Fig. 6: LiNKTR1 cytokine-mediated suppression of local and proximal APCs and diabetogenic autoimmunity without blunting normal immune responses.
a, b Macroscopic (a) and microscopic (b) PBC scores of control NOD.c3c4 mice and NOD.c3c4 mice treated with αGalCer/CD1d-NPs along with rat-IgG or blocking mAbs against mouse IL-4, IL-10, IL-21Rα or IFNγ. In a, data correspond to n = 19 control mice (1 untreated and 18 Cys-NP-treated mice, pooled; 6 experiments) and n = 20 (6 experiments), 4, 6 (2 experiments), 4 and 3 mice treated with NPs and mAbs (from left to right). In b, data correspond to n = 21 control mice (1 untreated and 18 Cys-NP-treated mice, pooled; 6 experiments) and n = 23 (6 experiments), 4, 8 (3 experiments), 5 and 3 mice treated with NPs and mAbs (from left to right). All data were compared to the pMHCII-NP-treated group. c BrdU incorporation by splenic 8.3-CD8 + T cells in response to NRP-V7 peptide-pulsed liver KCs from NOD.c3c4 mice, in the absence or presence of rat-IgG, or blocking mAbs against mouse IL-4, IL-10, IL-21Rα or IFNγ, and LiNTKs from control (Cys-NP-treated) or αGalCer/CD1d-NP-treated NOD.c3c4 mice. Data correspond to n = 4, 4, 5, 5, 5, and 5/group. d IFNγ secretion by 8.3-CD8 + T cells from 8.3-NOD.G6pc2–/–.Tcra–/– mice upon stimulation with NRP-V7- or TUM-peptide-pulsed CD11b + APCs isolated from the PCLNs of control NOD mice (treated with Cys-NPs (n = 3) or αGalCer/CD1d-NP-treated NOD mice (n = 3). e, f In vivo proliferation of naïve CFSE + 8.3-CD8 + T cells from 8.3-NOD.G6pc2–/–.Tcra–/– donors in the PCLN, PLN, MLN and spleen of control NOD (n = 5–6/organ; top in f) and control (n = 5, treated with Cys-NPs) or αGalCer/CD1d-NP-treated NOD.c3c4 mice (n = 6; bottom in f). e shows representative plots. g Evolution of blood glucose levels of acutely diabetic mice (>11 mM) in response to vehicle (n = 5) vs. αGalCer/CD1d-NP treatment (n = 4). Horizontal arrows show the duration of treatment with αGalCer/CD1d-NPs. h EAE scores (top) and body weight changes (bottom) of B6 mice with EAE in response to Cys-NP- (n = 9), αGalCer/CD1d-NP- (n = 10) or MOG38-49/IAb-NP (n = 6) treatment. Data correspond to two independent experiments. i Average percentages of LiNKTR1 cells (cluster #5) in liver, PLNs and pancreatic islets as determined via mass cytometry. Data correspond to 5 mice/organ. j Left: average percentages of InsB13-21/I-Ag7 tetramer+ cells in islet-associated CD4 + T cells of NOD mice treated with αGalCer/CD1d-NP (n = 4) or control NPs (n = 3); Right: representative FACS profile. k Bacterial load of L. monocytogenes in the spleen and liver 3, and 35 days after infection (n = 4, 4, and 5; left to right). l Serum anti-DNP antibody titers upon KLH–DNP immunization (n = 3, 5 and 5; left to right). Data correspond to means ± SEM. P values were calculated using one-way ANOVA with Dunnett’s post hoc correction (a–c, f, l), one tail Mann–Whitney U (d, i, j, k), or two-way ANOVA (g, h). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.
