Human umbilical vein endothelial cells support interleukin-3- and interleukin-5-induced eosinophil differentiation from cord blood CD34+ cells.
BACKGROUND
Human umbilical vein endothelial cells (HUVEC) are an important source of hematopoietic cytokines, and interleukin-3 (IL-3)- and IL-5-induced eosinophil differentiation from CD34+ cells has been observed. To show the supportive effects of endothelial cells on eosinophil differentiation, we examined the effects of cocultured HUVEC on IL-3 and IL-5-induced eosinophil differentiation from human umbilical cord blood CD34+ cells.


METHODS
CD34+ cells were obtained from the heparinized umbilical vein blood of 10 volunteers using a CD34-conjugated magnetic bead positive direct selection procedure. With HUVEC in Transwell, CD34+ cells were then cultured for 14-28 days. In neutralizing experiments on HUVEC-derived cytokines, antibodies to both stem cell factor (SCF) and granulocyte/macrophage colony-stimulating factor (GM-CSF) were added to the cell cultures.


RESULTS
Cocultured HUVEC upregulated IL-3 and IL-5-induced eosinophil differentiation from CD34+ cells on day 28 of culture by 75.0%. The eosinophilopoietic effect of HUVEC was significantly only when the cells were present in the culture from day 15 to day 28. Addition of anti-SCF antibody or anti-GM-CSF monoclonal antibody to the culture significantly suppressed HUVEC-combined IL-3- and IL-5-induced eosinophil differentiation on day 28 of culture by 49.2 and 55.0%, respectively.


CONCLUSIONS
These results indicate that several cytokines including GM-CSF and SCF from HUVEC promote IL-3- and IL-5-induced eosinophil differentiation from CD34+ cells.
Ochiai
K
K
Department of Internal Medicine, Toho University School of Medicine, Sakura Hospital, Sakura, Japan. kysotiai@alles.or.jp
Omura
M
M
Mochizuki
A
A
Ito
M
M
Tomioka
H
H
D018952:Antigens, CD34; D002450:Cell Communication / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D004357:Drug Synergism; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology; D004804:Eosinophils / Q000166:cytology*; D005312:Fetal Blood; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology*; D015848:Interleukin-5 / Q000494:pharmacology*
D018952:Antigens, CD34; D007377:Interleukin-3; D015848:Interleukin-5