A cell line model for the differentiation of human dendritic cells.

Abstract

We have identified human monocytic (THP-1) and myelogenous CD34+ (KG-1) leukemia cell lines that can be differentiated rapidly into mature dendritic cells (DCs) when cultured in serum-free medium containing GM-CSF, TNF-alpha, and ionomycin. These hematopoietic cell line-derived DCs are highly pure and monotypic, and display the morphologic, phenotypic, molecular, and functional properties of DCs generated from human donor-derived monocytes or CD34+ hematopoietic progenitor cells. During differentiation into mature DCs, the cells exhibit de novo cell-surface expression of CD83, CD80, CD86, CD40, CD206, CD209, CD120a, CD120b, and intracellular synthesis of IL-10, increase their endocytotic capacity, and acquire characteristic stellate morphology. To further define the cells as DCs, cytosolic induction and upregulation of RelB and RelA (p65), transcription factors of the NF-kappaB/Rel family essential for differentiation and maturation of DCs, as well as upregulation of the immunoproteasome subunits LMP2, LMP7, and MECL-1, and the proteasome activator PA28alpha, components essential for efficient MHC class I peptide antigen processing, were demonstrated during differentiation of the cells. In contrast to the cell lines, the cell line-derived mature DCs are capable of stimulating allogeneic CD4+ and CD8+ T cells, ultimately defining them as potent antigen-presenting cells. The approach to differentiate THP-1 and KG-1 cells into immature and mature DCs may serve as an experimental model to study molecular events and pathways that govern the differentiation of human malignant myeloid precursors, monocytes, and CD34+ hematopoietic progenitor cells into DCs.

• Authors: Carsten Berges, Cord Naujokat, Sarah Tinapp, Hubert Wieczorek, Alexandra Höh, Mahmoud Sadeghi, Gerhard Opelz, Volker Daniel
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 333 Issue 3 Pg. 896-907 (Aug 05 2005) ISSN: 0006-291X [Print] United States
• PMID: 15963458 (Publication Type: Journal Article)

Chemical References

• Antigens, CD34
• Culture Media, Serum-Free
• Lipopolysaccharide Receptors

Topics

• Antigens, CD34 (immunology)
• Cell Line
• Culture Media, Serum-Free
• Dendritic Cells (cytology, immunology)
• Flow Cytometry
• Humans
• Lipopolysaccharide Receptors (immunology)
• Models, Biological
• Polymerase Chain Reaction
• Reproducibility of Results