Project description:We analyzed the transcriptome differences of wild-type and ArhGEF1-deficient (Arhgef1-/-) type-2 conventional dendritic cells (cDC2s) in spleen. Wild-type or ArhGEF1-deficient (Arhgef1-/-) bone marrow cells were transferred into lethal irradiated CD45.1 mice. 8 weeks later, splenic cDC2s were purified and prepared for RNA-seq analysis. The gene expression profiles of CD97-, Gα13- and ArhGEF1-deficient cDC2s were highly similar, providing evidence that these molecules function in the same pathway.
Project description:We analyzed the transcriptome differences of wild-type, CD97- and Gα13-deficient (Adgre5-/- and CD11c-cre x Gna13fl/fl) type-2 conventional dendritic cells (cDC2s) in spleen. Three technical repeats of ~10^5 cells per sample from each of one mouse were included. Compared to wild-type cDC2s, CD97- and Gα13-deficient cDC2s differentially expressed many genes, but CD97- and Gα13-deficient cDC2s were almost the same. GSEA showed that Mrtf-a dependent genes were upregulated in CD97- and Gα13-deficient cDC2s, which code for cytoskeleton proteins. These data support that CD97-Gα13 signaling regulates splenic cDC2 motility by the actin cytoskeleton.
Project description:We analyzed the transcriptome differences of wild-type and IRF4-deficient (CD11c-cre x Irf4fl/fl) type-2 conventional dendritic cells (cDC2s) in spleen. Wild-type or IRF4-deficient (CD11c-cre x Irf4fl/fl) bone marrow cells were transferred into lethal irradiated CD45.1 mice. 8 weeks later, splenic cDC2s were purified and prepared for RNA-seq analysis. Three technical repeats of ~10^5 cells per sample from each of one mouse were included. CD97 was downregulated in IRF4-deficient cDC2s. GSEA showed that upregulated and downregulated genes in CD97-deficient cells were strongly enriched in IRF4-regulated genes. These data are in accord with CD97 acting within the IRF4 gene-expression network.
Project description:Our study explored the role of the ubiquitin-editing enzyme A20 (TNFAIP3) in regulating conventional dendritic cell (cDC) differentiation and its implications for anti-tumor immunity. We found that A20 stabilizes IRF8, essential for type 1 cDC (cDC1) development, by removing K48-linked polyubiquitin chain. DC-specific A20-depleted (A20cKO) mice showed a significant reduction in cDC1 levels but exhibited robust anti-tumor immunity due to superior MHC-I-mediated antigen presentation by de novo type 2 cDCs (cDC2s). These de novo cDC2s showed distinct gene expression profiles compared to wild-type cDC2s and acquired the ability to trogocytose adjacent tumor cells. NF-κB-dependent TNFα and TBK1/IRF3-mediated type I interferon secreted by cDCs in A20cKO mice played crucial roles in cDC2’s acquisition of cross-dressing characteristics, facilitating tumor-specific cytotoxic T lymphocyte responses even when cross-dressed with allo-tumor cells. This study highlights the crucial role of A20 in the development of cDC1s through IRF8 stabilization and unveils the unique anti-tumor potential of A20cKO cDC2s through their cross-dressing capabilities.
Project description:Adipose and splenic B cells are expanded with age in wild-type mice, but not Nlrp3-/- mice. This comparison is to address the transcriptional changes that are dependent upon age or Nlrp3.