Fig. 2: Induction of tolerance to a hybrid insulin peptide restrains functional effector differentiation of graft-infiltrating, antigen-specific T cells. | Nature Communications

Fig. 2: Induction of tolerance to a hybrid insulin peptide restrains functional effector differentiation of graft-infiltrating, antigen-specific T cells.

From: Antigen-specific immunotherapy with a CD4+ T cell neoepitope restrains CD8+ T cell differentiation in murine pancreatic islet grafts

Fig. 2: Induction of tolerance to a hybrid insulin peptide restrains functional effector differentiation of graft-infiltrating, antigen-specific T cells.The alternative text for this image may have been generated using AI.

a Method for generating peptide-coupled PLGA nanoparticles (NPs) by covalent conjugation of the 2.5HIP (treatment) or HEL (control) peptide. b Islet transplant and NP treatment experimental design where mice are treated IV on day -7, +1, and +7 relative to the day of transplant with 2.5 mg of HEL- or HIP-conjugated NPs. c Random blood glucose readings and survival curve of transplanted mice treated with 2.5 mg of HEL NP 3x, 2.5HIP NP 3x, or 2.5HIP NP 9x (final treatment 67 d). Survival was compared between groups using Kaplan-Meier analysis with a log-rank test and Bonferroni correction for multiple comparisons. * HEL NP 3x vs HIP NP 3x, p = 0.0303; HEL NP 3x vs HIP NP 9x, p = 0.0303; HIP NP 3x vs HIP NP 9x, p = 0.0201. d Representative MHC-tetramer staining with 2.5HIP-loaded I-Ag7 or IGRP-loaded H-2kd in islet grafts. Proportion (%) and absolute number (#) of tetramer-positive 2.5HIPCD4 or IGRPCD8 T cells in islet grafts. e Representative staining of TCF1 and percentage of TCF1hi on graft-infiltrating 2.5HIPCD4 or IGRPCD8 T cells. CD44loCD62Lhi bulk, splenic CD8 T cells used as naïve controls. f Absolute number (#) of TCF1lo and TCFhi 2.5HIPCD4 or IGRPCD8 T cells in islet grafts. g, h Representative staining of inhibitory receptors TIM3 and PD1 and proportion of each IR on TCF1lo and TCF1hi, 2.5HIPCD4 and IGRPCD8 T cells. Representative staining of TCF1 by TIM3 on (i) 2.5HIPCD4 and (k) IGRPCD8 T cells. Representative cytokine staining (IFNγ, TNF) on (1) TCF1hiTIM3lo, (2) TCF1loTIM3lo, and (3) TCF1loTIM3hi T cells i 2.5HIPCD4 and (k) IGRPCD8 T cells. Proportion (%) of IFNγ+TNF+ double-positive j 2.5HIPCD4 and l IGRPCD8 T cells. In d,e,f, data are depicted as the mean ± s.e.m., with p-values obtained from a two-sided unpaired Student’s t-test. In (f–l), data are depicted as the mean ± s.e.m. with p-values obtained from one-way ANOVA with Šídák’s multiple comparison test. In (d), n = 4 mice per group for HEL NP and HIP NP. In (e), n = 3 for HEL NP 2.5HIPCD4 TCF1hi% and n = 4 for HIP NP 2.5HIPCD4 TCF1hi%, while n = 4 for HEL NP IGRPCD8 TCF1hi% and n = 5 for HIP NP IGRPCD8 TCF1hi%. In (f), n = 3 for HEL NP 2.5HIPCD4 TCF1lo/hi# and n = 4 for HIP NP 2.5HIPCD4 TCF1lo/hi#, with n = 4 for HEL NP IGRPCD8 TCF1lo/hi# and n = 5 for HIP NP IGRPCD8 TCF1lo/hi #. In (g), n = 4 mice per group for 2.5HIPCD4 TIM3% and PD1% in HEL NP and HIP NP treated mice. In (h), n = 4 mice per group for IGRPCD8 TIM3% and PD1% in HEL NP and HIP NP treated mice. In (i), n = 3 mice per group for HEL NP and HIP NP. In (i), n = 3 for HEL NP and HIP NP. In (d–f), the data are from 6 independent experiments; in (g, h, j, l), the data are from 4 independent experiments. Grey shading indicates TCF1lo T cells.

Back to article page