Basic and Clinical Immunology
Identification of granulocyte subtype–selective receptors and ion channels by using a high-density oligonucleotide probe array

https://doi.org/10.1016/j.jaci.2003.12.036Get rights and content

Abstract

Background

During inflammation, neutrophils, basophils, and eosinophils release cell type–specific mediators and proteases through signaling molecules, such as G protein–coupled receptors and ion channels. As such, ion channels and receptors, including G protein–coupled receptors, are common drug targets.

Objective

We sought to identify, for the first time, ion channels and receptors preferentially expressed by each granulocyte subtype.

Methods

Using GeneChip, we compared approximately 20,000 transcripts present in 7 leukocyte types, platelets, mast cells, and fibroblasts to identify granulocyte subtype–selective transcripts for receptors and ion channels. Granulocyte subtype–selective transcripts were chosen on the basis of several conditions, such as the transcript having a 5-fold or greater expression level compared with the maximum level of other leukocytes.

Results

Fifty-one transcripts were chosen to be preferentially expressed by each granulocyte subtype. Seventeen of the 51 transcripts have not been previously reported as granulocyte subtype selective. Among the 17 receptors and ion channels, 6 were basophil selective, eosinophil selective, or both and were not highly expressed by other organs, indicating that they might be potential targets for antiallergy drugs.

Conclusion

Use of this database of potential cell type–selective drug targets should minimize the efforts required for pharmaceutical development.

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.

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    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.

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