Notch and TLR signaling coordinate monocyte cell fate and inflammation
- Vascular Medicine Research, Hannover Medical School, Germany
- Department of Nephrology and Hypertension, Hannover Medical School, Germany
- Institut für Kardiovaskuläre Physiologie, Fachbereich Medizin der Goethe-Universität Frankfurt am Main, Germany
- Department of Plastic, Aesthetic, Hand and Reconstructive Surgery, Hannover Medical School, Germany
- Institute of Medical Microbiology and Hospital Epidemiology, Hannover Medical School, Germany
- Mucosal Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, Germany
- Center for Cardiology, Cardiology I, University Medical Center of the Johannes Gutenberg-University Mainz, Germany
- Center for Thrombosis and Hemostasis, University Medical Center of the Johannes Gutenberg-University Mainz, Germany
- German Center for Cardiovascular Research (DZHK), Partner Site Rhine Main, Germany
- Department of Medical Microbiology and Hygiene, Medical Center of the Johannes Gutenberg-University of Mainz, Germany
- Institute for Experimental Infection Research, TWINCORE, Centre for Experimental and Clinical Infection Research, a joint venture between the Helmholtz Centre for Infection Research Braunschweig and the Hannover Medical School, Germany
- Cluster of Excellence-Resolving Infection Susceptibility (RESIST), Hanover Medical School, Germany
- Department of Surgery, Klinikum rechts der Isar, Technical University Munich, Germany
Figures
Figure 1 with 1 supplement
Inflammatory conditions enhance monocyte conversion in vitro.
(A–F) Monocyte conversion in the presence of DLL1 and TLR agonists in vitro: (A) Representative flow cytometry plot, (B) relative frequency of Ly6Clo monocyte-like cells in live CD11b+GFP+ cells …
Figure 1—figure supplement 1
Strategy of monocyte isolation from mouse bone marrow.
Figure 2 with 1 supplement
Inflammatory conditions enhance monocyte conversion in vivo.
(A–E) Adoptive transfer and flow cytometry analysis of BM CD45.2+ Ly6Chi monocytes in control or R848 injected CD45.1+ congenic recipients: (A) Experimental setup is depicted; (B) Flow cytometry …
Figure 2—figure supplement 1
Inflammatory conditions enhance monocyte conversion in vivo.
(A, B) Full gating path of adoptively transferred donor CD45.2+ monocytes recovered from CD45.1+ recipients corresponding to Figure 2B and D respectively.
Figure 3 with 3 supplements
Acute inflammation triggers altered myeloid cell response in N2ΔMy mice.
(A) Experimental set-up for IMQ treatment and analysis of mice. (B, C) Gating strategy for t-SNE analysis and definition of cell subsets based on expression of surface markers are shown. t-SNE was …
Figure 3—figure supplement 1
IMQ treatment induces systemic inflammation in mice.
(A) Body weight (left) and ear thickness (right) of IMQ-treated wt or N2ΔMy mice (data are from three experiments, n = 11/13). (B) Spleen weight and cell number of IMQ-treated wt or N2ΔMy mice (data …
Figure 3—figure supplement 2
Identification of myeloid cell subsets in IMQ-treated mice.
(A) Gating strategy for definition and quantification of myeloid subsets from live CD45+Lin-CD11b+GFP+ cells by flow cytometry. Defined populations are color-coded and used for subsequent …
Figure 3—figure supplement 3
Flow cytometry analysis of myeloid cells in IMQ-driven inflammation.
(A) Absolute frequency of different myeloid subpopulations in PB. (B) Absolute frequency of different myeloid subpopulations normalized per mg tissue (top) or per spleen (bottom). (C) Relative (top) …
Figure 4 with 1 supplement
Enhanced macrophage gene expression signatures in monocytes and altered inflammatory response in N2ΔMy mice.
(A, B) Hierarchical clustering of 600 ANOVA-selected DEGs (A) and PCA of PB monocyte subsets (B) after IMQ treatment (n = 4) is shown (Variance filtering 0.295, ANOVA followed by the B-H correction …
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Figure 4—source data 1
List of 600 DEGs for hierarchical clustering and PCA (Figure 4A and B) from Ly6Chi and Ly6Clo subpopulations isolated from sham-or IMQ(Aldara)-treated wt or N2ΔMy mice.
- https://cdn.elifesciences.org/articles/57007/elife-57007-fig4-data1-v2.xlsx
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Figure 4—source data 2
List of 373 DEGs between Ly6Clo cells isolated from IMQ(Aldara)-treated wt or N2ΔMy mice and used for the analysis in Figure 4C and D, Figure 5A and Supplementary file 2–5.
- https://cdn.elifesciences.org/articles/57007/elife-57007-fig4-data2-v2.xlsx
Figure 4—figure supplement 1
Gating strategy for cell sorting, IPA and cytokine and chemokine analysis in IMQ-treated mice.
(A) Sorting strategy of monocyte subsets for RNA-seq analysis. Lin-CD11b+GFP+CD115+Ly6ChiCD43-MHC-II- (Ly6Chi) and Lin-CD11b+GFP+CD115+Ly6Clo/-MHC-II- (Ly6Clo) monocytes were sorted from naive or …
Figure 5 with 2 supplements
Notch2-deficient Ly6Clo cells show enhanced macrophage maturation during acute inflammation.
(A) GSEA based on 373 DEGs between IMQ-treated wt and N2ΔMy Ly6Clo subsets in PB. Red – positive-, and blue - negative enrichment in corresponding color-coded wt or N2ΔMy cells. Size of the circle …
Figure 5—figure supplement 1
Characterization of F4/80hi monocytes in IMQ-treated mice.
(A) Experimental set-up for IMQ treatment, adoptive transfer and analysis of mice is depicted. (B) t-SNE analysis of donor CD45.2+CD11b+GFP+ cells extracted from Spl of IMQ-treated mice. Overlay of …
Figure 5—figure supplement 2
Expression of Notch2 and Notch-regulated gene in monocytes of IMQ-treated mice.
(A, B) Bar graph showing mean + SEM of Notch2 (A) or Hes1 (B) sequence reads from IMQ-treated wt and N2ΔMy PB monocyte subsets. (C) Representative flow cytometry plots showing expression of Notch2 …
Additional files
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Supplementary file 1
Surface phenotype signatures for identification of distinct myeloid populations in vivo.
Lin: CD3, CD45R/B220, CD19, NK1.1, Ly6G, Ter119.
- https://cdn.elifesciences.org/articles/57007/elife-57007-supp1-v2.doc
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Supplementary file 2
IPA of top five immunological diseases enriched in Ly6Clo cells from IMQ-treated N2ΔMy mice.
- https://cdn.elifesciences.org/articles/57007/elife-57007-supp2-v2.doc
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Supplementary file 3
Top 20 gene sets involved in GO biological processes enriched in Ly6Clo cells from IMQ-treated N2ΔMy mice.
- https://cdn.elifesciences.org/articles/57007/elife-57007-supp3-v2.doc
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Supplementary file 4
Parameters and the results of GSEA performed on 373 DEGs for Figure 5A.
- https://cdn.elifesciences.org/articles/57007/elife-57007-supp4-v2.xlsx
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Supplementary file 5
List of the genes enriched in Lyve1hiMHC-IIlo MF gene set from Figure 5A.
- https://cdn.elifesciences.org/articles/57007/elife-57007-supp5-v2.xlsx
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Supplementary file 6
List of antibodies and fluorescence dyes for flow cytometry used in the study.
- https://cdn.elifesciences.org/articles/57007/elife-57007-supp6-v2.doc
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Transparent reporting form
- https://cdn.elifesciences.org/articles/57007/elife-57007-transrepform-v2.pdf
