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. 2017 Oct 16;8:951. doi: 10.1038/s41467-017-00982-x

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — PI3Kγ or PI3Kδ deletion and long-term allograft survival. a single dose CTLA4-Ig, injected into PI3Kγ ^−/− recipients induced long-term acceptance of fully mismatched BALB/c cardiac allografts compared to the controls (MST of 41 vs. >100 days, *p < 0.05, t-test, n = 10/group). However, single dose CTLA4-Ig, injected into PI3Kδ ^D910A/D910A recipients of fully mismatched BALB/c cardiac allografts showed reduced allograft survival compared to the controls (MST: 41 vs. 32 days, *p < 0.05, t-test, n = 7–10/group). b Representative examples of cardiac allograft histology at day 100 post-transplant show lower grades of acute cellular rejection in the PI3Kγ ^−/− hearts treated with single dose CTLA4-Ig compared to cardiac allograft at day 28 post-transplant from WT mice treated with single dose CTLA4-Ig (n = 4/group). Inset show worse vasculopathy in the WT recipients treated with single dose CTLA4-Ig compared to PI3Kγ ^−/− hearts treated with single dose CTLA4-Ig (H&E stain, scale bar 50 μm, inset scale bar 25 μm). Immunohistochemistry analysis of CD3 and FoxP3 shows fewer CD3+ infiltrating cells in the PI3Kγ ^−/− hearts treated with single dose CTLA4-Ig compared to WT but relatively more FoxP3 (scale bar 50 μm). c Representative dot plots of leukocytes isolated from the allograft of PI3Kγ ^−/− and WT recipients treated with single dose CTLA4-Ig show significantly less CD4⁺ and CD8⁺ infiltration in the PI3Kγ ^−/− allografts compared to WT. Bar graph represents the absolute count of CD4⁺ and CD8⁺ T cells in the heart allografts of PI3Kγ ^−/− and WT recipients treated with single dose CTLA4-Ig. (*p < 0.05, t-test, n = 6/group, the graphs show data as mean ± s.e.m.). d Representative examples of cardiac allograft histology at day 28 post-transplant show higher grades of acute cellular rejection in the PI3Kδ ^D910A/D910A hearts treated with single dose CTLA4-Ig compared to WT mice treated with single dose CTLA4-Ig (n = 4/group). Inset show more severe vasculitis in the PI3Kδ ^D910A/D910A treated with single dose CTLA4-Ig compared to WT-recipient hearts treated with single dose CTLA4-Ig (H&E stain, scale bar 50 μm, inset scale bar 25 μm). (MST mean survival time)