Fig 1: Therapeutic effects of CB2 engagement depend on CB2 and IL-13 expression on ILC2s(A) Cohorts of CB2−/− mice were either adoptively transferred with WT ILC2s isolated from C57BL/6J mice or left untreated. Mice were then treated with CB2 agonist (1 mg/kg/mouse) or vehicle control every 4 days and fed an HFD for 14 weeks. n = 6.(B and C) Mice weights (B) and fasting blood glucose level (C) were measured every 2 weeks for 14 weeks.(D) Glucose tolerance test were measured after 14 weeks of treatment.(E) Cohorts of CB2−/− mice were adoptively transferred with ILC2s from either WT, IL-5−/−, or IL-13−/− mice and then treated with CB2 agonist (1 mg/kg/mouse) or vehicle control and fed an HFD for 14 weeks. n = 6.(F and G) Mice weights (F) and fasting blood glucose level (G) were measured every 2 weeks for 14 weeks.(H) Glucose tolerance test was performed after 14 weeks of treatment.(I and J) VAT weight (I) was measured and the number of ILC2s per gram of VAT (J) was quantified 14 weeks after adoptive transfer.Data are representative of 3 independent experiments, and bar graphs are shown as mean ± SEM. Statistical analyses involved one-way ANOVA (B, C, F, and G), two-way ANOVA (D and H), and two-tailed Student’s t test (I and J). Mouse image used with permission from Servier Medical Art.
Fig 2: CB2 engagement enhances activity and proliferation of activated ILC2s(A) C57BL/6J mice were assigned to receive a normal chow diet (NCD) for 14 weeks, followed by intraperitoneal (i.p.) administration of recombinant mouse (rm)IL-33 (0.5 μg in 50 μL) or PBS for 3 days. On the 4th day, the mice were euthanized, and visceral adipose tissue (VAT) was collected. n = 6.(B) Gating strategy of Lin−CD45+IL-7R+ST2+ILC2s.(C) CB2 expression in naive and IL-33-activated murine VAT-derived ILC2s versus the vehicle control. CB2 expression is quantified as mean fluorescence intensity (MFI).(D–F) Naive and activated VAT ILC2s were cultured with rmIL-2 and rmIL-7 and stimulated with CB2 agonist (10 μg/mL) or vehicle control (2% dimethyl sulfoxide) for 48 h. Levels of IL-5, IL-6, and IL-13 in culture supernatants were assessed by LEGENDplex (D). Intranuclear expression of Ki67 (E) and GATA-3 (F) were quantified as MFI.Data are representative of 3 independent experiments, and bar graphs are shown as mean ± SEM. Statistical analyses involved one-way ANOVA (C) and two-tailed Student’s t test (D–F); n.s.: p > 0.05. Mouse image used with permission from Servier Medical Art.
Fig 3: CB2 engagement enhances human ILC2 activity(A) Human peripheral blood ILC2s were freshly isolated and cultured with 10 ng/mL of recombinant human (rh)IL-2, rhIL-7, and rhIL-33 for 48 h.(B) The gating strategy for Lin−CD45+CD127+CRTH2+ human ILC2s.(C) Flow cytometry analysis of CB2 expression on naive and IL-33-activated ILC2s, quantified as MFI with SEM.(D) Naive and activated ILC2s were cultured with 10 ng/mL of rhIL-2, rhIL-7, and either CB2 agonist or vehicle control for 48 h. Levels of IL-5, IL-6, and IL-13 in the culture supernatants were quantified by LEGENDplex.Data are representative of five individual blood donors, presented as mean ± SEM. Statistical analysis involved one-way ANOVA (C) or two-tailed Student’s t test (D); n.s.: p > 0.05. Human image used with permission from Servier Medical Art.
Supplier Page from BioLegend for LEGENDplex™ MU Th2 Panel (6-plex) w/ FP V03