Project description:We investigated the transcriptional profile of B cells isolated from peripheral lymph nodes of CCR7-/-C57BL/6 and C57BL/6 mice respectively. B cells were sorted as follows: CD19+, CD3-, CD11c-, CD11b-, NK1.1-, GR-1-. Very few genes were differentially regulated in CCR7-/- B cells, however the expression of a number of genes associated with B-cell activation was increased in CCR7-/- B cells compared to WT B cells. Our data suggest a role of CCR7 signaling in B-cell activation processes.
Project description:We investigated the transcriptional profile of B cells isolated from peripheral lymph nodes of CCR7-/-C57BL/6 and C57BL/6 mice respectively. B cells were sorted as follows: CD19+, CD3-, CD11c-, CD11b-, NK1.1-, GR-1-. Very few genes were differentially regulated in CCR7-/- B cells, however the expression of a number of genes associated with B-cell activation was increased in CCR7-/- B cells compared to WT B cells. Our data suggest a role of CCR7 signaling in B-cell activation processes. 12 purified RNA samples (6 samples of each genotype), originated from 12 individually facs-sorted cell samples, were used for the microarray study. 3 RNA samples per genotype were pooled, giving rise to 2 wildtype- and 2 Ccr7 knock-out pools. In each of the 2 dual-color microarray hybridizations, samples from 1 wildtype and 1 Ccr7 knock-out pool were cohybridized. Microarray hybridizations 1 and 2 were performed in a dye-swap approach.
Project description:To better understand how Tritrichomonas arnold colonization impacts the reovirus-mediated proinflammatory response in mesenteric lymph nodes, we examined the transcriptional profile of mesenteric lymph nodes For RNA-sequencing single cell suspension of mesenteric lymph nodes were lysed in RLT buffer (Qiagen) and RNA was isolated
Project description:We report the transcriptional profiles of tumor-specific CD8+ T cells isolated from tumor draining lymph nodes (TDLN) and tumor following tumor vaccination in C57BL/6 mice. In addition, we included wild type (WT) and TGF-b receptor conditional knockout (Tgfbr2-/-) CD8+ T cells.
Project description:Antigen presenting dendritic cells (DCs) and monocytes capture and transport antigens from barrier tissues for presentation to antigen-specific T cells in the draining-lymph nodes (LNs). While DCs enter LNs through afferent lymphatics in a CCR7-dependent manner, how exactly antigen-carrying monocytes reach LNs is less clear since monocytes do not express CCR7 and can also enter LNs via the bloodstream. In steady state, and following injection of several PAMPs, scRNA-seq data on LN mononuclear phagocytes identified LN resident versus migratory type 1 and type 2 conventional (c)DCs despite downregulation of DC subset-defining transcripts, such as Xcr1, Clec9a, H2-Ab1, Sirpa, and Clec10a on migratory cDCs. Migratory cDCs gained expression of transcripts controlling cellular migration such as Ccr7, Ccl17, Ccl22, and Ccl5, while migratory monocytes expressed Ccr5 without Ccr7. Using two tracking methods and a gating strategy that clearly distinguishes migratory CD88hiCD26lo monocytes from CD88-CD26hi cDCs, we found that both captured antigens in the lung and migrated to lung-draining LNs. Using global and mixed-chimeric Ccl5-, Ccr2-, Ccr5-, Ccr7-, and Batf3-deficient mice, we found that CCR5+ monocytes follow CCL5-secreting migratory cDCs to reach the draining LN via lymphatic vessels. In a model of asthma, such recruited monocytes regulated the induction of type 2 immunity. Overall, our data suggest that CCL5-secreting migratory cDCs lay down the chemokine trail for CCR5+ antigen-presenting monocytes to reach draining lymph nodes and regulate adaptive immunity.
Project description:Single-cell RNA sequencing (scRNA-seq) was utilized to unbiasedly dissect the cellular heterogeneity of wt vs. Rbx1-deficient CD4+YFP+ Treg cells from the peripheral lymph nodes of inflammation-free female Foxp3cre/wt and Foxp3cre/wt;Rbx1fl/fl mice.
Project description:This file contains gene microarray data from FACS purified mouse high endothelial cells and capillary endothelial cells from peripheral lymph nodes, mesenteric lymph nodes, and Peyer’s patches. The data will allow for better understanding of the specialization of high endothelial venules (HEV) and their role in lymphocyte recruitment from the blood; the tissue-specific differentiation of lymphoid tissue vasculature; and the specialized features of capillary vs. post-capillary endothelium, including differences in signaling pathways, adhesive properties and mechanisms of hemostasis.
