Expression of different NF-κB pathway genes in dendritic cells (DCs) or macrophages assessed by gene expression profiling

Ioannis Baltathakis, Orlando Alcantara, David H. Boldt

Research output: Contribution to journalArticlepeer-review

59 Scopus citations


NF-κB/Rel transcription factors have been implicated in the differentiation of monocytes to either dendritic cells (DCs) or macrophages, as well as in the maturation of DCs from antigen-processing to antigen-presenting cells. Recent studies of the expression pattern of Rel proteins and their inhibitors (IκBs) suggest that their regulation during this differentiation process is transcriptional. To investigate differential gene expression between macrophages and DCs, we used commercially available gene microarrays (GEArray KIT), which included four of the NF-κB/Rel family genes (p50/p105, p52/p100, RelB, and c-rel) and 32 additional genes either in the NF-κB signal transduction pathway or under transcriptional control of NF-κB/Rel factors. To generate macrophages and DCs, human adherent peripheral blood monocytes were cultured with M-CSF or GM-CSF + IL-4 respectively for up to 8 days. DCs (and in some experiments, macrophages) were treated with lipopolysaccharide (LPS) for the last 48 h of culture to induce maturation. Cells were harvested after 7 days, cDNA was prepared and radiolabeled with α-32P-dCTP, then hybridized to gene arrays containing specific gene probes. β-actin and GAPDH or PUC18 oligonucleotides served as positive or negative controls, respectively. The expression of all four NF-κB/Rel family genes examined was significantly upregulated in maturing DCs compared to macrophages. The strongest difference was observed for c-rel. RT-PCR determinations of c-rel, RelB, and p105 mRNAs confirmed these observations. Among the 32 NF-κB/Rel pathway genes, 14 were upregulated in mature DCs compared to macrophages. These genes were IκBα, IKK-β, NIK, ICAM-1, P-selectin, E-selectin, TNF-α, TNFR2, TN FAIP3, IL-1α, IL-1R1, IL-1 R2, IRAK, and TANK. By contrast, only mcp-1 (monocyte chemotactic protein 1) was upregulated in macrophages compared to DCs. NF-κB pathway genes upregulated in DCs compared to macrophages were constitutively expressed in monocytes then selectively downregulated during macrophage but not DC differentiation. LPS did not induce expression of most of these genes in macrophages but LPS did induce upregulation of IL-8 in mature macrophages. We conclude that NF-κB/Rel family genes, especially c-rel, are selectively expressed during differentiation of monocytes towards DCs. Moreover, this differential expression is associated both with activation of different NF-κB signal transduction pathways in DCs and macrophages and with expression of a unique subset of genes in DCs that are transcriptionally targeted by NF-κB/Rel factors. The results illustrate the ability of the NF-κB pathway to respond to differentiation stimuli by activating in a cell-specific manner unique signalling pathways and subsets of NF-κB target genes.

Original languageEnglish (US)
Pages (from-to)281-290
Number of pages10
JournalJournal of Cellular Biochemistry
Issue number2
StatePublished - Jan 1 2001


  • cDNA arrays
  • Dendritic cells
  • Macrophages
  • NF-κB

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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