^{Joint Rheumatology Program, Clinical Immunology-Rheumatology Unit, National and Kapodistrian University of Athens Medical School, Hippokration General Hospital, Athens, Greece.}

^{Joint Rheumatology Program, 4th Department of Medicine, Attikon University Hospital, National and Kapodistrian University of Athens Medical School, Athens, Greece.}

^{Department of Clinical Pathobiochemistry, Institute for Clinical Chemistry and Laboratory Medicine and Department of Internal Medicine, University of Dresden, Dresden, Germany.}

^{Kimmel Center for Biology and Medicine at the Skirball Institute, New York, New York, USA.}

^{Departments of Microbiology and Medicine, New York University School of Medicine, New York, New York, USA.}

^{Corresponding author.}

Address correspondence to: Panayotis Verginis, Center of Clinical, Experimental Surgery, and Translational Research, Biomedical Research Foundation, Academy of Athens, 115 27 Athens, Greece. Phone: 30.210.6597516; Email: rg.ymedacaoib@sinigrevp.

Themis Alissafi: moc.liamg@ifassilat; Aggelos Banos: rg.ymedacaoib@sonaba; Louis Boon: moc.sorecoib@noob.l; Tim Sparwasser: ed.revonnah-hm@miT.ressawrapS; Alessandra Ghigo: ti.otinu@ogihg.ardnassela; Kajsa Wing: es.ik@gniW.asjaK; Dimitrios Vassilopoulos: rg.aou.dem@polissavd; Dimitrios Boumpas: rg.cou@dsapmuob; Triantafyllos Chavakis: ed.nedserd-mukinilkinu@sikavahc.sollyfatnairt; Panayotis Verginis: rg.ymedacaoib@sinigrevpAddress correspondence to: Panayotis Verginis, Center of Clinical, Experimental Surgery, and Translational Research, Biomedical Research Foundation, Academy of Athens, 115 27 Athens, Greece. Phone: 30.210.6597516; Email: rg.ymedacaoib@sinigrevp.

Received 2016 Dec 1; Accepted 2017 Apr 7.

Abstract

Design of efficacious Treg-based therapies and establishment of clinical tolerance in autoimmune diseases have proven to be challenging. The clinical implementation of Treg immunotherapy has been hampered by various impediments related to the stability and isolation procedures of Tregs as well as the specific in vivo targets of Treg modalities. Herein, we have demonstrated that Foxp3^{Tregs potently suppress autoimmune responses in vivo through inhibition of the autophagic machinery in DCs in a cytotoxic T-lymphocyte–associated protein 4–dependent (CTLA4-dependent) manner. Autophagy-deficient DCs exhibited reduced immunogenic potential and failed to prime autoantigen-specific CD4}^{T cells to mediate autoimmunity. Mechanistically, CTLA4 binding promoted activation of the PI3K/Akt/mTOR axis and FoxO1 nuclear exclusion in DCs, leading to decreased transcription of the autophagy component microtubule-associated protein 1 light chain 3β (}Lc3b). Human DCs treated with CTLA4-Ig, a fusion protein composed of the Fc region of IgG1 and the extracellular domain of CTLA4 (also known as abatacept, marketed as Orencia), demonstrated reduced levels of autophagosome formation, while DCs from CTLA4-Ig–treated rheumatoid arthritis patients displayed diminished LC3B transcripts. Collectively, our data identify the canonical autophagy pathway in DCs as a molecular target of Foxp3^{Treg–mediated suppression that leads to amelioration of autoimmune responses. These findings may pave the way for the development of therapeutic protocols that exploit Tregs for the treatment of autoimmunity as well as diseases in which disturbed tolerance is a common denominator.}

Keywords: Autoimmunity, Immunology

Abstract

Acknowledgments

We would like to thank Katerina Hatziioannou, Marianna Ioannou, Katerina Girtzimanaki, Eirini Kirmizi, and Nikos Paschalidis for assisting with experiments; George Chamilos for providing technical advice; Anastasia Apostolidou and Arianna Gavriil for cell sorting; nurses Giota Rapsomaniki and Kalliopi Klavdianou for patient care; Ismini Kloukina for histology preparations; Stamatis Pagakis and Eleni Rigana for providing assistance with confocal microscopy and quantification of confocal images; Watanebe Masashi for assisting with the B7.1 B7.2 double-knockout samples; Katrin Klocke for assisting with the Rosa26-Cre^Ctla4^fl/fl mice; Emilio Hirsch and Alessandra Chigo for providing the Pik3cg^–/– mice; and Christos Tsatsanis for providing the Akt1^–/– mice. This work was supported by grants from the Greek General Secretariat of Research and Technology (Aristeia II 3468 to PV), European Union project Innovative Medicine Initiative 6 (BeTheCure, contract number 115142-2, to PV and DB). TC is supported by the Deutsche Forschungsgemeinschaft (CH279/5-1 and SFB655). TA is supported by the European Commission FP7 programme Translational Potential (TransPOT; EC contract number 285948) and IKY Fellowships of Excellence for Postgraduate Studies in Greece — Siemens Programme. KC is supported by NIH grants ({"type":"entrez-nucleotide","attrs":{"text":"DK103788","term_id":"187403994","term_text":"DK103788"}}DK103788, {"type":"entrez-nucleotide","attrs":{"text":"DK093668","term_id":"187545272","term_text":"DK093668"}}DK093668, {"type":"entrez-nucleotide","attrs":{"text":"HL123340","term_id":"1051707559","term_text":"HL123340"}}HL123340) and is a Burroughs Welcome Fund Investigator in the Pathogenesis of Infectious Diseases.

Acknowledgments

Footnotes

Conflict of interest: The authors have declared that no conflict of interest exists.

Reference information:J Clin Invest. 2017;127(7):2789–2804.https://doi.org/10.1172/JCI92079.

Footnotes

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