Figure 3.

iPS-DCreg Immunization Induces Permanent Acceptance of Allogeneic Cardiac Grafts and Decreases CD8⁺ T Cells in Grafts and Spleen

(A) 1 × 10⁶ B6 DCs were injected intravenously into CBA mice 7 days before heterotopic cardiac transplantation. A statistical evaluation of graft survival was performed using Kaplan-Meier curves and compared using log-rank tests. ^∗p < 0.05, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001.

(B) Cardiac allografts of POD7 and POD14 were stained with H&E. The red arrow indicates the increased lymphocytic infiltration around the coronary artery in PBS treated allograft.

(C) Allografts were stained with anti-CD8 (blue), collagen IV (yellowish-brown), and BrdU (red) by triple immunostaining. The black arrows indicate the CD8 and BrdU double positive lymphocytes in the grafts.

(D) Spleen cells (SPCs) were harvested on POD7 and POD14 for CD8⁺ T cell assessment by FCM analysis (n = 4 in each group, mean ± SD, pooled from four independent experiments). ^∗p < 0.05.

(E) The mRNA expression of GranzymeB and Perforin, IFN-γ, TNF-α, IL-1β, and IL-6 in allografts was measured by qRT-PCR (n = 9 in the PBS-POD7 group, n = 6 in the iPS-DCregs POD7 group, n = 5 in the iPS-DCregs POD14 group; mean ± SD, pooled from five independent experiments). Statistical analysis was determined by one-way ANOVA and Tukey's test. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.