[The biological characteristics of peripheral blood CD123+ myeloid dendritic cell].
OBJECTIVE
To investigate the biological characteristics of CD123(+)DC derived from cord blood monocytes in myeloid culture system.


METHODS
The monocytes isolated from peripheral blood were cultured with GM-CSF and IL-4 to induce DC for 7 days, and the related phenotype of DC was analyzed by flow cytometery. CD123(+)DC were purified with magnetic cell sorting separation technique. The morphological properties of CD123(+)DC was observed under confocal laser-scanning microscope and scan electron microscope. The concentration of IL-12 secreted by DC was analyzed by ELISA. The function of endocytosis and antigen presentation was evaluated by uptaking FITC-dextran and allogeneic mixed T lymphocyte reaction (MLR).


RESULTS
FACS analysis showed the high expression of CD86 and CD11C, moderate expression of CD1a and CD123, low levels of CD83 on DC after induction by GM-CSF and IL-4 for 7 days, while the expression of CD304 was rather low. Well-distributed CD123 and CD11C were detected on DC. Purified CD123(+)DC showed typical morphology of immature DC with similar short cytoplasmic projections as CD123(-)DC. CD123(+)DC secreted lower level of IL-12, and exhibited more significant endocytosis and less antigen presentation function than that of CD123(-)DC(P<0.05).


CONCLUSIONS
CD123(+)DC induced with GM-CSF and IL-4 might be more early phase of immature myeloid DC along with the DC differentiation, with special biological characteristics.
Shi
Jun
J
Department of Hematology, Sixth People's Hospital of Shanghai Jiaotong University, Shanghai, China. shijun7@hotmail.com
Wang
Min
M
Wan
Yun
Y
Ikeda
Kazuma
K
D017951:Antigen Presentation; D015703:Antigens, CD / Q000378:metabolism; D018949:Antigens, CD1 / Q000378:metabolism; D051940:B7-2 Antigen / Q000378:metabolism; D039521:CD11c Antigen / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D004705:Endocytosis; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism; D053650:Interleukin-3 Receptor alpha Subunit / Q000378:metabolism*; D015847:Interleukin-4 / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism*; D007959:Lymphocyte Culture Test, Mixed; D008562:Membrane Glycoproteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D022423:Myeloid Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000097802:CD83 Antigen
D015703:Antigens, CD; D018949:Antigens, CD1; D051940:B7-2 Antigen; D039521:CD11c Antigen; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D007136:Immunoglobulins; D018664:Interleukin-12; D053650:Interleukin-3 Receptor alpha Subunit; D015847:Interleukin-4; D008562:Membrane Glycoproteins; D000097802:CD83 Antigen; C419393:CD1a antigen; C497964:CD86 protein, human; C115022:IL3RA protein, human