ScATAC-seq of innate immune cells from mice treated with the adjuvant 3M052
Ontology highlight
ABSTRACT: Purpose: to explore the epigenomic landscape of innate immune cells stimulated with a novel adjuvant, 3M052 Methods: Mice were injected with 3M052. Draining lymph nodes were negatively selected for CD19+ and CD3+, then flow sorted into four populations: Dendritic cells (DCs), Double positive cells (DP, CD11b+BST1+), Ly6c+ cells (Ly6c), and plasmacytoid dendritic cells (pDCs). Lymph nodes were harvested at baseline (D0), 24 hours post-treatment (D1) or 28 days post-treatment (D28).
Project description:Purpose: To understand the innate immune response to an adjuvant, 3M052, and yellow fever vaccine, YFV Methods: Draining lymph nodes were negatively selected for CD19+ and CD3+, then flow sorted into four populations: Dendritic cells (DCs), Double positive cells (DP, CD11b+BST1+), Ly6c+ cells (Ly6c), and plasmacytoid dendritic cells (pDCs). Lymph nodes were harvested at baseline (D0), 24 hours post-treatment (D1) or 28 days post-treatment (D28). Results: TBD Conclusions: TBD
Project description:Gamma-delta (gd) T cells from pooled mouse lymph nodes and spleens were isolated and FACS-sorted for Vg1+Ly6C-, Vg1+Ly6C+, Vg4+Ly6C- and Vg4+Ly6C+ subsets of bulk CD27+ gdT cells.
Project description:Topical (epicutaneous, e.c.) application of the adjuvant CpG ODN during immunization leads to a robust immune response compared to when subcutaneous (s.c.) administration. Dendritic cells are hematopoietically derived cells that are important in cross-presenting to and activating CD8 T cells. Dermal dendritic cells are one of the two major dendritic cell subsets found in the skin which mobilize from the skin to draining lymph nodes to present to T cells upon activation. Dermal dendritic cells are found in skin draining lymph nodes around 24 hours post immunization. To determine how the immune system respond differently between e.c. versus s.c. administration of CpG ODN, we evaluated changes in the skin draining lymph node environment upon the two routes of adjuvant application. Expression chemokines and chemokine receptors were assessed with real-time qPCR. To determine the changes in the skin draining lymph node environment (cytokine and cytokine receptor levels) upon immunization via real time RT-PCR.
Project description:Spleen and lymph node dendritic cells have a differential capacity do induce and retain iTreg cells. Therefore we performed a comparative analysis of the dendritic cells derived from these two compartments to identify the responsible genes CD11c+ conventional dendritic cells were facs sorted from spleens and lymph nodes of BALB/c animals for RNA extraction and hybridization on Affymetreix microarray
Project description:Microarray was performed on PBMC with an inflammatory dedicated slide of 340 genes, and target samples labelled with Cy3 for day 0 (reference) and with Cy5 for the other days. Changes in expression levels were evaluated by the ratio Dx/D0. According to time evolution, a two-dimensional clustering was performed. Keywords: time evolution of sepsis in patients at D1, D7 and D28 post-inclusion
Project description:Among the childhood diseases, B-cell acute lymphocytic leukemia (B-ALL) is the most frequent type of cancer. In spite of recent advances concerning disease treatment, cytotoxic chemotherapy remains as the first line of treatment in several countries, and the modifications induced by such drugs in the organism remains poorly understood. In this context, the present study provided a comparative high-throughput proteomic analysis of the cumulative changes induced by chemotherapeutic drugs used in the induction phase of B-LLA treatment in both peripheral blood (PB) and bone marrow compartment (BM) samples. To reach this goal, PB and BM plasma samples were comparatively analyzed by using label-free proteomics at two endpoints: at diagnosis (D0) and the end of the cumulative induction phase treatment (D28). The resulting differentially expressed proteins were explored by bioinformatics approaches aiming to identify the main gene ontology processes, pathways and transcription factors altered by chemotherapy, as well to understand B-LLA biology in each compartment at D0. At D0, PB was characterized as a pro-inflammatory environment, with the involvement of several downregulated coagulation proteins as KNG, plasmin and plasminogen. D28 was characterized predominantly by immune response-related processes, and the super expression of the transcription factor IRF3 and transthyretin. RUNX1 was pointed out as a common transcription factor found in both D0 and D28. Considering that most of these proteins were not described in B-ALL literature, these findings added to understanding disease biology at diagnosis, and highlighted some important proteins and processes that may contribute to our understanding about the mechanisms concerning the impact of chemotherapy in disease resolution.
Project description:To investigate tissue of origin effects on migratory dendritic cell (DC) gene expression in shared lymph nodes we performed bulk RNAseq of DCs from each tissue (liver, pancreas, duodenum). We then performed single cell sequencing of the DCs within the draining lymph nodes.