Fig. 6. pKL cells revert hyperglycemia in NOD mice in vivo.

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Fig. 6. pKL cells revert hyperglycemia in NOD mice in vivo. pKL cells revert hyperglycemia in NOD mice in vivo. (A and B) Newly hyperglycemic NOD mice were treated with WT cells, pKL cells, or Tg cells. Data are representative of 10 untreated mice, 10 mice treated with WT cells, 10 mice treated with pKL cells, and 15 mice treated with Tg cells. Incidence of diabetes in all groups of NOD mice (untreated, WT-treated, pKL-treated, and Tg-treated mice) was compared using the log-rank (Mantel-Cox) test; #P < 0.0001 versus all, P < 0.0001 for Tg versus WT, and P < 0.05 for pKL versus WT. (C) Quantification of IFN-γ–producing cells in an in vitro assay, in which splenocytes isolated from newly hyperglycemic pKL-treated NOD mice or in untreated hyperglycemic NOD mice 40 days after treatment were challenged with islet peptides after treatment with pKL, normalized for background. Experiments were run in triplicate, and statistical analysis was performed using two-tailed unpaired t test. (D) Immunophenotype of lymphocytes isolated from spleen of pKL-treated newly hyperglycemic NOD mice showed an increase in the percentage of FoxP3+ regulatory T cells. Experiments were run in triplicate (n = 3 pKL-treated and n = 5 untreated), and statistical analysis was performed using two-tailed unpaired t test. (E to G) Bar graph showing the levels of pro- (IL-2/IL-6) and anti-inflammatory (IL-4) cytokines as measured by Luminex in the serum of untreated or pKL-treated newly hyperglycemic NOD mice at baseline and at 7 days after treatment. Experiments were run in triplicate per group, and condition (D0 and D7) and statistical analysis were performed using two-tailed unpaired t test. (H) OVA rechallenge. Experiments were run in triplicate per group, and statistical analysis were performed using two-tailed unpaired t test, #P < 0.05 versus all. (I) Insulitis score in untreated and pKL-treated newly hyperglycemic NOD mice. (J and K) Representative immunohistochemical H&E analysis and CD3/insulin staining in serial pancreatic islet tissue sections from pKL-treated or untreated NOD mice; n = 9 sections per group were analyzed. Histology magnification, ×20. Scale bars, 200 μm. Data are expressed as means ± SEM. Data are representative of at least n = 3 mice. *P < 0.05; **P < 0.01; ***P < 0.0001; #P < 0.05 versus all. Moufida Ben Nasr et al., Sci Transl Med 2017;9:eaam7543 Published by AAAS