Journal: Immunity

Article Title: IL-10-Dependent Crosstalk between Murine Marginal Zone B Cells, Macrophages, and CD8α + Dendritic Cells Promotes Listeria monocytogenes Infection

doi: 10.1016/j.immuni.2019.05.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: EasySepTM Mouse Pan-DC Enrichment Kit , StemCell Technologies , Cat # 19763.

Techniques: Purification, Blocking Assay, Marker, Virus, Expressing, Recombinant, Red Blood Cell Lysis, Staining, Modification, Protease Inhibitor, Reverse Transcription, Cell Isolation, Software

Journal: Immunity

Article Title: IL-10-Dependent Crosstalk between Murine Marginal Zone B Cells, Macrophages, and CD8α + Dendritic Cells Promotes Listeria monocytogenes Infection

doi: 10.1016/j.immuni.2019.05.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: EasySepTM Mouse Pan-DC Enrichment Kit , StemCell Technologies , Cat # 19763.

Techniques: Purification, Blocking Assay, Marker, Virus, Expressing, Recombinant, Red Blood Cell Lysis, Staining, Modification, Protease Inhibitor, Reverse Transcription, Cell Isolation, Software

Journal: Cell Reports

Article Title: ILC precursors differentiate into metabolically distinct ILC1-like cells during Mycobacterium tuberculosis infection

doi: 10.1016/j.celrep.2022.110715

Figure Lengend Snippet: IL-18Rα-expressing ILC differentiate into ILC1-like cells during Mtb infection (A) Representative dot plots showing the gating strategy used for ILC subsets in the lungs of Mtb -infected C57BL/6 mice (top graphs): ILC1 (dark blue), ILC3 (red), IL-18Rα − ILC2 (green), IL-18Rα + ILC (yellow), and T-bet + IL-18Rα + ILC (light blue). (B) Unsupervised t-distributed stochastic neighbor embedding (t-SNE) distribution of total lung Lin − CD90.2 + populations at steady state (left graph) and during Mtb infection (right graph). Based on the gating strategy defined in (A), ILC subsets were depicted with the same color code. (C) Expression of GATA3 (MFI), ST2 (%), and Arg1 (%) in indicated ILC subsets at day 28 post-infection in C57BL/6 mice. (D) As in (C), but for the expression of T-bet (MFI) and CD49a (%). (E) Percentages of NKp46 + (left) and Eomes + (right) cells in ILC1-like cells compared to in NK cells and ILC1 at day 28 post-infection. (F) Percentages of IFN-γ, IL-5, and IL-17A-positive cells in the indicated ILC subsets after phorbol myristate acetate (PMA)/ionomycin stimulation at day 28 post-infection in C57BL/6 mice. (G and H) Absolute numbers of ILC1-like cells at the indicated days after Mtb infection (G) and absolute numbers of IL-18Rα + ILC at the indicated days after Mtb infection (H). Before sacrifice, mice were injected with fluorescent anti-CD45.2 to distinguish vascular and parenchymal cells. ILC1-like cells and IL-18Rα + ILC have been gated on lung-resident cells. (I) Percentages of IFN-γ + cells in the indicated ILC subsets after ex vivo stimulation with IL-12+IL-18 or not at day 28 post-infection in C57BL/6 mice. (J) Experimental settings for the adoptive transfer of ST2 + IL-18Rα − and ST2 + IL-18Rα + ILC into Rag2 −/− γc −/− before Mtb infection. (K) Percentage of ILC (Lin − CD45.2 + CD90.2 + CD127 + ) in the lung at day 21 post-infection in Rag2 −/− γc −/− after adoptive transfer of ST2 + IL-18Rα − ILC (green) and ST2 + IL-18Rα + ILC (yellow). (L) As in (J), but for T-bet expression in ILCs. In (C), (D), (G) and (H), data are representative of 5 independent experiments; in (E), (F), and (I), data are representative of 2 independent experiments; and in (K) and (L), data are a pool of 2 independent experiments, with each symbol representing an individual mouse. Graphs depict data as means ±SEMs, and statistical analysis was performed using 2-way ANOVA (F and I), 1-way ANOVA (C–E and G–H) or the Mann-Whitney test (K and L). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001).

Article Snippet: Lung ILCs were enriched from lung single-cell suspensions by using the EasySepTM Mouse ILC2 Enrichment Kit (StemCell).

Techniques: Expressing, Infection, Injection, Ex Vivo, Adoptive Transfer Assay, MANN-WHITNEY

Journal: Cell Reports

Article Title: ILC precursors differentiate into metabolically distinct ILC1-like cells during Mycobacterium tuberculosis infection

doi: 10.1016/j.celrep.2022.110715

Figure Lengend Snippet: The inflammatory environment shapes the fate of IL-18Rα + ILCs (A) Representative dot plots of T-bet and IL18Rα expression after intranasal administration of PBS and IL-12+IL-18 in Rag2 −/− mice in Lin − CD45.2 + CD90.2 + NK1.1 − RORγt − cells. (B) Absolute numbers of IL-18Rα − ILC2 (green), IL-18Rα + ILC (yellow), and ILC1-like cells (blue) after cytokine (IL-12+IL-18) or control (PBS) treatment. (C) Percentage of TCF-1 in lung IL-18Rα − ILC2 after intranasal administration of PBS or IL-12+IL-18. (D) Percentages of cells expressing IL-5 or IFN-γ among the indicated ILC subsets after PMA/ionomycin stimulation. (E) Percentages of IFN-γ + cells in the indicated ILC subsets after ex vivo stimulation with IL-12+IL-18, PMA/ionomycin, or not from IL-12+IL-18-treated C57BL/6 mice. (F) Representative dot plot of T-bet and IL-18Rα expression in IL-12+IL-18-treated Stat1 +/+ versus Stat1 −/− mice. (G) Absolute numbers of ILC1-like cells (left) and IL-18Rα + ILC (right) in Stat1 +/+ versus Stat1 −/− mice treated as in (F). (H) Representative dot plots of T-bet and IL18Rα expression after IL-33, IL-12+IL-18, and IL-12+IL-18+IL-33-treated Rag2 −/− mice in Lin − CD45.2 + CD90.2 + NK1.1 − RORγt − cells. (I–K) Absolute numbers of IL-18Rα − ILC2 (I), IL-18Rα + ILC (J), and ILC1-like cells (K) in IL-33, IL-12+IL-18, or IL-12+IL-18+IL-33-treated mice. (L) Percentage of IL-5 + IFN-γ − (right), IL-5 − IFN-γ + (center), and IL-5 + IFN-γ + (left) in IL-18Rα + ILC in IL-33, IL-12+IL-18, and IL-12+IL-18+IL-33-treated mice. Each symbol represents an individual mouse. Statistical analysis was performed using Mann-Whitney (B, C, and G) and 1-way (I–L) and 2-way (D and E) ANOVA tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. Graphs depict data as means ±SEMs. Data are representative of 3 (B, D, and I–K) and 2 (C, E, G, and L) independent experiments.

Article Snippet: Lung ILCs were enriched from lung single-cell suspensions by using the EasySepTM Mouse ILC2 Enrichment Kit (StemCell).

Techniques: Expressing, Control, Ex Vivo, MANN-WHITNEY

Journal: Cell Reports

Article Title: ILC precursors differentiate into metabolically distinct ILC1-like cells during Mycobacterium tuberculosis infection

doi: 10.1016/j.celrep.2022.110715

Figure Lengend Snippet: Metabolic reprogramming toward glycolysis is associated with an ILC1-like cell differentiation (A) Representative histograms of puromycin staining in NK cells (violet), IL-18Rα − ILC2 (green), IL-18Rα + ILC (yellow), and ILC1-like cells (blue) in IL-12+IL-18-treated mice. (B) Expression of puromycin (mean fluorescence intensity [MFI]) in NK cells (violet), IL-18Rα − ILC2 (green), IL-18Rα + ILC (yellow), and ILC1-like cells (blue) in PBS versus IL-12+IL-18-treated mice. (C) Percentage of puromycin-positive cells in the indicated ILC subsets in PBS versus IL-12+IL-18-treated mice. (D) Representative histograms of puromycin staining (left) and quantification (MFI, right) in ILC1-like cells from IL-12+IL-18-treated mice after incubation with various metabolic inhibitors (Co, control; DG, 2-deoxyglycose; O, oligomycin; DGO, 2-deoxyglucose + oligomycin). (E and F) Percentages of mitochondrial dependence (E) and glycolytic capacity (F) in the indicated ILC subsets in PBS versus IL-12+IL-18-treated mice. (G) Expression of puromycin (MFI) in ILC1-like cells from Mtb -infected Rag2 −/− mice after incubation with various metabolic inhibitors. (H) Percentage of mitochondrial dependence (blue) and glycolytic capacity (red) in ILC1-like cells in Mtb -infected versus IL-12+IL-18-treated Rag2 −/− mice. (I) Expression of IFN-γ in total ILCs from Mtb -infected Rag2 −/− mice after ex vivo stimulation with IL-12+IL-18 in the presence or absence of 2-DG. (J) Mycobacterial loads at day 28 post-infection in 2-DG-treated (red) versus 30% glucose-treated (blue) versus control Rag2 −/− mice. (K) Quantification of IFN-γ in the lung supernatant of 2-DG-treated (red) versus 30% glucose-treated (blue) versus control Rag2 −/− mice at day 28 post-infection. (L) Absolute numbers of ILC1-like cells in 2-DG-treated (red) versus 30% glucose-treated (blue) versus control Rag2 −/− mice at day 28 post-infection. (M) Percentages of IFN-γ + cells in ILC1-like cells after ex vivo stimulation by PMA/ionomycin in 2-DG-treated (red) versus 30% glucose-treated (blue) versus control Rag2 −/− mice at day 28 post-infection. Each symbol represents an individual mouse, and statistical analysis was performed using 2-way (B–F) and 1-way (G–M) ANOVA and a paired t test (I). ∗ , p < 0.05; ∗∗ , p < 0.01; ∗∗∗ , p < 0.001; ∗∗∗∗ , p < 0.0001. Graphs depict data as means ±SEMs from 2 (A–M) independent experiments.

Article Snippet: Lung ILCs were enriched from lung single-cell suspensions by using the EasySepTM Mouse ILC2 Enrichment Kit (StemCell).

Techniques: Cell Differentiation, Staining, Expressing, Fluorescence, Incubation, Control, Infection, Ex Vivo

Journal: Cell Reports

Article Title: ILC precursors differentiate into metabolically distinct ILC1-like cells during Mycobacterium tuberculosis infection

doi: 10.1016/j.celrep.2022.110715

Figure Lengend Snippet: ILC1-like cells confer protection against Mtb (A) Mycobacterial loads at days 14 and 21 post-infection in Rag2 −/− γc −/− mice adoptively transferred with ST2 + ILC (black) or not (gray). (B) T-bet expression (%) at different days post-infection in transferred ILC2 in Rag2 −/− γc −/− mice. (C) Mycobacterial loads at day 21 post-infection in Rag2 −/− γc −/− mice adoptively transferred with ST2 + IL-18Rα − ILC (green) versus ST2 + IL-18Rα + ILC. (D) Schematic representation of the in vivo expansion of ILC1-like cells in Rag2 −/− mice treated with IL-12+IL-18+IL-33, cell sorting of ILC1-like cells (Lin − CD45.2 + CD90.2 + NK1.1 − ST2 − CD49a + IL-18Rα + ), and adoptive transfer in Rag2 −/− γc −/− 1 day before infection with Mtb by the intratracheal route. (E) Representative histograms of T-bet, GATA3, and RORγt expression in sorted ILC1-like cells (gray) versus ILC2 (Lin − CD45.2 + CD90.2 + NK1.1 − ST2 + cells). (F) Bacterial loads at day 21 post-infection in Rag2 −/− γc −/− mice having received (gray) or not (blue) an adoptive transfer of ILC1-like cells from IL-12+IL-18+IL-33-treated Rag2 −/− mice 1 day before Mtb infection. (G) C57BL/6 were vaccinated by intranasal administration of BCG or treated intranasally with IL-12+IL-18 or not (PBS) 60 days before Mtb infection. After 14 days post-infection, mice were euthanized. (H) Mycobacterial loads at day 14 post-infection in BCG-vaccinated (gray), IL-12+IL-18-treated (black), or control mice (white). (I) Percentages of total lung ILCs expressing T-bet. (J) Absolute numbers of NK, ILC1, IL-18Rα − ILC2, IL-18Rα + ILC, ILC3, and ILC1-like cells at day 14 post in BCG-vaccinated (gray), IL-12+IL-18-treated (black), or control mice (white). (K) Percentages of IFN-γ + cells among NK (left), ILC1 (center), and ILC1-like cells (left) in BCG-vaccinated (gray), IL-12+IL-18-treated (black), or control mice (white). after ex vivo stimulation with IL-12+IL-18. Each symbol represents an individual mouse. Statistical analysis was performed using the Mann-Whitney test (B–F) and 1-way (H–K) and 2-way (A) ANOVA tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. Graphs depict data as means ±SEMs. Data are representative of 2 (A, B, and F–K) independent experiments and a pool of 2 experiments (C).

Article Snippet: Lung ILCs were enriched from lung single-cell suspensions by using the EasySepTM Mouse ILC2 Enrichment Kit (StemCell).

Techniques: Infection, Expressing, In Vivo, FACS, Adoptive Transfer Assay, Control, Ex Vivo, MANN-WHITNEY

Journal: Cell Reports

Article Title: ILC precursors differentiate into metabolically distinct ILC1-like cells during Mycobacterium tuberculosis infection

doi: 10.1016/j.celrep.2022.110715

Figure Lengend Snippet:

Article Snippet: Lung ILCs were enriched from lung single-cell suspensions by using the EasySepTM Mouse ILC2 Enrichment Kit (StemCell).

Techniques: Software, Recombinant, Staining, Enzyme-linked Immunosorbent Assay