transcript. This was paralleled by the disappearance of the receptor-type T1 protein on the mast cell surface and the secretion of large amounts of the truncated T1 protein. c-Fos and a T1 enhancer, which have previously been identified to be essential for T1 expression in fibroblasts, were not required for calcium ionophore-mediated T1 gene upregulation. Overexpression of the transcription factor GATA-1 in mast cells caused elevated T1 synthesis. Three GATA elements were identified in the minimal GATA-responsive mast cell promoter. Mutational analysis revealed that all three GATA elements are involved in T1 gene expression. Point mutations within the middle GATA element eliminated promoter activity completely, while mutations of the distal and proximal GATA binding sites reduced promoter strength by factors of 2 and 5, respectively. Exogenous expression of GATA-1 was not sufficient to activate the mast cell-specific promoter in NIH 3T3 fibroblasts.

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Interleukin-6 Directly Modulates Stem Cell Factor-Dependent Development of Human Mast Cells Derived From CD34+Cord Blood Cells

Blood ◽

10.1182/blood.v94.2.496.414k19_496_508 ◽

1999 ◽

Vol 94 (2) ◽

pp. 496-508 ◽

Cited By ~ 1

Author(s):

Tatsuya Kinoshita ◽

Nobukuni Sawai ◽

Eiko Hidaka ◽

Tetsuji Yamashita ◽

Kenichi Koike

Keyword(s):

Mast Cell ◽

Stem Cell ◽

Mast Cells ◽

Cord Blood ◽

Interleukin 6 ◽

Blood Cells ◽

Stem Cell Factor ◽

Flow Cytometric Analysis ◽

Flow Cytometric ◽

Cord Blood Cells

In the present study, we attempted to clarify the effects of interleukin-6 (IL-6) on the growth and properties of human mast cells using cultured mast cells selectively generated by stem cell factor (SCF) from CD34+ cord blood cells. The addition of IL-6 to cultures containing mast cells resulted in a substantial reduction of the number of progenies grown by SCF in the liquid culture. This IL-6–mediated inhibition of mast cell growth may be due in part to the suppression at the precursor level, according to the results of a clonal cell culture assay. Moreover, a flow cytometric analysis showed that the cultured mast cells grown in the presence of SCF+IL-6 had decreased c-kit expression. The exposure of cultured mast cells to SCF+IL-6 also caused substantial increases in the cell size, frequency of chymase-positive cells, and intracellular histamine level compared with the values obtained with SCF alone. The flow cytometric analysis showed low but significant levels of expression of IL-6 receptor (IL-6R) and gp130 on the cultured mast cells grown with SCF. The addition of either anti–IL-6R antibody or anti-gp130 antibody abrogated the biological functions of IL-6. Although IL-4 exerted an effect similar to that of IL-6 on the cultured mast cells under stimulation with SCF, the results of comparative experiments suggest that the two cytokines use different regulatory mechanisms. Taken together, the present findings suggest that IL-6 modulates SCF-dependent human mast cell development directly via an IL-6R-gp130 system.

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