Basic and Clinical ImmunologyIdentification of granulocyte subtype–selective receptors and ion channels by using a high-density oligonucleotide probe array☆
Section snippets
Purification of leukocytes
All human subjects in this study provided written informed consent, and the ethical review boards at the relevant hospitals (National Center for Child Health and Development and Jikei University School of Medicine) approved the study. The subjects used in this study were all healthy volunteers, specifically chosen for having no allergic diseases.
Granulocytes and mononuclear cells were separated from the venous blood of normal volunteers. Human basophils were semipurified by means of Percoll
Results
In this study we have used a high-density oligonucleotide probe array (GeneChip) to measure the expression levels of approximately 20,000 different transcripts in highly purified cells. These cells were basophils, eosinophils, neutrophils, monocytes (CD14+), T lymphocytes (CD4+ and CD8+ cells), B lymphocytes (CD19+), lung-derived mast cells, cord blood–derived cultured mast cells, and nasal polyp–derived fibroblasts. The GeneChip assay allows the simultaneous measurement of large numbers of
Discussion
We identified 51 granulocyte-selective genes for ion channels and receptors by examining approximately 20,000 kinds of transcripts derived from 16,000 genes from 10 different types of cells with the U133A GeneChip, which covers approximately half of the genes present in the human genome. The majority of these transcripts encoded molecules known or expected to be granulocyte subtype–selective, such as the IL-3 receptor and Fcε receptors.
Mast cells expressed low levels of FcεRIα compared with
Acknowledgements
We thank Ms Noriko Hashimoto, Mr Keisuke Yuki, Mr Hisashi Tomita, Mr Nobuyuki Baba, and Ms Nao Aida at The National Research Institute for Child Health and Development for discussion and skillful technical assistance. We also thank Mr Igor A. Bagayev at The Confocal Microscope Facility, Massachusetts Genral Hospital, Harvard Medical School.
References (39)
Systemic activation of basophils and eosinophils: markers and consequences
J Allergy Clin Immunol
(2000)- et al.
Polytrauma induces increased expression of pyruvate kinase in neutrophils
Blood
(2000) - et al.
G protein-coupled receptors
I. Diversity of receptor-ligand interactions. J Biol Chem
(1998) - et al.
Selective down-regulation of high-affinity IgE receptor (FcηRI) α-chain messenger RNA among transcriptome in cord blood-derived versus adult peripheral blood-derived cultured human mast cells
Blood
(2001) - et al.
Gene expression screening of human mast cells and eosinophils using high-density oligonucleotide probe arrays: abundant expression of major basic protein in mast cells
Blood
(2001) - et al.
Marked increase in CC chemokine gene expression in both human and mouse mast cell transcriptomes following Fc∗ receptor I cross-linking: an interspecies comparison
Blood
(2002) - et al.
Glucocorticoids induce basophil apoptosis
J Allergy Clin Immunol
(2001) - et al.
Induction of apoptosis in human eosinophils by anti-Fas antibody treatment in vitro
Blood
(1995) - et al.
Assessment of the anti-c-kit monoclonal antibody YB5.B8 in affinity magnetic enrichment of human lung mast cells
J Immunol Methods
(1994) - et al.
Ligation of Siglec-8: a selective mechanism for induction of human eosinophil apoptosis
Blood
(2003)
IL-4 production by human basophils found in the lung following segmental allergen challenge
J Allergy Clin Immunol
IgE(+), Kit(-), I-A/I-E(-) myeloid cells are the initial source of Il-4 after antigen challenge in a mouse model of allergic pulmonary inflammation
J Allergy Clin Immunol
Regulation of the function of eosinophils and basophils
Crit Rev Immunol
The evolving pathogenesis of systemic vasculitis
Clin Med
Characterization of histamine secretion from mechanically dispersed human lung mast cells: effects of anti-IgE, calcium ionophore A23187, compound 48/80, and basic polypeptides
J Immunol
Voltage-gated potassium channels regulate calcium-dependent pathways involved in human T lymphocyte activation
J Exp Med
Effects of ADP-ribosylation of GTP-binding protein by pertussis toxin on immunoglobulin E-dependent and -independent histamine release from mast cells and basophils
J Immunol
Knockouts model the 100 best-selling drugs
Will they model the next 100? Nat Rev Drug Discov
The sequence of the human genome
Science
Cited by (46)
IL-33 induces functional CCR7 expression in human mast cells
2018, Journal of Allergy and Clinical ImmunologyHuman eosinophils constitutively express a unique serine protease, PRSS33
2017, Allergology InternationalCitation Excerpt :Recombinant human serine protease 33 (PRSS33) was synthesized at Sysmex Corporation (Kanagawa, Japan) using a baculovirus gene expression system based on the reference sequence (NM_152891.2). Each type of leukocyte was isolated from peripheral blood of both healthy and mildly allergic donors (n = 10) by density gradient sedimentation using Lymphocyte Separation Medium (Wako Pure Chemical Industries, Osaka, Japan) or Percoll (GE Healthcare, Piscataway, NJ), and also by positive and/or negative selection using immunomagnetic beads (Miltenyi Biotec, Bergisch-Gladbach, Germany), as described previously.11,16 The purity of eosinophils based on light microscopic examination of cytocentrifuge preparations using Cytospin (Shandon, Pittsburgh, PA) and staining with Diff-Quik (American Scientific Products, McGraw Park, IL) always exceeded 98%.
Microbiome and Allergy
2016, Encyclopedia of ImmunobiologyMast Cells and IgE: From history to today
2013, Allergology InternationalContribution of histamine metabolism to tachyphylaxis during the buildup phase of rush immunotherapy
2009, Journal of Allergy and Clinical Immunology
- ☆
Supported in part by a grant from the Organization for Pharmaceutical Safety and Research and the Ministry of Health, Labour and Welfare (the Millennium Genome Project, MPJ-5), a grant from RIKEN Research Center for Allergy and Immunology (to H.S.), National Institutes of Health grant AI 43663 from the National Institute of Allergy and Infectious Diseases, grant RSG-01-241-01-LIB from the American Cancer Society (to C.A.), and by the Adra family.