Relationships Among Murine CD11chigh Dendritic Cell Subsets as Revealed by Baseline Gene Expression Patterns

Abstract

The functional relationships and properties of different subtypes of dendritic cells (DC) remain largely undefined. To better characterize these cells, we used global gene analysis to determine gene expression patterns among murine CD11chigh DC subsets. CD4+, CD8α+, and CD8α− CD4− (double negative (DN)) DC were purified from spleens of normal C57/BL6 mice and analyzed using Affymetrix microarrays. The CD4+ and CD8α+ DC subsets showed distinct basal expression profiles differing by >200 individual genes. These included known DC subset markers as well as previously unrecognized, differentially expressed CD Ags such as CD1d, CD5, CD22, and CD72. Flow cytometric analysis confirmed differential expression in nine of nine cases, thereby validating the microarray analysis. Interestingly, the microarray expression profiles for DN cells strongly resembled those of CD4+ DC, differing from them by <25 genes. This suggests that CD4+ and DN DC are closely related phylogenetically, whereas CD8α+ DC represent a more distant lineage, supporting the historical distinction between CD8α+ and CD8α− DC. However, staining patterns revealed that in contrast to CD4+ DC, the DN subset is heterogeneous and comprises at least two subpopulations. Gene Ontology and literature mining analyses of genes expressed differentially among DC subsets indicated strong associations with immune response parameters as well as cell differentiation and signaling. Such associations offer clues to possible unique functions of the CD11chigh DC subsets that to date have been difficult to define as rigid distinctions.

The TNF receptor and Ig superfamily members form an integrated signaling circuit controlling dendritic cell homeostasis

“…However, these two transgenic experimental conditions do not reflect physiological conditions, but may approximate inflammatory conditions, representing conditions where the equilibrium between the different mechanisms regulating steady state lymphoid tissue DC homeostasis is perturbed. Interestingly, a gene chip analysis on splenic DC subsets showed LTβ mRNA was the highest expressed amongst all mRNA between DC subsets [24]. This study also indicated that CD8α− DC subsets that are regulated via LTβR signaling express 5-to 6-fold higher LTβ mRNA levels compared to CD8α+ DC subset.…”

Anatomic location defines antigen presentation by dendritic cells to T cells in response to intravenous soluble antigens

“…A number of experimental models, most performed with splenic DC, suggest that each subset serves a unique role in the immune system [2,[6][7][8][9][10][11][12][13][14][15][16]. Moreover, CD8a + and CD8a -splenic DC have different gene-expression profiles, a finding that strongly supports a functional difference between these two subsets [17].…”

Transcription factor networks in dendritic cell development

“…In addition to the CD8-alpha positive DC, two other populations of conventional DCs (cDC) are defined by the presence or absence of the cell-surface molecule CD4 [8]. These two populations show a high degree of similarity with respect to function, development, and gene expression, and for simplicity will be grouped together and further referred to as CD4 + cDCs in this review [9]. A fourth population of plasmacytoid DCs (pDC) is defined by the expression of several distinguishing markers including B220, Siglec-H, and Bst2 [10][11][12].…”