Protection of Islet Grafts Through Transforming Growth Factor-β–Induced Tolerogenic Dendritic Cells

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FIG. 6.
FIG. 6.

Decreased activation of DCs and lower proliferation of islet antigen–specific T cells in draining lymph nodes of TGF-β–expressing grafts. Graft-draining RLNs (draining RLNs [dRLNs]) and control RLNs (cRLNs) were isolated from mice that had received a TGF-β–expressing or WT islet graft 5 days previously. DCs were CD11c+CD37AAD and assessed for expression of CD80 (top), CD86 (middle), and MHC II (bottom) (A). To the left are representative flow cytometry histogram overlays comparing WT grafts (black histograms) with TGF-β–expressing grafts (white histograms) in cRLNs and dRLNs. To the right are graphs showing the relative decrease in expression in the repeat experiments (separate biological replicates; each dot represents the data obtained from one mouse), with the value from the WT graft set as 100% (12 mice for CD80, 10 mice for CD86, and 8 mice for MHC II). Differences between groups were assessed using the Mann-Whitney test (A). Filled circles, control grafts; open circles, TGF-β–producing grafts. CFSE-labeled insB15-23–specific CD8+ T cells were transferred into CD45 congenic RIP-TNF NOD mice having received either a TGF-β–expressing graft or a WT graft 2 days previously. Seventy-two hours later, the draining and nondraining lymph nodes were harvested and proliferation was assessed as dilution of CFSE signal in the CD45.1+CD45.2CD8+7AAD gate (B). The experiment was repeated four times, with a compilation of all four experiments (eight mice) at the bottom and representative fluorescence-activated cell sorter plots depicted above. Differences in proliferation between the draining RLNs of WT and TGF-β–expressing grafts were determined using a paired Student t test. *P ≤ 0.05, **P ≤ 0.01.

This Article

  1. Diabetes vol. 62 no. 9 3132-3142