Project description:Using mouse lung resident conventional CD11b+ dendritic cells (CD11b+ cDCs) in the context of house-dust mite (HDM)-driven allergic airway sensitization as a model, we aimed here to identify transcriptional events regulating the pro-Th2 activity of cDCs. We used microarray analyses to identify genes differentially expressed by lung CD11b+ conventional dendritic cells in response to house dust mite allergens in wild-type and Irf3-deficient mice
Project description:Conventional dendritic cells (cDCs) in the lung that drive effector T cell differentiation, and initiate allergic responses upon inhalation of allergens. However, since CD11b+ cDCs are heterogeneous, the specific function of each subpopulation has been elusive. To identify subpopulations of CD11b+ cDCs, we performed single cell RNA-sequencing (scRNAS-Seq) of total lung CD11b+ cDCs following inhalation of house dust extracts, and detected multiple clusters based on their differential transcriptomes. The data will be applicable for studies of cDC function in induction of T cell differentiation as well as the maturation pathway of cDCs.
Project description:Here, we use a microfluidics-based approach to prepare single-cell RNA-Seq libraries from 164 primary human conventional dendritic cells (cDCs) as well as cord blood (452) and blood (341) pre-cDCs. We examined heterogeneity between individual cells to document potential subpopulations within human cDCs and pre-cDCs.
Project description:Conventional dendritic cells (cDCs) in the lung drive effector T cell differentiation, and initiate allergic responses upon inhalation of allergens. However, CD11b+ cDCs are composed Ly-6C+ and Ly-6C– subpopulations, and the specific function of each subpopulation has been elusive. Since Ly-6C+ CD11b+ cDCs lost the surface display of Ly-6C upon ex vivo culture, we hypothesized that Ly-6C+ cDCs are precursors of Ly-6C– cDCs. To determine whether 2 CD11b+ cDC subpopulations become the same cell types after maturation, we compared their transcriptomes after ex vivo cultue. As controls, we also compared transcriptomes of freshly isolated Ly-6C+ and Ly-6C– cDCs. Their transcriptomes were also compared with another CD103+ cDCs and monocytes.
Project description:To characterize differences between BALB/c splenic CD11cintB220+Gr1+ PDCs (plasmacytoid dendritic cells), CD11cintB220+CD49b+ IKDCs (interferon producing killer-dendritic cells), and CD11chighB220- cDCs (conventional dendritic cells), we performed gene expression profile analysis using Affymetrix chips. We FACS-sorted BALB/c spleen DC subpopulations. Comparison of differentially expressed genes between IKDCs and cDCs vividly revealed selective expression of multiple NK-related genes in IKDCs . These included granzymes A, B, K and M, perforin, Fas ligand, and NK receptors such as NKG2A, NKG2D, Ly49 family genes, NKR-P1, NKG7, NKp46 and Mafa (KLRG1). No NK-related genes were highly expressed in the PDCs. Experiment Overall Design: We prepared two biological samples separately for each DC population, and analyzed the expression profiles by comparing to those of cDCs (control sample).