Figure 5.
IFNAR blocking increases DC expression of CXCR3 ligands following immune challenge. (A–C) Analysis of intrinsic IFNAR signaling on the differentiation of CD8+ T cells. Data are representative of two individual experiments with five mice in each group. Each dot in C represents a single mouse. (A) Experimental scheme. Wild-type or Ifnar−/− P14 cells were transferred into Ifnar−/− host mice prior to acute LCMV infection and analysis at d8. (B) Representative flow cytometry plots. (C) Average frequency ± SEM of TSCM cell populations. (D–G) REX3 reporter expression in cDC1, cDC2, and moDC populations during acute LCMV infection in control and d0–1 IFNAR-blocked mice. Data are representative of three individual experiments with three to four mice in each group in each experiment. Data are the mean ± SEM. (D and F) (D) d4 and (F) d8 post infection representative plots showing Cxcl9-RFP and Cxcl10-BFP reporter expression in indicated DC subsets. (E and G) Graphs summarizing graded frequencies of Cxcl9-RFP and Cxcl10-BFP expression from D and F, respectively. Graded expression summarized in key. (H) LSFM micrographs of intact REX3 lymph nodes at d8 of acute LCMV infection in control and d0–1 IFNAR-blocked conditions. Images are 200-μm longitudinal slices through the lymph node center. Scale bars represent 100 μm. The dashed line indicates the lymph node outline. Pseudocolor FIRE LUT heat maps for each REX3 reporter (left and middle panels). Merged images (right panels) show Cxcl9-RFP (magenta), Cxcl10-BFP (cyan), B220 (B cells; yellow), and CD31 (vessels; white). Images are representative of two individual experiments with four mice in each group in each experiment. (I) Detection of CXCL9 and CXCL10 protein staining in cDC1, cDC2, and moDC populations at d4 of acute LCMV infection. Graphs show average gMFI ± SEM. The dashed line indicates chemokine protein detection in indicated DC subsets from Cxcl9−/− and Cxcl10−/− mice. Data are representative of three individual experiments with at least three mice in each group in each experiment. Each dot represents a single mouse. Statistics was determined using unpaired t tests. *P < 0.05, **P < 0.01, ***P < 0.001. Fig. S3 shows additional supporting data.

IFNAR blocking increases DC expression of CXCR3 ligands following immune challenge. (A–C) Analysis of intrinsic IFNAR signaling on the differentiation of CD8+ T cells. Data are representative of two individual experiments with five mice in each group. Each dot in C represents a single mouse. (A) Experimental scheme. Wild-type or Ifnar−/− P14 cells were transferred into Ifnar−/− host mice prior to acute LCMV infection and analysis at d8. (B) Representative flow cytometry plots. (C) Average frequency ± SEM of TSCM cell populations. (D–G) REX3 reporter expression in cDC1, cDC2, and moDC populations during acute LCMV infection in control and d0–1 IFNAR-blocked mice. Data are representative of three individual experiments with three to four mice in each group in each experiment. Data are the mean ± SEM. (D and F) (D) d4 and (F) d8 post infection representative plots showing Cxcl9-RFP and Cxcl10-BFP reporter expression in indicated DC subsets. (E and G) Graphs summarizing graded frequencies of Cxcl9-RFP and Cxcl10-BFP expression from D and F, respectively. Graded expression summarized in key. (H) LSFM micrographs of intact REX3 lymph nodes at d8 of acute LCMV infection in control and d0–1 IFNAR-blocked conditions. Images are 200-μm longitudinal slices through the lymph node center. Scale bars represent 100 μm. The dashed line indicates the lymph node outline. Pseudocolor FIRE LUT heat maps for each REX3 reporter (left and middle panels). Merged images (right panels) show Cxcl9-RFP (magenta), Cxcl10-BFP (cyan), B220 (B cells; yellow), and CD31 (vessels; white). Images are representative of two individual experiments with four mice in each group in each experiment. (I) Detection of CXCL9 and CXCL10 protein staining in cDC1, cDC2, and moDC populations at d4 of acute LCMV infection. Graphs show average gMFI ± SEM. The dashed line indicates chemokine protein detection in indicated DC subsets from Cxcl9−/− and Cxcl10−/− mice. Data are representative of three individual experiments with at least three mice in each group in each experiment. Each dot represents a single mouse. Statistics was determined using unpaired t tests. *P < 0.05, **P < 0.01, ***P < 0.001. Fig. S3 shows additional supporting data.

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