scRNA-seq analysis of dendritic cells in skin draining lymph nodes of CLEC4C-DTR mice treated with FITC hapten

Skin draining lymph node dendritic cell populations are known to be diverse and distinguishable via their migratory or resident properties as well as by lineage. We analyzed these populations at steady state and during the sensitization phase of the Th2-driven FITC-hapten contact hypersensitivity (CHS) model. We also analyzed the dendritic cells populations of CLEC4C-DTR+ mice treated with diptheria toxin, which demonstrate exacerbated pathology in the FITC-CHS model. We demonstrate the diversity of dendritic cell populations at steady state and during type 2 inflammation, and surprisingly, reveal a skewing of the DC populations in DT treated CLEC4C-DTR+ mice, with an enrichment of DC2 migratory populations and depletion of CD326+ migratory cells in addition to pDC. Isolated mouse dendritic cells were subjected to droplet-based 3’ end massively parallel single-cell RNA sequencing using Chromium Single Cell 3’ Reagent Kits as per manufacturer’s instructions (10x Genomics). The libraries were sequenced using Illumina HiSeq4000 sequencers. CD11c+ MHCII+, Siglec H+ CD11c+ from the inguinal and axillary skin draing lymph nodes of DT-treated CLEC4C-DTR+ and CLEC4C-DTR- mice treated with epicutaneous FITC hapten were isolated after 48h of treatment. SiglecH+GFP–, SiglecH+GFPlow/int and SiglecH–GFPhigh cells from inguinal and axillary skin DLN of untreated Zbtb46GFP x CLEC4C-DTR+ were also isolated for comparison. Cells were submited for scRNAseq via the 10x Genomics platform.