Differentiation of a human eosinophilic leukemic cell line, EoL-1: characterization by the expression of cytokine receptors, adhesion molecules, CD95 and eosinophilic cationic protein (ECP).
Purification of enough eosinophils for the study of allergic inflammation is difficult because eosinophils comprise only a small percentage of circulating leucocytes. A human eosinophilic leukemic cell line, EoL-1, has been considered to be an in vitro eosinophilic model. In the present study, the suitability of EoL-1 cells as an eosinophilic model was further investigated. EoL-1 cells were induced to differentiate by dibutyryl cyclic AMP (dbcAMP). The expression of cell surface cytokines (IL-3, IL-5, GM-CSF) receptors, adhesion molecules (CD49d, CD11b), and CD95 (Fas) was investigated by flow cytometry. Expression of eosinophilic cationic protein (ECP) was determined by fluorescence enzyme immunoassay (FEIA) and reverse transcription-polymerase chain reaction (RT-PCR). EoL-1 cells could be differentiated into eosinophilic vacuole-containing cells by dbcAMP. They were found to express cell surface IL-3 and GM-CSF receptors, CD95 and CD49d. Treatment with dbcAMP could induce the expression of CD11b but decrease the expression of CD95. Anti-CD95 antibody could induce their apoptosis. The differentiation of EoL-1 cells was accompanied by increase in release of ECP into the supernatant and total ECP synthesis. Differentiation of EoL-1 cells also up-regulated the expression of mRNA for ECP and its level was parallel to the total amount of ECP synthesis. By virtue of their expression of haematopoietic cytokines receptors, adhesion molecules, CD95, synthesis and release of ECP, EoL-1 cells are suitable as an in vitro eosinophilic model for studying eosinophilic functions.
Wong
C K
CK
Department of Chemical Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin NT, PR China.
Ho
C Y
CY
Lam
C W
CW
Zhang
J P
JP
Hjelm
N M
NM
D015703:Antigens, CD / Q000032:analysis; D000943:Antigens, Differentiation; D017209:Apoptosis; D001798:Blood Proteins / Q000032:analysis / Q000235:genetics; D003994:Bucladesine / Q000494:pharmacology; D015815:Cell Adhesion Molecules / Q000032:analysis; D002454:Cell Differentiation; D047091:Eosinophil Granule Proteins; D004804:Eosinophils / Q000166:cytology*; D006801:Humans; D017681:Hypereosinophilic Syndrome; D039441:Integrin alpha4; D016177:Macrophage-1 Antigen / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D018121:Receptors, Cytokine / Q000032:analysis; D012260:Ribonucleases; D014407:Tumor Cells, Cultured; D019014:fas Receptor / Q000032:analysis
D015703:Antigens, CD; D000943:Antigens, Differentiation; D001798:Blood Proteins; D015815:Cell Adhesion Molecules; D047091:Eosinophil Granule Proteins; D016177:Macrophage-1 Antigen; D012333:RNA, Messenger; D018121:Receptors, Cytokine; D019014:fas Receptor; D039441:Integrin alpha4; D003994:Bucladesine; D012260:Ribonucleases