{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T04:23:02Z","timestamp":1790310182823,"version":"4.1.0"},"reference-count":63,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,4,4]],"date-time":"2022-04-04T00:00:00Z","timestamp":1649030400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100008871","name":"Juvenile Diabetes Research Foundation United States of America","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100008871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>Antigen-specific therapies that suppress autoreactive T cells without inducing systemic immunosuppression are a much-needed treatment for autoimmune diseases, yet effective strategies remain elusive. We describe a microfluidic Cell Squeeze<jats:sup>\u00ae<\/jats:sup> technology to engineer red blood cells (RBCs) encapsulating antigens to generate tolerizing antigen carriers (TACs). TACs exploit the natural route of RBC clearance enabling tolerogenic presentation of antigens. TAC treatment led to antigen-specific T cell tolerance towards exogenous and autoantigens in immunization and adoptive transfer mouse models of type 1 diabetes (T1D), respectively. Notably, in several accelerated models of T1D, TACs prevented hyperglycemia by blunting effector functions of pathogenic T cells, particularly in the pancreas. Mechanistically, TACs led to impaired trafficking of diabetogenic T cells to the pancreas, induced deletion of autoreactive CD8 T cells and expanded antigen specific Tregs that exerted bystander suppression. Our results highlight TACs as a novel approach for reinstating immune tolerance in CD4 and CD8 mediated autoimmune diseases.<\/jats:p>","DOI":"10.3389\/fimmu.2022.869669","type":"journal-article","created":{"date-parts":[[2022,4,4]],"date-time":"2022-04-04T12:40:53Z","timestamp":1649076053000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":20,"title":["Engineered RBCs Encapsulating Antigen Induce Multi-Modal Antigen-Specific Tolerance and Protect Against Type 1 Diabetes"],"prefix":"10.3389","volume":"13","author":[{"given":"Colin J.","family":"Raposo","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Judith D.","family":"Cserny","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gloria","family":"Serena","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan N.","family":"Chow","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia","family":"Cho","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanyang","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Kotler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Armon","family":"Sharei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Howard","family":"Bernstein","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shinu","family":"John","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,4,4]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1016\/bs.ircmb.2018.05.008","article-title":"Chapter Three T Lymphocytes and Autoimmunity","volume":"341","author":"Khan","year":"2018","journal-title":"Int Rev Cel Mol Bio"},{"key":"B2","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1016\/j.coi.2021.03.019","article-title":"Manipulating Antigen Presentation for Antigen-Specific Immunotherapy of Autoimmune Diseases","volume":"70","author":"Streeter","year":"2021","journal-title":"Curr Opin Immunol"},{"key":"B3","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1111\/j.1600-065x.2011.01019.x","article-title":"Mechanisms of Tolerance","volume":"241","author":"Bluestone","year":"2011","journal-title":"Immunol Rev"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1056\/nejmra1911109","article-title":"Tolerance in the Age of Immunotherapy","volume":"383","author":"Bluestone","year":"2020","journal-title":"New Engl J Med"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-019-0015-4","article-title":"Antigen-Specific Therapeutic Approaches for Autoimmunity","volume":"37","author":"Serra","year":"2019","journal-title":"Nat Biotechnol"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1038\/nri2153","article-title":"Antigen-Specific Tolerance Strategies for the Prevention and Treatment of Autoimmune Disease","volume":"7","author":"Miller","year":"2007","journal-title":"Nat Rev Immunol"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1126\/science.aay3638","article-title":"A Noninflammatory mRNA Vaccine for Treatment of Experimental Autoimmune Encephalomyelitis","volume":"371","author":"Krienke","year":"2021","journal-title":"Science"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1038\/s41551-019-0424-1","article-title":"Synthetically Glycosylated Antigens Induce Antigen-Specific Tolerance and Prevent the Onset of Diabetes","volume":"3","author":"Wilson","year":"2019","journal-title":"Nat BioMed Eng"},{"key":"B9","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1016\/j.it.2020.11.001","article-title":"Tolerogenic Dendritic Cells in Autoimmunity and Inflammatory Diseases","volume":"42","author":"Morante-Palacios","year":"2020","journal-title":"Trends Immunol"},{"key":"B10","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1136\/annrheumdis-2015-208456","article-title":"Autologous Tolerogenic Dendritic Cells for Rheumatoid and Inflammatory Arthritis","volume":"76","author":"Bell","year":"2017","journal-title":"Ann Rheum Dis"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.661875","article-title":"Antigen-Specific Regulatory T Cell Therapy in Autoimmune Diseases and Transplantation","volume":"12","author":"Selck","year":"2021","journal-title":"Front Immunol"},{"key":"B12","doi-asserted-by":"publisher","first-page":"398","DOI":"10.1038\/s41580-020-0232-1","article-title":"The Clearance of Dead Cells by Efferocytosis","volume":"21","author":"Boada-Romero","year":"2020","journal-title":"Nat Rev Mol Cell Bio"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1098\/rsos.160850","article-title":"How do Red Blood Cells Know When to Die","volume":"4","author":"Arias","year":"2017","journal-title":"Roy Soc Open Sci"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2011.00057","article-title":"Modulation of Macrophage Efferocytosis in Inflammation","volume":"2","author":"Korns","year":"2011","journal-title":"Front Immunol"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2011.08.011","article-title":"Cell Death in the Maintenance and Abrogation of Tolerance: The Five Ws of Dying Cells","volume":"35","author":"Griffith","year":"2011","journal-title":"Immunity"},{"key":"B16","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1016\/j.ijpharm.2012.12.044","article-title":"Red Blood Cells as Innovative Antigen Carrier to Induce Specific Immune Tolerance","volume":"443","author":"Cremel","year":"2013","journal-title":"Int J Pharmaceut"},{"key":"B17","doi-asserted-by":"publisher","first-page":"188ra75","DOI":"10.1126\/scitranslmed.3006168","article-title":"Antigen-Specific Tolerance by Autologous Myelin Peptide\u2013Coupled Cells: A Phase 1 Trial in Multiple Sclerosis","volume":"5","author":"Lutterotti","year":"2013","journal-title":"Sci Transl Med"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1701746114","article-title":"Engineered Erythrocytes Covalently Linked to Antigenic Peptides can Protect Against Autoimmune Disease","volume":"114","author":"Pishesha","year":"2017","journal-title":"Proc Natl Acad Sci"},{"key":"B19","doi-asserted-by":"publisher","first-page":"e1500112","DOI":"10.1126\/sciadv.1500112","article-title":"Engineered Binding to Erythrocytes Induces Immunological Tolerance to E. Coli Asparaginase","volume":"1","author":"Lorentz","year":"2015","journal-title":"Sci Adv"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1216353110","article-title":"Engineering Antigens for in Situ Erythrocyte Binding Induces T-Cell Deletion","volume":"110","author":"Kontos","year":"2013","journal-title":"Proc Natl Acad Sci"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1016\/j.blre.2017.06.001","article-title":"Eryptosis in Health and Disease: A Paradigm Shift Towards Understanding the (Patho)Physiological Implications of Programmed Cell Death of Erythrocytes","volume":"31","author":"Qadri","year":"2017","journal-title":"Blood Rev"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1038\/srep15907","article-title":"Memory of Tolerance and Induction of Regulatory T Cells by Erythrocyte-Targeted Antigens","volume":"5","author":"Grimm","year":"2015","journal-title":"Sci Rep-uk"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1126\/sciimmunol.abe1801","article-title":"Persistent Antigen Exposure via the Eryptotic Pathway Drives Terminal T Cell Dysfunction","volume":"6","author":"Watkins","year":"2021","journal-title":"Sci Immunol"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1004175","article-title":"Tolerance Induced by Apoptotic Antigen-Coupled Leukocytes Is Induced by PD-L1+ and IL-10\u2013Producing Splenic Macrophages and Maintained by T Regulatory Cells","volume":"187","author":"Getts","year":"2011","journal-title":"J Immunol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1900\/rds.2012.9.319","article-title":"Tolerance Strategies Employing Antigen-Coupled Apoptotic Cells and Carboxylated PLG Nanoparticles for the Treatment of Type 1 Diabetes","volume":"9","author":"Prasad","year":"2012","journal-title":"Rev Diabetes Stud"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.178.4.2212","article-title":"Peripheral Tolerance Induction Using Ethylenecarbodiimide-Fixed APCs Uses Both Direct and Indirect Mechanisms of Antigen Presentation for Prevention of Experimental Autoimmune Encephalomyelitis","volume":"178","author":"Turley","year":"2007","journal-title":"J Immunol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1218705110","article-title":"A Vector-Free Microfluidic Platform for Intracellular Delivery","volume":"110","author":"Sharei","year":"2013","journal-title":"Proc Natl Acad Sci"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1039\/c3ib40215k","article-title":"Plasma Membrane Recovery Kinetics of a Microfluidic Intracellular Delivery Platform","volume":"6","author":"Sharei","year":"2014","journal-title":"Integr Biol"},{"key":"B29","doi-asserted-by":"publisher","first-page":"e0118803","DOI":"10.1371\/journal.pone.0118803","article-title":"Ex Vivo Cytosolic Delivery of Functional Macromolecules to Immune Cells","volume":"10","author":"Sharei","year":"2015","journal-title":"PloS One"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.3791\/50980","article-title":"Cell Squeezing as a Robust, Microfluidic Intracellular Delivery Platform","volume":"81","author":"Sharei","year":"2013","journal-title":"J Vis Exp"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2014.00009","article-title":"Of Macrophages and Red Blood Cells; a Complex Love Story","volume":"5","author":"de Back","year":"2014","journal-title":"Front Physiol"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1038\/nri2545","article-title":"Immunogenic and Tolerogenic Cell Death","volume":"9","author":"Green","year":"2009","journal-title":"Nat Rev Immunol"},{"key":"B33","first-page":"196","article-title":"Determination of RBC Survival in C57BL\/6 and C57BL\/6-Tg(UBC-GFP) Mice","volume":"65","author":"Dholakia","year":"2015","journal-title":"Comp Med"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2021.655393","article-title":"How Do Red Blood Cells Die","volume":"12","author":"Thiagarajan","year":"2021","journal-title":"Front Physiol"},{"key":"B35","doi-asserted-by":"publisher","first-page":"994","DOI":"10.3324\/haematol.2011.057620","article-title":"Physiologically Aged Red Blood Cells Undergo Erythrophagocytosis In Vivo But Not In Vitro","volume":"97","author":"Gottlieb","year":"2012","journal-title":"Haematologica"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1111\/trf.13497","article-title":"Erythrophagocytosis by Plasmacytoid Dendritic Cells and Monocytes is Enhanced During Inflammation","volume":"56","author":"Richards","year":"2016","journal-title":"Transfusion"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2018.04.062","article-title":"Recent Advances in the Development of Vaccines for Chronic Inflammatory Autoimmune Diseases","volume":"36","author":"Zhang","year":"2018","journal-title":"Vaccine"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1038\/nrrheum.2017.107","article-title":"Antigen-Specific Immunotherapies in Rheumatic Diseases","volume":"13","author":"Pozsgay","year":"2017","journal-title":"Nat Rev Rheumatol"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.3390\/cells10030652","article-title":"Heme Oxygenase-1 Contributes to Both the Engulfment and the Anti-Inflammatory Program of Macrophages During Efferocytosis","volume":"10","author":"Fige","year":"2021","journal-title":"Cells"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(93)90730-e","article-title":"Following a Diabetogenic T Cell From Genesis Through Pathogenesis","volume":"74","author":"Katz","year":"1993","journal-title":"Cell"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.86.20.8000","article-title":"Pancreatic Islet-Specific T-Cell Clones From Nonobese Diabetic Mice","volume":"86","author":"Haskins","year":"1989","journal-title":"Proc Natl Acad Sci"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1038\/ni.1844","article-title":"Chromogranin A is an Autoantigen in Type 1 Diabetes","volume":"11","author":"Stadinski","year":"2010","journal-title":"Nat Immunol"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1003617","article-title":"Cutting Edge: Vasostatin-1\u2013Derived Peptide ChgA29\u201342 Is an Antigenic Epitope of Diabetogenic BDC2.5 T Cells in Nonobese Diabetic Mice","volume":"186","author":"Nikoopour","year":"2011","journal-title":"J Immunol"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1093\/intimm\/dxp095","article-title":"Regulatory T Cells: How do They Suppress Immune Responses","volume":"21","author":"Sakaguchi","year":"2009","journal-title":"Int Immunol"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2008.05.009","article-title":"Regulatory T Cells and Immune Tolerance","volume":"133","author":"Sakaguchi","year":"2008","journal-title":"Cell"},{"key":"B46","doi-asserted-by":"publisher","first-page":"48","DOI":"10.4049\/jimmunol.1900127","article-title":"Nanoparticles Containing an Insulin\u2013ChgA Hybrid Peptide Protect From Transfer of Autoimmune Diabetes by Shifting the Balance Between Effector T Cells and Regulatory T Cells","volume":"203","author":"Jamison","year":"2019","journal-title":"J Immunol"},{"key":"B47","doi-asserted-by":"publisher","first-page":"595","DOI":"10.1038\/ni.1731","article-title":"The Transcription Factor T-Bet Controls Regulatory T Cell Homeostasis and Function During Type 1 Inflammation","volume":"10","author":"Koch","year":"2009","journal-title":"Nat Immunol"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2005-04-1393","article-title":"T-Bet is Required for Optimal Proinflammatory CD4+ T-Cell Trafficking","volume":"106","author":"Lord","year":"2005","journal-title":"Blood"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.96.16.9311","article-title":"Prevalent CD8+ T Cell Response Against One Peptide\/MHC Complex in Autoimmune Diabetes","volume":"96","author":"Anderson","year":"1999","journal-title":"Proc Natl Acad Sci"},{"key":"B50","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41551-021-00738-5","article-title":"Induction of Antigen-Specific Tolerance by Nanobody\u2013Antigen Adducts That Target Class-II Major Histocompatibility Complexes","volume":"5","author":"Pishesha","year":"2021","journal-title":"Nat BioMed Eng"},{"key":"B51","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1016\/b978-044452843-8\/50006-7","article-title":"Food Toxicants Analysis","author":"Lee","year":"2007","journal-title":"Part 3 Mol Biol Techniques"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1408686111","article-title":"Polymeric Synthetic Nanoparticles for the Induction of Antigen-Specific Immunological Tolerance","volume":"112","author":"Maldonado","year":"2015","journal-title":"Proc Natl Acad Sci"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaut.2017.12.010","article-title":"Tolerogenic Ag-PLG Nanoparticles Induce Tregs to Suppress Activated Diabetogenic CD4 and CD8 T Cells","volume":"89","author":"Prasad","year":"2018","journal-title":"J Autoimmun"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.00281","article-title":"Tolerogenic Nanoparticles Induce Antigen-Specific Regulatory T Cells and Provide Therapeutic Efficacy and Transferrable Tolerance Against Experimental Autoimmune Encephalomyelitis","volume":"9","author":"LaMothe","year":"2018","journal-title":"Front Immunol"},{"key":"B55","doi-asserted-by":"publisher","first-page":"1667","DOI":"10.1053\/j.gastro.2020.01.045","article-title":"Gliadin Nanoparticles Induce Immune Tolerance to Gliadin in Mouse Models of Celiac Disease","volume":"158","author":"Freitag","year":"2020","journal-title":"Gastroenterology"},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1038\/s41565-018-0273-1","article-title":"Roadmap and Strategy for Overcoming Infusion Reactions to Nanomedicines","volume":"13","author":"Szebeni","year":"2018","journal-title":"Nat Nanotechnol"},{"key":"B57","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-7352-1","article-title":"Characterization of Nanoparticles Intended for Drug Delivery","volume":"1682","author":"Neun","year":"2018","journal-title":"Methods Mol Biol"},{"key":"B58","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.173.5.2942","article-title":"CD4+CD25+ Cells Controlling a Pathogenic CD4 Response Inhibit Cytokine Differentiation, CXCR-3 Expression, and Tissue Invasion","volume":"173","author":"Sarween","year":"2004","journal-title":"J Immunol"},{"key":"B59","doi-asserted-by":"publisher","first-page":"e0126311","DOI":"10.1371\/journal.pone.0126311","article-title":"Th1-Like ICOS+ Foxp3+ Treg Cells Preferentially Express CXCR3 and Home to \u03b2-Islets During Pre-Diabetes in BDC2.5 NOD Mice","volume":"10","author":"Kornete","year":"2015","journal-title":"PloS One"},{"key":"B60","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1616710113","article-title":"Singular Role for T-BET+CXCR3+ Regulatory T Cells in Protection From Autoimmune Diabetes","volume":"113","author":"Tan","year":"2016","journal-title":"Proc Natl Acad Sci"},{"key":"B61","doi-asserted-by":"publisher","DOI":"10.1038\/s41574-019-0254-y","article-title":"The Heterogeneous Pathogenesis of Type 1 Diabetes Mellitus","volume":"15","author":"Ilonen","year":"2019","journal-title":"Nat Rev Endocrinol"},{"key":"B62","doi-asserted-by":"publisher","DOI":"10.1001\/jama.2013.6285","article-title":"Seroconversion to Multiple Islet Autoantibodies and Risk of Progression to Diabetes in Children","volume":"309","author":"Ziegler","year":"2013","journal-title":"Jama"},{"key":"B63","doi-asserted-by":"publisher","DOI":"10.2337\/dc15-1419","article-title":"Staging Presymptomatic Type 1 Diabetes: A Scientific Statement of JDRF, the Endocrine Society, and the American Diabetes Association","volume":"38","author":"Insel","year":"2015","journal-title":"Diabetes Care"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.869669\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,4]],"date-time":"2022-04-04T12:40:57Z","timestamp":1649076057000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.869669\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,4,4]]},"references-count":63,"alternative-id":["10.3389\/fimmu.2022.869669"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2022.869669","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,4,4]]},"article-number":"869669"}}