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A TLR7 ligand prevents mouse GvHD
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Figure 4. R848 treatment lowers effector T cells but increases regulatory T cells during non-conditioned graft-versus-host disease. (A) B6D2F1 and B6 mice, treated with R848 or not, received 15x106 B6 CD5+ cells labeled with CFSE from B6 mice not treated (NT) or treated with R848 48 h before cell transfer. After 6 days, anti- H-2Dd, -H-2Db and -TCRβ antibodies were used to analyze the proliferation level of B6 T cells in host spleen by flow cytometry. (B-E) 14 days after total B6 spleen cell transfer, spleen cell subsets were enumerated by FACS, using anti-H2Dd, -H2Db, -CD4, -CD8, -CD44, -CD62L and -CD69 antibodies. (B) Plots show absolute numbers of CD4 and CD8 T cells. (C) CD44, CD62L and (D) CD69 expression by T cells is represented with density plots or histograms for percentages and plots for absolute numbers. (E) H-2Dd, H-2Db, LIVE/DEAD®, CD4, TCRβ and Foxp3 staining was used to determine by flow cytometry the percentage (left) and absolute number (right) of Treg in host spleen 14 days after induction of GvHD. Data are from two to three experiments in all panels (*P<0.05, **P<0.01, ***P<0.001 by the Kruskal-Wallis test with Dunn multiple comparison test or Mann–Whitney unpaired t-test).
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