Journal: The Journal of Experimental Medicine

Article Title: CCL5-producing migratory dendritic cells guide CCR5 + monocytes into the draining lymph nodes

doi: 10.1084/jem.20222129

Figure Lengend Snippet: Alum OVA model and CpG immunotherapy. (A) 24 h after instillation of 20 μg of CpG and 3 μg OVA-AF488, LLNs were harvested and analyzed. Histograms illustrate expression of costimulatory molecules (CD40, CD80, and CD86) on Ag + monocytes and migratory DCs in the LLN. (B) Alum OVA model. Flow plots illustrate gating strategy for eosinophils in the BAL. Graph bar, OVA-specific IgE production after alum OVA ± pretreatment with CpG and OVA. **P < 0.01, ***P < 0.001, mean ± SEM, one-way ordinary ANOVA, with post hoc Tukey’s multiple comparison test (B).

Article Snippet: Serum was extracted to measure circulating OVA-specific IgE (RnD systems).

Techniques: Expressing, Comparison

Journal: The Journal of Experimental Medicine

Article Title: CCL5-producing migratory dendritic cells guide CCR5 + monocytes into the draining lymph nodes

doi: 10.1084/jem.20222129

Figure Lengend Snippet: Monocyte migration to draining L Ns depends on CCR5 expression. (A) 24 h after instillation of 20 μg of CpG and 3 μg OVA-AF488, LLNs were harvested and analyzed in WT and Ccr5 −/− mice. Flow plots, myeloid cells plotted as Ly6C vs. OVA + AF488 + to gate on Ag + monocytes and DCs with scatter plot showing the total number of Ag + monocytes in WT and Ccr5 −/− mice. Data combine two independent experiments; n = 4–5 per group. (B) WT, Ccr5 −/− , and Ccr2 −/− mice were analyzed for extravascular monocyte migration into the lungs after immunization. Data represent two independent experiments. Each dot represents one mouse. (C) WT mice were treated with 20 μg Maraviroc (CCR5 inhibitor) 4 h prior to i.n. delivery with 20 μg of CpG and 3 μg OVA-AF488. Scatter plot displays the number of Ag + monocytes in the LLN. Data combine two independent experiments with four to five mice per group. (D) Scatter plot illustrates the number of Ag + monocytes in the LLNs of Ccr2 −/− :WT and Ccr2 −/− :Ccr5 −/− BM chimeric mice. Data combine three independent experiments with four to five mice per group. (E) WT, Ccl5 −/− , and Ccr5 −/− mice were i.n. instilled with 8 μg of papain and 3 μg OVA-AF488 and harvested 24 h later. Flow plots illustrate myeloid cells plotted as Ly6C vs. OVA + AF488 + to gate on Ag + monocytes and DCs with scatter plots showing the total number of Ag + monocytes. Data represent two independent experiments with three to four mice per group. (F) Flow plot shows IL10 expression of LN monocytes 24 h after CpG stimulation. Histogram monocyte overlays illustrate IL10 reporter expression of monocytes stimulated with different TLR agonists. (G) Control mice received no CpG-OVA prior to sensitization with Alum + OVA and challenge with OVA. Ccr2 −/− mice, WT mice with blocked APC migration (pertussis toxin [PTx]) or depleted of monocytes (anti-Gr1 antibody) during exposure to CpG-OVA, developed significantly more airway eosinophilia compared to CpG-OVA treated WT mice. Scatter plot shows the frequency of eosinophil migration into the airways. Data combine three independent experiments with three to five mice per group. (H) Scatter plot shows the frequency of eosinophil migration into the airways of WT mice treated with CpG-OVA ± CCR5 inhibitor (Maraviroc) followed by sensitization with alum + OVA and OVA challenge. Data combine two independent experiments with four to five mice per group. (I) Scatter plot illustrates the total OVA-specific IgE in the serum of the mice from experiment H. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, mean ± SEM, a two-tailed t test (A–D), and one-way ordinary ANOVA, with post hoc Tukey’s multiple comparison test (E–I).

Article Snippet: Serum was extracted to measure circulating OVA-specific IgE (RnD systems).

Techniques: Migration, Expressing, Control, Two Tailed Test, Comparison