Journal: JCI Insight

Article Title: CXCL10 stabilizes T cell–brain endothelial cell adhesion leading to the induction of cerebral malaria

doi: 10.1172/jci.insight.98911

Figure Lengend Snippet: (A) Schematic of experiment. WT mice were coinjected with a 1:1 mix of naive Actin-TdTomato OT-I T cells and Actin-CFP OT-I Cxcr3–/– T cells. Mice were infected with ovalbumin-expressing Plasmodium berghei ANKA (PbA-OVA) 2 hours after transfer and analyzed 8 or 9 days later by (B–H) flow cytometry and (I–L) brain multiphoton intravital microscopy (MP-IVM). (B) Percentage of WT OT-I (blue symbols) and Cxcr3–/– OT-I cells (red symbols) among total CD8+ T cells in the spleen on day 8 or 9 post-infection (p.i.). Percentage of endogenous CD8+ T cells (black symbols), WT OT-I, and Cxcr3–/– OT-I cells in the spleen stained for (C) CD25, (D) CD62L, (E) CD69, (F) VLA4, or (G) LFA1. (H) Ratio of WT/Cxcr3–/– OT-I in the spleen, blood, and brain on day 8 or 9 p.i. n ≥ 6 mice/group from 3 independent experiments. On day 8 or 9 p.i., cotransfer recipients were imaged by MP-IVM. Number of (I) WT OT-I (blue symbols) and Cxcr3–/– OT-I (red symbols) per field of view (FOV) in the brain vasculature and parenchyma and (J) new T cell attachment events, (K) new detachment events, and (L) percentage of newly attached T cells that subsequently detached from the brain vasculature were enumerated using Imaris software. n = 12 mice total from 4 independent experiments. The groups were compared using (B) Wilcoxon’s, (C–I) Friedman’s, and (J and K) Krukal-Wallis tests with Dunn’s multiple comparison test. Bars and lines represent the median in all plots.

Article Snippet: The numbers of mice/group total from 3 independent experiments were as follows: DPE-GFP→WT = 13, DPE-GFP Cxcr3 –/– → Cxcr3 –/– mice = 11, DPE-GFP→ Cxcl9 –/– = 10, DPE-GFP→ Cxcl10 –/– = 14. ( B – D ) Brain multiphoton intravital microscopy (MP-IVM) of chimeric mice on day 8 or 9 after infection. ( B ) Number of T cell tracks per field of view (FOV) was analyzed using Imaris software and ( C ) arrest coefficient was calculated using MATLAB.

Techniques: Infection, Expressing, Flow Cytometry, Intravital Microscopy, Staining, Cell Attachment Assay, Software, Comparison

Journal: JCI Insight

Article Title: CXCL10 stabilizes T cell–brain endothelial cell adhesion leading to the induction of cerebral malaria

doi: 10.1172/jci.insight.98911

Figure Lengend Snippet: Lethally irradiated WT, Cxcr3–/–, Cxcl9–/–, and Cxcl10–/– mice were reconstituted with DPE-GFP or DPE-GFP Cxcr3–/– bone marrow, as indicated. Eight weeks following reconstitution, chimeric mice were infected with Plasmodium berghei ANKA (PbA). (A) Kaplan-Meier survival curve. The numbers of mice/group total from 3 independent experiments were as follows: DPE-GFP→WT = 13, DPE-GFP Cxcr3–/–→Cxcr3–/– mice = 11, DPE-GFP→Cxcl9–/– = 10, DPE-GFP→Cxcl10–/– = 14. (B–D) Brain multiphoton intravital microscopy (MP-IVM) of chimeric mice on day 8 or 9 after infection. (B) Number of T cell tracks per field of view (FOV) was analyzed using Imaris software and (C) arrest coefficient was calculated using MATLAB. Violin plots contain box plots that display the median, 25th and 75th percentiles, and whiskers that represent the 95% confidence interval (note the median of the DPE-GFP→Cxcl10–/– chimeras is 0). Numbers of new T cell attachment and detachment events were manually counted and the (D) percentage of newly attached T cells that subsequently detached were calculated. The numbers of mice/group total from 3 independent experiments were as follows: DPE-GFP→WT = 5, DPE-GFP Cxcr3–/–→Cxcr3–/– mice = 10, DPE-GFP→Cxcl9–/– = 5, DPE-GFP→Cxcl10–/– = 5. Numbers shown just below the violin plot in C represent the total number of T cells analyzed. Groups were compared using either (A) log-rank (Mantel-Cox) test or (B–D) Kruskal-Wallis with Dunn’s multiple comparison test. Bars represent the median in all plots. BM, bone marrow.

Article Snippet: The numbers of mice/group total from 3 independent experiments were as follows: DPE-GFP→WT = 13, DPE-GFP Cxcr3 –/– → Cxcr3 –/– mice = 11, DPE-GFP→ Cxcl9 –/– = 10, DPE-GFP→ Cxcl10 –/– = 14. ( B – D ) Brain multiphoton intravital microscopy (MP-IVM) of chimeric mice on day 8 or 9 after infection. ( B ) Number of T cell tracks per field of view (FOV) was analyzed using Imaris software and ( C ) arrest coefficient was calculated using MATLAB.

Techniques: Irradiation, Infection, Intravital Microscopy, Software, Cell Attachment Assay, Comparison