Fig 2: Dendritic cell (DC) populations in omental tumors. (a) Representative gating for MHC-II+CD11chi DC populations in tumor-naive or tumor-bearing mice with treatment with vehicle control or WNT974 for 14 days. (b) Representative gating for CD103+ DCs treated with vehicle control (top) or WNT974 (bottom). (c) Graphical representation data from plots in panels (b) and (c). Percentage frequency of parent shown on the left, total number of cells shown on the right (n = 5–7 mice/group, data from one of three independent experiments, dots represent individual mice).

Fig 3: Integrin αE deficiency leads to a fecal IgA deficit despite normal B cell recruitment, IgA production and increased pIgR mRNA expression.a Fecal IgA levels measured by ELISA in C57BL/6 (WT), Itgb7−/− (β7−/−), Itgae−/− (αE−/−) and pIgR-deficient (pIgR−/−) mice. b Percentage of CD3+ and CD19+ cells within the ileal LP of indicated strains and representative contour plots of indicated cell subsets gated on live, single cellular events. c IF staining of IgA + ASC and E-cadherin, representative coronal images of terminal ileum (TI). d Absolute numbers and representative dot plots of IgA + ASCs in lamina propria. e LP IgA levels measured by ELISA of indicated strains (f) ileal IgA mRNA transcripts in ilea of indicated mice (g) pIgR mRNA expression in ileum of indicated mice. h Relative pIgR fluorescence intensity expressed as integrated density (IntDent, see “Methods”) per villi and (i) representative IF images (j) Percentage of IgA-coated fecal bacteria measured by flow cytometry and (j, k, l) representative plots and histograms of IgA coating of indicated mouse strains. Each data point (mean ± SD) represent a single mouse with n ≥ 5 from 2 or 3 independent experiments. Statistical significance determined using two-way ANOVA with Dunnett’s multiple comparisons test.

Fig 4: GFPhi Cells during Influenza Virus Infection Are Infected Ly6Chi Monocytes(A and B) Mx1gfp mice were infected with 102 pfu of PR8, and GFP expression was measured in T cells (CD3+), B cells (B220+), NK cells (NK1.1 +), alveolar macrophages (CD11c+SiglecF+), CD11b DCs (CD11chiMHCIIhiCD11b+CD103−), CD103 DCs (CD11chiMHCIIhiCD103+CD11b−), neutrophils (Ly6C+Ly6G+CD11b+), and Ly6Chi monocytes (Ly6ChiLy6G−CD11b+) at day 7 post-infection.(C and D) Mx1gfp mice were infected with 104 pfu of PR8 and stained for surface M2 protein in T cells, B cells, Ly6Chi monocytes, or epithelial cells at day 3.(B) shows the mean ± SD for n = 4–6 animals. (D) shows the mean ± SD for 3 animals. (A) and (C) show representative FACs plots. See also Figure S5.

Fig 5: AHR signaling promotes TRM differentiation and function in human CD8 T cells(A–C) Human peripheral blood (PBMC) vs. IEL CD8+ T cells were analyzed via flow cytometry. Staining of CD45RA and CD45RO (top), CD103 and CD69 (bottom) (A), as well as T-BET and AHR (B) in tissue fractions as indicated in the figure. Quantification of AHR protein levels (gMFI) in human PBMC, LPL, and IEL CD8+ T cells (C). Data are compiled from three independent experiments and shown as mean ± SEM (n = 4 replicates per group).(D–G) Human naive CD8+ T cells were isolated from PBMCs and then subjected to in vitro TRM-like differentiation culture conditions. The cells were given differentiation cytokines and treated with DMSO (control), FICZ, or CH223191 on day 2. The assay was collected on day 4, and flow cytometry quantification of CD103 expression was performed (D). Data are compiled from three independent experiments and are shown as mean ± SEM (n = 4 replicates per group). RNA was isolated for qRT-PCR expression analysis of AHR (E), AHR direct target gene AHRR (F), and GZMB (G). Data are shown as mean ± SEM (n = 3 technical replicates per group). Data are representative of three independent experiments.