title: Reduced expression of granule proteins during extended survival of eosinophils in splenocyte culture with GM-CSF.
abstract: Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a multifaceted hematopoietic cytokine and the culture of mouse bone marrow with GM-CSF produces a variety of myeloid cells including granulocytes, macrophages, and dendritic cells. In the present study, we cultured mouse splenocytes with GM-CSF and examined the changes in hematopoietic cell populations over a week. Most of the splenic hematopoietic cells disappeared significantly from culture within 6days with or without the presence of GM-CSF. Among the splenic granulocyte populations, only eosinophils fully survived throughout the culture with GM-CSF for more than a week. During 10days of culture with GM-CSF, splenic eosinophils maintained their morphology as well as most of their surface molecules at high levels, including CCR3 and Siglec F. Meanwhile, the expression of mRNAs encoding major basic protein-1 (MBP-1) and eosinophil peroxidase (EPO), two major eosinophil-derived granule proteins, was diminished significantly from the cultured eosinophils. EPO assays also revealed that eosinophils in culture for more than 5days retained 30% or less EPO activity compared to those in uncultured splenocytes. In contrast, culture of splenocytes with GM-CSF did not change the capacity of eosinophils to migrate in response to eotaxin-1. Our results indicate that mouse splenic eosinophils are effectively cultured for lengthy periods while their expression of eosinophil-derived granule proteins is specifically suppressed. The relevance of these findings to eosinophilic inflammatory response is discussed.
authors.affiliation: Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Seul Hye
authors.initials: SH
authors.lastname: Ryu
authors.affiliation: Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Hye Young
authors.initials: HY
authors.lastname: Na
authors.affiliation: Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Moah
authors.initials: M
authors.lastname: Sohn
authors.affiliation: Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Sun Murray
authors.initials: SM
authors.lastname: Han
authors.affiliation: Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Wanho
authors.initials: W
authors.lastname: Choi
authors.affiliation: Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Hyunju
authors.initials: H
authors.lastname: In
authors.affiliation: Laboratory of Molecular Virology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Sookyung
authors.initials: S
authors.lastname: Hong
authors.affiliation: Laboratory of Molecular Virology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Hyejin
authors.initials: H
authors.lastname: Jeon
authors.affiliation: Laboratory of Molecular Virology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Jun-Young
authors.initials: JY
authors.lastname: Seo
authors.affiliation: WOOJUNG Life Science Research Center, WOOJUNGBSC, Suwon, Gyeonggi-do 16229, Republic of Korea.
authors.forename: Jongcheol
authors.initials: J
authors.lastname: Ahn
authors.affiliation: Laboratory of Immunology, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul 03722, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Republic of Korea. Electronic address: ChaeGyu@yuhs.ac.
authors.forename: Chae Gyu
authors.initials: CG
authors.lastname: Park
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002465:Cell Movement; D002470:Cell Survival; D002478:Cells, Cultured; D054413:Chemokine CCL11 / Q000276:immunology; D003594:Cytoplasmic Granules / Q000378:metabolism; D047093:Eosinophil Major Basic Protein / Q000235:genetics / Q000378:metabolism; D047209:Eosinophil Peroxidase / Q000235:genetics / Q000378:metabolism; D004804:Eosinophils / Q000276:immunology*; D005786:Gene Expression Regulation; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism*; D006410:Hematopoiesis; D051379:Mice; D008810:Mice, Inbred C57BL; D054397:Receptors, CCR3 / Q000378:metabolism; D063267:Sialic Acid Binding Ig-like Lectin 1 / Q000378:metabolism; D013154:Spleen / Q000276:immunology* / Q000473:pathology
chemical_list: C515351:Ccr3 protein, mouse; D054413:Chemokine CCL11; D054397:Receptors, CCR3; D063267:Sialic Acid Binding Ig-like Lectin 1; C571167:Siglec1 protein, mouse; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D047209:Eosinophil Peroxidase; D047093:Eosinophil Major Basic Protein
keywords: Eosinophils; GM-CSF; Gene expression; Granule protein; Mouse; Spleen