Surface expression of HLA-DM on dendritic cells derived from CD34-positive bone marrow haematopoietic stem cells.
HLA-DM has been known to be largely absent from the cell surface of antigen-presenting cells, accumulating instead in the intracellular compartment. In this study, we demonstrated that a population of HLA-DM-positive (HLA-DM+) dendritic cells (DCs) can be identified in an in vitro culture of CD34+ bone marrow haematopoietic stem cells. CD34+ bone marrow cells of healthy donors were used to generate DCs with the recombinant human cytokines granulocyte-macrophage colony-stimulating factor (GM-CSF), tumour necrosis factor alpha (TNF-alpha) and stem cell factor (SCF), both with and without interleukin 4 (IL-4). Flow cytometric analysis demonstrated that HLA-DM+ cells comprised 2.5 +/- 0.9% and 1.8 +/- 0.4% of the CD34+ cell-derived progeny in the presence of GM-CSF, TNF-alpha and SCF after 7 d and 14 d of culture respectively. The number of HLA-DM molecules expressed per HLA-DM+ cell on d 7 was significantly higher than that on d 14 (1410 +/- 47 versus 370 +/- 25, P < 0.05). The addition of IL-4 to the cytokines from the commencement of culture increased the proportion of HLA-DM+ cells and increased the number of HLA-DM molecules per HLA-DM+ cell significantly (P < 0.05). Although most of the HLA-DM+ cells expressed CD1a, CD80 or CD86 antigen, only a small proportion of CD1a+, CD80+ or CD86+ cells expressed HLA-DM. About half the HLA-DM+ cells expressed CD83. The addition of IL-4 resulted in a decrease in the expression of CD83 on the HLA-DM+ cells on d 7. Microscopic evaluations of sorted HLA-DM+ cells revealed the characteristic morphological features of DCs. Primary mixed lymphocyte cultures demonstrated that the HLA-DM+ cells elicited a vigorous proliferation of allogeneic T cells. The level of antigen-specific T-cell activation induced by antigen-pulsed, chloroquine-treated HLA-DM+ cells was substantially higher than that induced by HLA-DM- cells (P < 0.05). These results show that HLA-DM can be used as a useful DC lineage-specific marker, as well as a tool for the characterization of DCs and human immunotherapy.
Min
Y H
YH
Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea. minbrmmd@yumc.yonsei.ac.kr
Lee
S T
ST
Choi
K M
KM
Hahn
J S
JS
Ko
Y W
YW
D018949:Antigens, CD1 / Q000276:immunology; D018952:Antigens, CD34 / Q000276:immunology*; D018122:B7-1 Antigen / Q000276:immunology; D002455:Cell Division; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology*; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000276:immunology; D006681:HLA-D Antigens / Q000276:immunology*; D006412:Hematopoietic Stem Cells / Q000276:immunology*; D006801:Humans; D015847:Interleukin-4 / Q000276:immunology; D008214:Lymphocytes / Q000166:cytology; D019089:Stem Cell Factor / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
D018949:Antigens, CD1; D018952:Antigens, CD34; D018122:B7-1 Antigen; D006681:HLA-D Antigens; D019089:Stem Cell Factor; D014409:Tumor Necrosis Factor-alpha; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor