Role of interleukin-3 and signaling pathways on beta-thalassemia/HbE erythroid progenitor cell in culture.
In order to study the role of the cytokine interleukin-3 (IL-3) and its signaling pathways in erythropoiesis of beta-thalassemia/HbE erythroid progenitor cells, CD34 positive cells were isolated from peripheral blood of patients and healthy subjects. After culturing the cells in the presence or absence of IL-3, cell viability was measured by trypan blue staining and apoptotic cells were analyzed by flow cytometry. After 7 days of culture the highest percent erythroid progenitor cell viability was obtained with cells from healthy subjects, while the lowest percentage was found in those from splenectomized beta-thalassemia/HbE. Viability of beta-thalassemia/HbE erythroid progenitor cells in the presence of IL-3 was higher than that of nonsupplemented cells. In addition, specific inhibitors of protein kinase C (Ro-318220), phospholipase C (U-73122) and Janus kinase 2 (AG-490) were used to investigate the involvement of signaling pathways in erythropoiesis. Percent apoptosis of erythroid progenitor cells from splenectomized beta-thalassemia/HbE subjects treated with RO-318220 was higher than those of nonsplenectomized beta-thalassemia/HbE and healthy subjects. Treatment with U-73122 resulted in enhanced percent apoptotic cells from normal and beta-thalassemia/HbE subjects. All these effects were independent of IL-3 treatment.
Tanyong
Dalina I
DI
Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand. mtdic@mahidol.ac.th
Winichagoon
Pranee
P
Siripin
Darin
D
Seevakool
Weerayut
W
Fucharoen
Suthat
S
D000293:Adolescent; D000328:Adult; D018952:Antigens, CD34 / Q000097:blood / Q000276:immunology; D017209:Apoptosis / Q000276:immunology; D002648:Child; D015672:Erythroid Precursor Cells / Q000187:drug effects* / Q000276:immunology / Q000473:pathology; D004920:Erythropoiesis / Q000187:drug effects / Q000276:immunology*; D004963:Estrenes / Q000494:pharmacology; D005260:Female; D006446:Hemoglobin E / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007377:Interleukin-3 / Q000276:immunology* / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D011760:Pyrrolidinones / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D013154:Spleen / Q000276:immunology; D013156:Splenectomy; D017086:beta-Thalassemia / Q000097:blood* / Q000276:immunology
D018952:Antigens, CD34; D004963:Estrenes; D007377:Interleukin-3; D011760:Pyrrolidinones; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D006446:Hemoglobin E; D011493:Protein Kinase C