Transcript profiling of human dendritic cells maturation‐induced under defined culture conditions: comparison of the effects of tumour necrosis factor alpha, soluble CD40 ligand trimer and interferon gamma

Abstract

Summary. Using cDNA arrays, we characterized patterns of gene expression in populations of human dendritic cells (DCs) produced for clinical use. Culture and maturation induction of myeloid adherent cells under serum-free conditions yielded DCs with phenotypes similar to those described in serum-based systems. Analysis of gene expression in DCs treated with tumour necrosis factor alpha, soluble CD40L trimer or interferon gamma, however, showed specific patterns for each factor examined. Our studies document the expression of several transcripts that have not hitherto been described in DCs and/or differentially regulated according to the differentiation state of the DCs, and suggest important functional differences among the DC populations examined. In addition, DC maturation directs changes in the levels of mRNA specific for transcriptional regulators that effect the production of cytokines (e.g. BCL-6, c-rel). Other changes observed, including alteration in the gene expression profile of adhesion molecules and chemokine receptors such as CD44H, CD 49B, Rantes R, CXCR5 and CD37, suggest differences in trafficking potential between the populations studied. This broad-based description of DC populations, produced under serum-free conditions, has enabled us to better define intermediate stages of DC maturation as well as the differentiation-inducing effects of cytokines on these cells.

Transcriptional profiles during the differentiation and maturation of monocyte-derived dendritic cells, analyzed using focused microarrays

“…As in previous reports [11,12], two of the three GADD45 genes probed in our microarrays were found to be strongly up-regulated upon stimulation with LPS and CD40L.…”

“…Remarkably, the transcript level for IL-1B was ~9-fold higher in LPS-stimulated DC than in iDC-3d. In some reports, the gene expressions of IL-1B and IL-6 were respectively up-regulated in response to CD40L and TNF-α [12,13], suggesting that mature DC may be an important source of pro-inflammatory cytokines. As in previous studies [13], we failed to detect the transcription of the gene encoding the alpha subunit of IL-12 (IL-12A, p35) for any of the stimulus groups, although the gene coding for the beta subunit of IL-12 (IL-12B, p40) was found to be induced selectively in response to CD40L.…”

“…A growing body of evidence shows that DC respond differently to different stimuli, and that depending on the stimuli, the T cells that come into contact with the DC will acquire diverse effector functions [8,9]. Thus, an increasing number of researchers are using high-density microarray techniques to try to clarify the underlying changes and differences in gene expression via separate or comparative analyses of DC matured with different stimuli [10][11][12][13]. Many highly parallel gene expression analyses of immature DC and monocytes have also been performed [14,15].…”

Comparative Analysis of Signal Transduction by CD40 and the Epstein-Barr Virus Oncoprotein LMP1 In Vivo

“…The expression of LMP1CD40 in the B lymphocytes of transgenic mice led to the upregulation of the cell surface activation markers CD69, CD80, and CD86, which are also upregulated in response to CD40 activation. LMP1CD40 suppressed BCL6 mRNA and protein levels, similar to CD40 activation (1,8,24). Finally, the suppression of GC formation which was caused by LMP1CD40 expression has also been observed in mice by the administration of CD40 agonists over a period of 1 week (6).…”

A Regulatory Role for CD37 in T Cell Proliferation

“…In humans, CD37 expression is restricted to B and T lymphocytes, monocytes, macrophages, neutrophils, dendritic cells (DC) 3 , and certain leukocyte-derived malignant cells (21,22). CD37 is not expressed by NK cells, platelets, or erythrocytes.…”