{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T16:27:29Z","timestamp":1783700849856,"version":"3.55.0"},"reference-count":37,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,7,15]],"date-time":"2022-07-15T00:00:00Z","timestamp":1657843200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>Regulatory T cells (Tregs) are normally born in the thymus and activated in secondary lymphoid tissues to suppress immune responses in the lymph node and at sites of inflammation. Tregs are also resident in various tissues or accumulate in damaged tissues, which are now called tissue Tregs, and contribute to homeostasis and tissue repair by interacting with non-immune cells. We have shown that Tregs accumulate in the brain during the chronic phase in a mouse cerebral infarction model, and these Tregs acquire the characteristic properties of brain Tregs and contribute to the recovery of neurological damage by interacting with astrocytes. However, the mechanism of tissue Treg development is not fully understood. We developed a culture method that confers brain Treg characteristics <jats:italic>in vitro<\/jats:italic>. Naive Tregs from the spleen were activated and efficiently amplified by T-cell receptor (TCR) stimulation in the presence of primary astrocytes. Furthermore, adding IL-33 and serotonin could confer part of the properties of brain Tregs, such as ST2, peroxisome proliferator-activated receptor \u03b3 (PPAR\u03b3), and serotonin receptor 7 (Htr7) expression. Transcriptome analysis revealed that <jats:italic>in vitro<\/jats:italic> generated brain Treg-like Tregs (induced brain Tregs; iB-Tregs) showed similar gene expression patterns as those in <jats:italic>in vivo<\/jats:italic> brain Tregs, although they were not identical. Furthermore, in Parkinson\u2019s disease models, in which T cells have been shown to be involved in disease progression, iB-Tregs infiltrated into the brain more readily and ameliorated pathological symptoms more effectively than splenic Tregs. These data indicate that iB-Tregs contribute to our understanding of brain Treg development and could also be therapeutic for inflammatory brain diseases.<\/jats:p>","DOI":"10.3389\/fimmu.2022.960036","type":"journal-article","created":{"date-parts":[[2022,7,15]],"date-time":"2022-07-15T11:21:18Z","timestamp":1657884078000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":20,"title":["In Vitro Generation of Brain Regulatory T Cells by Co-culturing With Astrocytes"],"prefix":"10.3389","volume":"13","author":[{"given":"Shinichi","family":"Yamamoto","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ako","family":"Matsui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masaki","family":"Ohyagi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chie","family":"Kikutake","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshihiro","family":"Harada","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mana","family":"Iizuka-Koga","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikita","family":"Suyama","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akihiko","family":"Yoshimura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minako","family":"Ito","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,7,15]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1093\/intimm\/dxz031","article-title":"Tissue Regulatory T Cells and Neural Repair","volume":"31","author":"Ito","year":"2019","journal-title":"Int Immunol"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-immunol-032712-095948","article-title":"Tissue Tregs","volume":"34","author":"Panduro","year":"2016","journal-title":"Annu Rev Immunol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.08.021","article-title":"A Distinct Function of Regulatory T Cells in Tissue Protection","volume":"162","author":"Arpaia","year":"2015","journal-title":"Cell"},{"key":"B4","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1016\/j.immuni.2019.01.001","article-title":"Single-Cell Transcriptomics of Regulatory T Cells Reveals Trajectories of Tissue Adaptation","volume":"50","author":"Miragaia","year":"2019","journal-title":"Immunity"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1126\/sciimmunol.aat5861","article-title":"Molecular Diversification of Regulatory T Cells in Nonlymphoid Tissues","volume":"3","author":"DiSpirito","year":"2018","journal-title":"Sci Immunol"},{"key":"B6","doi-asserted-by":"publisher","first-page":"285","DOI":"10.1016\/j.cell.2018.05.004","article-title":"TCR Transgenic Mice Reveal Stepwise, Multi-Site Acquisition of the Distinctive Fat-Treg Phenotype","volume":"174","author":"Li","year":"2018","journal-title":"Cell"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2013.10.054","article-title":"A Special Population of Regulatory T Cells Potentiates Muscle Repair","volume":"155","author":"Burzyn","year":"2013","journal-title":"Cell"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2016.01.009","article-title":"Poor Repair of Skeletal Muscle in Aging Mice Reflects a Defect in Local, Interleukin-33-Dependent Accumulation of Regulatory T Cells","volume":"44","author":"Kuswanto","year":"2016","journal-title":"Immunity"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1038\/s41423-020-00547-x","article-title":"Kidney GATA3(+) Regulatory T Cells Play Roles in the Convalescence Stage After Antibody-Mediated Renal Injury","volume":"18","author":"Sakai","year":"2021","journal-title":"Cell Mol Immunol"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0824-5","article-title":"Brain Regulatory T Cells Suppress Astrogliosis and Potentiate Neurological Recovery","volume":"565","author":"Ito","year":"2019","journal-title":"Nature"},{"key":"B11","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.immuni.2021.04.022","article-title":"Treg Cell-Derived Osteopontin Promotes Microglia-Mediated White Matter Repair After Ischemic Stroke","volume":"54","author":"Ligen Shi","year":"2021","journal-title":"Immunity"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0901852","article-title":"Regulatory T Cells Attenuate Th17 Cell-Mediated Nigrostriatal Dopaminergic Neurodegeneration in a Model of Parkinson\u2019s Disease","volume":"184","author":"Reynolds","year":"2010","journal-title":"J Immunol"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2019.00421","article-title":"A Synthetic Agonist to Vasoactive Intestinal Peptide Receptor-2 Induces Regulatory T Cell Neuroprotective Activities in Models of Parkinson\u2019s Disease","volume":"13","author":"Mosley","year":"2019","journal-title":"Front Cell Neurosci"},{"key":"B14","doi-asserted-by":"publisher","first-page":"268","DOI":"10.1016\/j.immuni.2017.07.008","article-title":"Analyses of a Mutant Foxp3 Allele Reveal BATF as a Critical Transcription Factor in the Differentiation and Accumulation of Tissue Regulatory T Cells","volume":"47","author":"Hayatsu","year":"2017","journal-title":"Immunity"},{"key":"B15","doi-asserted-by":"publisher","first-page":"100954","DOI":"10.1016\/j.xpro.2021.100954","article-title":"Isolation and Culturing of Primary Mouse Astrocytes for the Analysis of Focal Adhesion Dynamics","volume":"2","author":"G\u00fcler","year":"2021","journal-title":"STAR Protoc"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1016\/j.neulet.2010.01.007","article-title":"Close Association of P38 and JNK With Plasminogen-Dependent Upregulation of PAI-1 in Rat Astrocytes In Vitro","volume":"471","author":"Nakajima","year":"2010","journal-title":"Neurosci Lett"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbadis.2007.12.002","article-title":"Microglia and Astroglia Prevent Oxidative Stress-Induced Neuronal Cell Death: Implications for Aceruloplasminemia","volume":"1782","author":"Oshiro","year":"2008","journal-title":"Biochim Biophys Acta"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1002\/eji.200535430","article-title":"Vasoactive Intestinal Peptide Induces Regulatory T Cells During Experimental Autoimmune Encephalomyelitis","volume":"36","author":"Fernandez-Martin","year":"2006","journal-title":"Eur J Immunol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1007\/s00401-016-1631-4","article-title":"Multiple Sclerosis: Experimental Models and Reality","volume":"133","author":"Lassmann","year":"2017","journal-title":"Acta neuropathologica"},{"key":"B20","doi-asserted-by":"publisher","first-page":"300060521998471","DOI":"10.1177\/0300060521998471","article-title":"Imbalance Between T Helper 1 and Regulatory T Cells Plays a Detrimental Role in Experimental Parkinson\u2019s Disease in Mice","volume":"49","author":"Li","year":"2021","journal-title":"J Int Med Res"},{"key":"B21","doi-asserted-by":"publisher","first-page":"104575","DOI":"10.1016\/j.nbd.2019.104575","article-title":"RANTES-Induced Invasion of Th17 Cells Into Substantia Nigra Potentiates Dopaminergic Cell Loss in MPTP Mouse Model of Parkinson\u2019s Disease","volume":"132","author":"Dutta","year":"2019","journal-title":"Neurobiol Dis"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.3389\/fnagi.2018.00179","article-title":"Comparison of Administration Routes on the Protective Effects of Bee Venom Phospholipase A2 in a Mouse Model of Parkinson\u2019s Disease","volume":"10","author":"Baek","year":"2018","journal-title":"Front Aging Neurosci"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1922559117","article-title":"Neuronal, Stromal, and T-Regulatory Cell Crosstalk in Murine Skeletal Muscle","volume":"117","author":"Wang","year":"2020","journal-title":"Proc Natl Acad Sci USA"},{"key":"B24","doi-asserted-by":"publisher","first-page":"4011","DOI":"10.1038\/s41467-020-17750-z","article-title":"Blockade of the AHR Restricts a Treg-Macrophage Suppressive Axis Induced by L-Kynurenine","volume":"11","author":"Campesato","year":"2020","journal-title":"Nat Commun"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1126\/sciimmunol.abi4613","article-title":"Transforming Growth Factor-\u03b21 in Regulatory T Cell Biology","volume":"7","author":"Moreau","year":"2022","journal-title":"Sci Immunol"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.198382","article-title":"Multiple Retinol and Retinal Dehydrogenases Catalyze All-Trans-Retinoic Acid Biosynthesis in Astrocytes","volume":"286","author":"Wang","year":"2011","journal-title":"J Biol Chem"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1038\/nature12428","article-title":"Stability and Function of Regulatory T Cells Is Maintained by a Neuropilin-1-Semaphorin-4a Axis","volume":"501","author":"Delgoffe","year":"2013","journal-title":"Nature"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.3389\/fnsys.2021.682990","article-title":"Increasing Serotonin to Reduce Parkinsonian Tremor","volume":"15","author":"Caligiore","year":"2021","journal-title":"Front Syst Neurosci"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1038\/nm.1927","article-title":"Regulatory T Cells are Key Cerebroprotective Immunomodulators in Acute Experimental Stroke","volume":"15","author":"Liesz","year":"2009","journal-title":"Nat Med"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1801330","article-title":"Innate Control of Tissue-Reparative Human Regulatory T Cells","volume":"202","author":"Lam","year":"2019","journal-title":"J Immunol"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1038\/ni.2520","article-title":"Nr4a Receptors are Essential for Thymic Regulatory T Cell Development and Immune Homeostasis","volume":"14","author":"Sekiya","year":"2013","journal-title":"Nat Immunol"},{"key":"B32","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1084\/jem.20061852","article-title":"Selective Depletion of Foxp3+ Regulatory T Cells Induces a Scurfy-Like Disease","volume":"204","author":"Lahl","year":"2007","journal-title":"J Exp Med"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.763647","article-title":"Single-Cell Analysis Revealed the Role of CD8(+) Effector T Cells in Preventing Cardioprotective Macrophage Differentiation in the Early Phase of Heart Failure","volume":"12","author":"Komai","year":"2021","journal-title":"Front Immunol"},{"key":"B34","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1016\/j.brainresbull.2020.08.015","article-title":"Echinacoside Protects Dopaminergic Neurons by Inhibiting NLRP3\/Caspase-1\/IL-1\u03b2 Signaling Pathway in MPTP-Induced Parkinson\u2019s Disease Model","volume":"164","author":"Gao","year":"2020","journal-title":"Brain Res Bull"},{"key":"B35","doi-asserted-by":"publisher","first-page":"7360","DOI":"10.1038\/ncomms8360","article-title":"Bruton\u2019s Tyrosine Kinase Is Essential for NLRP3 Inflammasome Activation and Contributes to Ischaemic Brain Injury","volume":"6","author":"Ito","year":"2015","journal-title":"Nat Commun"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2011.02.021","article-title":"Transcription Factor Smad-Independent T Helper 17 Cell Induction by Transforming-Growth Factor-\u03b2 Is Mediated by Suppression of Eomesodermin","volume":"34","author":"Ichiyama","year":"2011","journal-title":"Immunity"},{"key":"B37","doi-asserted-by":"publisher","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated Estimation of Fold Change and Dispersion for RNA-Seq Data With Deseq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.960036\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,7,15]],"date-time":"2022-07-15T11:21:20Z","timestamp":1657884080000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.960036\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,15]]},"references-count":37,"alternative-id":["10.3389\/fimmu.2022.960036"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2022.960036","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,15]]},"article-number":"960036"}}