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Identification of a human cDC2 homeostatic maturation program by scRNA-seq

ABSTRACT: To explore the homeostatic maturation program of human cDCs, we exploited the nature of the spleen where DC maturation leads to migration from the red pulp and marginal zone to the white pulp of the spleen. Therefore, all steps of DC maturation are present and can be studied in the same organ. We first used single-cell RNA-seq (scRNA-seq) to profile lineage- (lin-) CD11c+ HLA-DR+ cells from spleens of 3 healthy human donors and identified a homeostatic maturation of the cDC2 subtype with 3 stages (cDC2, DC2pre-hm, DC2hm).

ORGANISM(S): Homo sapiens

PROVIDER: GSE267253 | GEO | 2026/02/23

REPOSITORIES: GEO

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Project description:XCR1+ dendritic cells (DC) have been shown to excel in antigen cross-presentation for the activation of naÃ¯ve CD8 T cells. This property was reported to be associated to the subset of the XCR1+ DC expressing IL-12b upon ex vivo stimulation for 24 h with a mixture of CpG, IFN-Î³, and GM-CSF (Lin ML et al. Proc Natl Acad Sci USA. 2008. PMID: 18272486). DC found in the steady-state non-lymphoid tissues undergo an homeostatic, tolerogenic, maturation and migrate to the draining lymph nodes to interact with naive autoreactive T cells and induction their peripheral tolerance. In contrast, spleen DC are thought to exist solely in an immature state. The aim of this study was to re-examine heterogeneity within steady state spleen XCR1+ DC, in particular examining whether this population encompass a fraction of mature DCs as assessed through their expression of CCR7 and/or the Il12b gene. Indeed, we show that a small fraction of XCR1+ spleen DC constitutively mature into two distinct but likely successive activation stages characterized as CCR7+ and CCR7+Il12b+ respectively, and correlated with increasing ability to cross-present antigen to naÃ¯ve CD8 T cells. Transcriptomic analysis of the subsets of XCR1+ DC found in steady state spleen unexpectedly showed that their homeostatic maturation was unexpectedly associated with up-regulated of many genes thought to drive pro-inflammatory T-cell responses and previously found to be commonly induced upon maturation of distinct DC subsets in response to stimulation by various microbial-type stimuli (Vu Manh TP et al. Eur J Immunol. 2013. PMID: 23553052). Thus, our results reveal that spleen XCR1+ DC undergo constitutive maturation and emphasize the common mechanisms operating upon homeostatic, tolerogenic, DC maturation versus microbial-type stimuli-induced, immunogenic, DC maturation. DC were isolated from the spleen of untreated Il12b-EYFP reporter mice (Reinhardt RL et al. J Immunol. 2006. PMID:16849470) mice as previously described (Robbins SH et al. Genome Biol. 2008. PMID: 18218067; Baranek T et al. Cell Host Microbe. 2012. PMID: 23084923). DC subsets were sorted by flow cytometry according to the marker combinations described in the âcharacteristics: phenotypeâ field for each sample.

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Identification of a human cDC2 homeostatic maturation program by scRNA-seq

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