Profiling dendritic cell maturation with dedicated microarrays

Abstract

Dendritic cell (DC) maturation is the process by which immature DC in the periphery differentiate into fully competent antigen-presenting cells that initiate the T cell response. However, DC respond to many distinct maturation stimuli, and different types of mature DC induce qualitatively different T cell responses. As DC maturation involves the coordinated regulation of hundreds of genes, comprehensive assessment of DC maturation status would ideally involve monitoring the expression of all of these transcripts. However, whole-genome microarrays are not well-suited for routine phenotyping of DC, as the vast majority of genes represented on such chips are not relevant to DC biology, and their cost limits their use for most laboratories. We therefore developed a DC-dedicated microarray, or "DC Chip", incorporating probes for 121 genes up-regulated during DC maturation, 93 genes down-regulated during maturation, 14 DC-specific genes, and 90 other genes with known or probable immune functions. These microarrays were used to study the kinetics of DC maturation and the differences in maturation profiles among five healthy donors after stimulation with tumor necrosis factor-alpha + polyI:C. Results obtained with the DC Chip were consistent with flow cytometry, enzyme-linked immunosorbent assay, and real-time polymerase chain reaction, as well as previously published data. Furthermore, the coordinated regulation of a cluster of genes (indoleamine dioxygenase, kynureninase, kynurenine monoxygenase, tryptophanyl tRNA synthetase, and 3-hydroxyanthranilate 3,4-dioxygenase) involved in tryptophan metabolism was observed. These data demonstrate the use of the DC Chip for monitoring the molecular processes involved in the orientation of the immune response by DC.

Identification of TTP mRNA targets in human dendritic cells reveals TTP as a critical regulator of dendritic cell maturation

“…At the molecular level, DC maturation results in dramatic changes in the gene expression profile in the cell. Most studies to date have focused on changes in transcription (Tureci et al 2003;McIlroy et al 2005). However, modulation of gene expression occurs not only at the level of transcription, but also at post-transcriptional levels such as changes in mRNA stability and translation.…”

Identification of Pancreatic Cancer-Associated Tumor Antigen from HSP-Enriched Tumor Lysate-Pulsed Human Dendritic Cells

“…Subsequent evaluation and selection by several quality criteria (see Materials and Methods) led to identification of 194 peptides derived from 64 different proteins (Supplementary Table 1, only online). Proteomic profiling of human monocyte-derived dendritic cells revealed that a number of these peptides are derived from ubiquitously expressed proteins (e.g., from house-keeping, cytoskeletal, or matrix proteins) 34,35,36,37,38,39,40,41,42,43. These include ribosomal proteins, actin and actin-related proteins, aldolase A, annexin A2, β2-micoglobulin, calmodulin, cathepsin proteins, superoxide dismutase, systatins, elonase 1, galectin 1, glycolysis-related enzymes, histones, HSP70, IL-1RA, kynureninase, macrophage migrating inhibitory factor, peroxiredoxin 6, phosphoglycerated kinase 1, S100 calcium binding proteins and vimentin.…”

Tryptophan and Indoleamine-2,3-Dioxygenase (Ido) in Pathogenesis of Immunosuppressive Clinical Conditions

“…Оказалось, что IFNγ в ДК прак- 2017, Vol. 19, 3 2017 тически одновременно индуцирует экспрессию этой цепочки ферментов, которые отвечают в виде скоординированного кластера генов [41]. Более того, IFNγ способен индуцировать экс-прессию данной цепочки генов в CD8-ДК у мы-шей при отсутствии индукции в них IDO, когда эти ДК поглощают Кин, продуцированными дру-гими ДК, в которых Кин образовывался из Три под влиянием IDO.…”