Selective down-regulation of high-affinity IgE receptor (FcεRI) α-chain messenger RNA among transcriptome in cord blood–derived versus adult peripheral blood–derived cultured human mast cells

Abstract

Substantial numbers of human mast cells (MCs) were generated from umbilical cord blood (CB) and from adult peripheral blood (PB). A single CB progenitor produced 15 436 MCs, whereas a single PB progenitor produced 807 MCs on average. However, PB-derived MCs were far more active than CB-derived MCs in terms of high-affinity IgE receptor (FcepsilonRI)-mediated reactions. One million sensitized PB-derived MCs released 3.6 microg histamine, 215 pg IL-5, and 14 ng granulocyte macrophage-colony-stimulating factor (GM-CSF), whereas 10(6) sensitized CB-derived MCs released only 0.8 microg histamine, 31 pg IL-5, and 0.58 ng GM-CSF on anti-IgE challenge. However, ionophore A23 187 released similar levels of histamine from the 2 MC types. PB-derived MCs highly expressed surface FcepsilonRI alpha chain, and CB-derived MCs almost lacked it in the absence of IgE. PB-derived MCs expressed approximately 5 times higher levels of messenger RNA (mRNA) for FcepsilonRI alpha chain than CB-derived MCs, but mRNAs for beta and gamma chains of the receptors were equally expressed. Among the approximately 5600 kinds of full-length human genes examined by using the high-density oligonucleotide probe-array system, FcepsilonRIalpha was ranked the fifth most increased transcript in PB-derived MCs. The 4 other increased transcripts were unrelated to MC function. These results suggest that IgE-mediated reactions may be restricted during early infancy through the selective inhibition of FcepsilonRIalpha transcription, which is probably committed at progenitor stages and is, at least in part, cytokine-insensitive.

Redefinition of the human mast cell transcriptome by deep-CAGE sequencing

“…Apart from several exceptions, there was good overall agreement with previous profiling efforts, implying that this pathway is fairly consistent among MC subsets. [12][13][14][15][16] IL-31 was appreciably expressed in only one of the stimulated MC samples, suggesting particularly tight control. Testing MCs from 10 individuals, we found highly variable IL-31 protein levels among donors (supplemental Figure 2B).…”

“…11 Research into the full range of functional programs of a cell requires information on its specific gene-activity profile. Previous transcriptome profiling efforts concentrated either on MCs alone (eg, stimulated vs baseline [12][13][14] and MCs cultured from different sources 15,16 or at different times of development 17 ) or on comparisons between MCs and other leukocytes (either experimentally 18 or in silico 19 ). Although these studies provided valuable insights into the transcriptional landscape of MCs, they did not allow direct comparisons with nonimmune cells and were limited by the MC model(s) employed.…”

Gene expression screening of human mast cells and eosinophils using high-density oligonucleotide probe arrays: abundant expression of major basic protein in mast cells

“…The cells were mixed well with 2.7 mL serum-free Iscoves methylcellulose medium (Stem Cell Technologies, Vancouver, BC, Canada) supplemented with 200 ng/mL SCF, 50 ng/mL IL-6, and 1 ng/mL IL-3, as described previously. 20 The cell suspension was inoculated at 0.3 mL per well to 24-well plates (Iwaki Glass, Tokyo, Japan) at 37°C in 5% CO 2 . Every 2 weeks, 0.3 mL fresh methylcellulose medium containing 100 ng/mL SCF and 50 ng/mL IL-6 was layered over the methylcellulose cultures.…”

“…The competition with another cell type required for cDNA microarray assay is not required with the Genechip. 16,[18][19][20] Thus, we can compare the expression levels of more than 10 000 transcripts even in different cell types by using the high-density oligonucleotide probe array. In the present study, we used the Genechip to measure and compare cultured human MCs and purified eosinophils for their expression levels of more than 10 000 transcripts.…”

T Cell Proliferation by Direct Cross-Talk between OX40 Ligand on Human Mast Cells and OX40 on Human T Cells: Comparison of Gene Expression Profiles between Human Tonsillar and Lung-Cultured Mast Cells

“…For stimulation of MCs via C5aR, MCs were challenged with 3 ϫ 10 Ϫ8 M C5a (Sigma-Aldrich, St. Louis, MO) for 30 min for histamine assay. For histamine release experiments, the cells were suspended in Tyrode solution as described previously (31). In other experiments, the cells were suspended in complete IMDM containing rSCF and rIL-6.…”