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of Florida"],"full_dataset_link":["https://www.ebi.ac.uk/ena/browser/view/PRJNA1070313"],"scientific_name":["Mus musculus"],"long_description":["Immunotherapeutics that modulate T cell activation represent a crucial component for inhibiting the autoimmune pathogenesis of type 1 diabetes (T1D) and improving the efficacy of β-cell replacement therapy. Here, we present a novel strategy for reducing diabetes incidence in the NOD mouse using monoclonal antibodies (mAbs) to block the T cell costimulatory receptor, CD226. Notably, female NOD mice treated with 600 µg of ⍺-CD226 mAbs between 7-8 weeks of age showed decreased disease incidence at 30 weeks and reduced insulitis severity compared to mice treated with an isotype control. Ex vivo analysis performed five weeks post-treatment revealed ⍺-CD226 mAbs persist in vivo, reducing the availability of CD226 on CD8+ T cells and Tregs. Flow cytometric analysis demonstrates that ⍺-CD226 mAbs inhibit the proliferation of both CD4+ and CD8+ T cells in vitro. Similarly, ex vivo samples had reduced CD4+ and CD8+ effector memory T cell proliferation following ⍺-CD226 mAb treatment. Splenocytes treated with ⍺-CD226 mAbs exhibited a more immunoregulatory cytokine profile with decreased IFN-γ and increased IL-10 production. This phenotype was further corroborated by 51Cr-release assays demonstrating reduced cell-mediated lympholysis (CML) of murine β-cells by ⍺-CD226 mAb-treated autoreactive cytotoxic lymphocytes. Ex vivo phenotyping of FOXP3+Helios+ Tregs revealed increased CD25 expression following ⍺-CD226 mAb treatment, with Tregs displaying augmented suppressive capacity of CD4+ responders during in vitro suppression assays. These data suggest that ⍺-CD226 mAbs both reduce T cell cytotoxicity and improve Treg function, with important therapeutic implications for the prevention or suspension of T1D. Overall design: NOD mice received three doses of either ⍺-CD226 mAb or Isotype Control between 7-8 weeks of age. At 12 weeks of age, CD3+ T cells enriched from pancreatic-draining lymph nodes (pLN) and collagenase-digested pancreas were stained with ADT antibodies before generating gene expression, V(D)J and surface protein libraries. ADT: Barcode Sequence TotalSeq-C0001 anti-mouse CD4: AACAAGACCCTTGAG TotalSeq-C0002 anti-mouse CD8⍺: TACCCGTAATAGCGT TotalSeq-C0073 anti-mouse CD44: TGGCTTCAGGTCCTA TotalSeq-C0112 anti-mouse CD62L: TGGGCCTAAGTCATC TotalSeq-C0848 anti-mouse TIGIT: GAAAGTCGCCAACAG"],"tag":["xref:PubMed:39636437","xref:PubMed:39071293"],"repository":["ENA"],"additional_accession":[]},"is_claimable":false,"name":"Monoclonal Antibody Blockade of CD226 Decreases Spontaneous Diabetes in the NDO Mouse by Augmenting Treg and Diminishing Effector Functionality","description":"Monoclonal Antibody Blockade of CD226 Decreases Spontaneous Diabetes in the NDO Mouse by Augmenting Treg and Diminishing Effector Functionality","dates":{"last_updated":"2025-09-24","first_public":"2024-08-26"},"accession":"PRJNA1070313","cross_references":{"GEO":["GSE254403"],"taxon":["10090"],"PubMed":["39071293","39636437"]}}