Fig. 4
PI3Kδ deletion abrogates the tolerogenic effect of CTLA4-Ig. a Multiple dose CTLA4-Ig treatment of PI3Kδ D910A/D910A recipients abrogates the long-term acceptance of fully mismatched BALB/c cardiac allografts seen in treated WT controls (MST of > 100 vs. 96 days, *p < 0.05, t-test, n = 10/group). b Representative examples of cardiac allograft histology harvested 100 days after transplant show higher grades of cellular infiltration (H&E stain, scale bar 50 μm) and vasculopathy (H&E stain, scale bar 25 μm) in the PI3Kδ D910A/D910A hearts treated with multiple dose CTLA4-Ig compared to WT mice treated with multiple dose CTLA4-Ig (n = 4/group). The bar graphs show the average of lymphocyte infiltration score and vasculopathy score. The PI3Kδ D910A/D910A hearts treated with multiple dose CTLA4-Ig shows significantly higher score compared to WT mice treated with multiple dose CTLA4-Ig (*p < 0.05, t-test, n = 4/group, the graphs show data as mean ± s.e.m.). c Representative examples of dot plots of leukocytes isolated from the allograft of PI3Kδ D910A/D910A and WT recipients treated with multiple dose CTLA4-Ig shows more CD4+ and CD8+ infiltration in the allografts of PI3Kδ D910A/D910A recipients compared to WT control. Graph represents the absolute count of graft-infiltrating CD4 and CD8 in PI3Kδ D910A/D910A recipients treated with multiple dose CTLA4-Ig compared to WT control treated with multiple dose CTLA4-Ig (*p < 0.05, t-test, n = 6/group). d Representative examples of dot plots of Tregs in the DLN of PI3Kδ D910A/D910A and WT recipients treated with multiple dose CTLA4-Ig show significantly fewer Tregs in the PI3Kδ D910A/D910A recipients compared to WT control (*p < 0.05, t-test, n = 6/group). e Representative examples of histograms of GrB expression by CD8+ CD44high T cells in the spleen of PI3Kδ D910A/D910A and WT recipients treated with multiple dose CTLA4-Ig show significantly more GrB expression in CD8+ of the PI3Kδ D910A/D910A recipients compared to WT (*p < 0.05, t-test, n = 4/group). f WT and PI3Kδ D910A/D910A recipient splenocytes were stimulated with donor splenocytes. Bar graphs represent the level of IFNγ, IL-6, IL-17, and IP10 in the supernatant collected from the MLR assay as measured by luminex (*p < 0.05, t-test, n = 4/group). c–f The graphs show data as mean ± s.e.m. (MST mean survival time; DLN draining lymph node; Treg regulatory T cell; GrB GranzymeB)
