Anti-CD3 mAb treatment reshapes infiltrating T and β cells in the islets in autoimmune diabetes
Ying Wu, Maxwell Spurrell, Ana Lledó-Delgado, Songyan Deng, Dejiang Wang, Yang Liu, Mahsa Nouri Barkestani, Ana Luisa Perdigoto, Kevan C. Herold
Ying Wu, Maxwell Spurrell, Ana Lledó-Delgado, Songyan Deng, Dejiang Wang, Yang Liu, Mahsa Nouri Barkestani, Ana Luisa Perdigoto, Kevan C. Herold
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Research Article Endocrinology Immunology

Anti-CD3 mAb treatment reshapes infiltrating T and β cells in the islets in autoimmune diabetes

Abstract

Treatment with anti-CD3 monoclonal antibody (mAb) can delay or prevent type 1 diabetes in mice and humans by modulating the immune-mediated destruction of β cells. A single course of treatment may have lasting efficacy, but the mechanisms that account for these prolonged effects, i.e., “operational tolerance,” are not clear. Here, we used paired single-cell RNA and T cell receptor sequencing to characterize islet-infiltrating T cells and their counterpart in paired pancreatic lymph nodes from anti-CD3 mAb–treated nonobese diabetic (NOD) mice in remission. We found that after anti-CD3 mAb treatment, T cells that infiltrate the islets are more heterogeneous and have hybrid features including characteristics of T stem cell–like memory and reduced effector function compared with those from untreated prediabetic NOD mice. Autoantigen-reactive CD8+ T cells persist after treatment, but they also show features of stemness and reduced pathogenicity. Our findings describe the reshaping of islet-infiltrating and autoreactive T cells and β cells that lead to operational, but tenuous, tolerance to autoimmune diabetes following anti-CD3 mAb treatment.

Authors

Ying Wu, Maxwell Spurrell, Ana Lledó-Delgado, Songyan Deng, Dejiang Wang, Yang Liu, Mahsa Nouri Barkestani, Ana Luisa Perdigoto, Kevan C. Herold

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Figure 4

Changes in intra-islet CD8+ T cells after anti-CD3 mAb treatment.

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Changes in intra-islet CD8+ T cells after anti-CD3 mAb treatment. (A) UM...
(A) UMAP showing CD8+ T cells in islets or ppLNs of anti-CD3–treated remitter and prediabetic NOD mice, colored by conditions. (B) Volcano plot showing DEGs of islet-infiltrating CD8+ T cells between remitter and prediabetic NOD (adjusted P values < 0.05). Upregulated genes of note in remitters are highlighted in red and downregulated genes in blue. (C) Bar chart showing percentage of granzyme B+ islet-infiltrating CD8+ T cells of anti-CD3–treated versus untreated prediabetic NOD mice by flow cytometry. Each symbol represents an individual mouse (**P = 0.008, unpaired, 2-tailed t test). (D) Ingenuity Pathway Analysis of islet-infiltrating CD8+ T cells in remitter versus prediabetic NOD mice (z scores ≤ –2, P values < 0.05). (E) Bar graphs showing TCR clonotype frequencies of CD8+ T cells. Raw count of each clonotype was divided by total count, and result is shown with normalization by scaling to 1. Each bar represents 1 individual mouse. Colors denote clonotype size based on raw count. (F) Repertoire diversity of CD8+ T cells was measured by Shannon entropy index (entropy index scaled to 1; data shown as mean ± SEM). Each symbol represents 1 individual mouse (*P = 0.038, ****P < 0.0001, 2-way ANOVA with Tukey’s multiple-comparison test). (G and H) Spatial-ATAC-seq of islets from remitter and prediabetic mice. (G) H&E images feature 1 representative islet of either condition with similar degree of insulitis (original magnification, ×10). (H) Summary bar charts showing gene scores for Tcf7 expression (a total of 10 islets from 3 remitter NOD mice and a total of 8 islets from 3 prediabetic NOD mice evaluated by 3 repeated measurements with mixtures of islets from both groups; *P = 0.034, Mann-Whitney test). Data are shown as mean ± SEM.