Regulatory T cells (Tregs) are indispensable for maintaining self-tolerance by suppressing conventional T cells. On the other hand, Tregs promote tumor growth by inhibiting anticancer immunity. In this study, we identified that Tregs increase the quorum of self-reactive CD8+ T cells required for the induction of experimental autoimmune diabetes in mice. Their major suppression mechanism is limiting available IL-2, an essential T-cell cytokine. Specifically, Tregs inhibit the formation of a previously uncharacterized subset of antigen-stimulated KLRK1+ IL-7R+ (KILR) CD8+ effector T cells, which are distinct from conventional effector CD8+ T cells. KILR CD8+ T cells show superior cell-killing abilities in vivo. The administration of agonistic IL-2 immunocomplexes phenocopies the absence of Tregs, i.e., it induces KILR CD8+ T cells, promotes autoimmunity, and enhances antitumor responses in mice. Counterparts of KILR CD8+ T cells were found in the human blood, revealing them as a potential target for immunotherapy.

As well as protecting us from invading pathogens, like bacteria or viruses, our immune system can also identify dangerous cells of our own that may cause the body harm, such as cancer cells. Once detected, a population of immune cells called cytotoxic T cells launch into action to kill the potentially harmful cell. However, sometimes the immune system makes mistakes and attacks healthy cells which it misidentifies as being dangerous, leading to autoimmune diseases. Special immune cells called T regulatory lymphocytes, or ‘Tregs’, can suppress the activity of cytotoxic T cells, preventing them from hurting the body’s own cells. While this can have a positive impact and reduce the effects of autoimmunity, Tregs can also make the immune system less responsive to cancer cells and allow tumors to grow. But how Tregs alter the behavior of cytotoxic T cells during autoimmune diseases and cancer is poorly understood. While multiple mechanisms have been proposed, none of these have been tested in living animal models of these diseases. To address this, Tsyklauri et al. studied Tregs in laboratory mice which had been modified to have autoimmune diabetes, which is when the body attacks the cells responsible for producing insulin. The experiments revealed that Tregs take up a critical signaling molecule called IL-2 which cytotoxic T cells need to survive and multiply. As a result, there is less IL-2 molecules available in the environment, inhibiting the cytotoxic T cells’ activity. Furthermore, if Tregs are absent and there is an excess of IL-2, this causes cytotoxic T cells to transition into a previously unknown subset of T cells with superior killing abilities. Tsyklauri et al. were able to replicate these findings in two different groups of laboratory mice which had been modified to have cancer. This suggests that Tregs suppress the immune response to cancer cells and prevent autoimmunity using the same mechanism. In the future, this work could help researchers to develop therapies that alter the behavior of cytotoxic T cells and/or Tregs to either counteract autoimmune diseases, or help the body fight off cancer.

Department of Biomedicine University Hospital of Basel Basel Switzerland

Faculty of Science Charles University Prague Czech Republic

Institute of Microbiology of the Czech Academy of Sciences Prague Czech Republic

Laboratory of Adaptive Immunity Institute of Molecular Genetics of the Czech Academy of Sciences Prague Czech Republic

Před aktualizací

doi: 10.1101/2021.11.10.467495 PubMed

Zobrazit více v PubMed

Akkaya B, Oya Y, Akkaya M, Al Souz JA, Holstein AH, Kabat J, Kamenyeva O, Dorward D, Glass D, Shevach EM. Regulatory T cells perform antigen specific suppression by depleting cognate peptide-MHC class II via trogocytosis. The Journal of Immunology. 2019;202:57. doi: 10.4049/jimmunol.202.Supp.57.17. PubMed DOI PMC

Andreatta M, Corria-Osorio J, Müller S, Cubas R, Coukos G, Carmona SJ. Interpretation of T cell states from single-cell transcriptomics data using reference atlases. Nature Communications. 2021;12:2965. doi: 10.1038/s41467-021-23324-4. PubMed DOI PMC

Bais AS, Kostka D. Scds: computational annotation of doublets in single-cell RNA sequencing data. Bioinformatics. 2020;36:1150–1158. doi: 10.1093/bioinformatics/btz698. PubMed DOI PMC

Barnden MJ, Allison J, Heath WR, Carbone FR. Defective TCR expression in transgenic mice constructed using cDNA-based alpha- and beta-chain genes under the control of heterologous regulatory elements. Immunology and Cell Biology. 1998;76:34–40. doi: 10.1046/j.1440-1711.1998.00709.x. PubMed DOI

Barsoumian HB, Yolcu ES, Shirwan H. 4-1Bb signaling in conventional T cells drives IL-2 production that overcomes CD4+CD25+FoxP3+ T regulatory cell suppression. PLOS ONE. 2016;11:e0153088. doi: 10.1371/journal.pone.0153088. PubMed DOI PMC

Bennett CL, Christie J, Ramsdell F, Brunkow ME, Ferguson PJ, Whitesell L, Kelly TE, Saulsbury FT, Chance PF, Ochs HD. The immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) is caused by mutations of FOXP3. Nature Genetics. 2001;27:20–21. doi: 10.1038/83713. PubMed DOI

Bosch AJT, Bolinger B, Keck S, Stepanek O, Ozga AJ, Galati-Fournier V, Stein JV, Palmer E. A minimum number of autoimmune T cells to induce autoimmunity? Cellular Immunology. 2017;316:21–31. doi: 10.1016/j.cellimm.2017.03.002. PubMed DOI

Boyman O, Kovar M, Rubinstein MP, Surh CD, Sprent J. Selective stimulation of T cell subsets with antibody-cytokine immune complexes. Science. 2006;311:1924–1927. doi: 10.1126/science.1122927. PubMed DOI

Brunkow ME, Jeffery EW, Hjerrild KA, Paeper B, Clark LB, Yasayko SA, Wilkinson JE, Galas D, Ziegler SF, Ramsdell F. Disruption of a new forkhead/winged-helix protein, scurfin, results in the fatal lymphoproliferative disorder of the scurfy mouse. Nature Genetics. 2001;27:68–73. doi: 10.1038/83784. PubMed DOI

Chinen T, Kannan AK, Levine AG, Fan X, Klein U, Zheng Y, Gasteiger G, Feng Y, Fontenot JD, Rudensky AY. An essential role for the IL-2 receptor in Treg cell function. Nature Immunology. 2016;17:1322–1333. doi: 10.1038/ni.3540. PubMed DOI PMC

Dong S, Hiam-Galvez KJ, Mowery CT, Herold KC, Gitelman SE, Esensten JH, Liu W, Lares AP, Leinbach AS, Lee M, Nguyen V, Tamaki SJ, Tamaki W, Tamaki CM, Mehdizadeh M, Putnam AL, Spitzer MH, Ye CJ, Tang Q, Bluestone JA. The effect of low-dose IL-2 and treg adoptive cell therapy in patients with type 1 diabetes. JCI Insight. 2021;6:18. doi: 10.1172/jci.insight.147474. PubMed DOI PMC

Green EA, Gorelik L, McGregor CM, Tran EH, Flavell RA. Cd4+Cd25+ T regulatory cells control anti-islet CD8+ T cells through TGF-beta-TGF-beta receptor interactions in type 1 diabetes. PNAS. 2003;100:10878–10883. doi: 10.1073/pnas.1834400100. PubMed DOI PMC

Haberman Y, Tickle TL, Dexheimer PJ, Kim M-O, Tang D, Karns R, Baldassano RN, Noe JD, Rosh J, Markowitz J, Heyman MB, Griffiths AM, Crandall WV, Mack DR, Baker SS, Huttenhower C, Keljo DJ, Hyams JS, Kugathasan S, Walters TD, Aronow B, Xavier RJ, Gevers D, Denson LA. Pediatric crohn disease patients exhibit specific ileal transcriptome and microbiome signature. The Journal of Clinical Investigation. 2014;124:3617–3633. doi: 10.1172/JCI75436. PubMed DOI PMC

Hahsler M. Dbscan: fast density-based clustering with R. Journal of Statistical Software. 2019;91:1–30. doi: 10.18637/jss.v091.i01. DOI

Howe KL, Achuthan P, Allen J, Allen J, Alvarez-Jarreta J, Amode MR, Armean IM, Azov AG, Bennett R, Bhai J, Billis K, Boddu S, Charkhchi M, Cummins C, Da Rin Fioretto L, Davidson C, Dodiya K, El Houdaigui B, Fatima R, Gall A, Garcia Giron C, Grego T, Guijarro-Clarke C, Haggerty L, Hemrom A, Hourlier T, Izuogu OG, Juettemann T, Kaikala V, Kay M, Lavidas I, Le T, Lemos D, Gonzalez Martinez J, Marugán JC, Maurel T, McMahon AC, Mohanan S, Moore B, Muffato M, Oheh DN, Paraschas D, Parker A, Parton A, Prosovetskaia I, Sakthivel MP, Salam AIA, Schmitt BM, Schuilenburg H, Sheppard D, Steed E, Szpak M, Szuba M, Taylor K, Thormann A, Threadgold G, Walts B, Winterbottom A, Chakiachvili M, Chaubal A, De Silva N, Flint B, Frankish A, Hunt SE, IIsley GR, Langridge N, Loveland JE, Martin FJ, Mudge JM, Morales J, Perry E, Ruffier M, Tate J, Thybert D, Trevanion SJ, Cunningham F, Yates AD, Zerbino DR, Flicek P. Ensembl 2021. Nucleic Acids Research. 2021;49:D884–D891. doi: 10.1093/nar/gkaa942. PubMed DOI PMC

Jang Y, Gerbec ZJ, Won T, Choi B, Podsiad A, B Moore B, Malarkannan S, Laouar Y. Cutting edge: check your mice-A point mutation in the ncr1 locus identified in CD45.1 congenic mice with consequences in mouse susceptibility to infection. Journal of Immunology. 2018;200:1982–1987. doi: 10.4049/jimmunol.1701676. PubMed DOI PMC

Josefowicz SZ, Rudensky A. Control of regulatory T cell lineage commitment and maintenance. Immunity. 2009;30:616–625. doi: 10.1016/j.immuni.2009.04.009. PubMed DOI PMC

Joshi NS, Cui W, Chandele A, Lee HK, Urso DR, Hagman J, Gapin L, Kaech SM. Inflammation directs memory precursor and short-lived effector CD8 (+) T cell fates via the graded expression of T-bet transcription factor. Immunity. 2007;27:281–295. doi: 10.1016/j.immuni.2007.07.010. PubMed DOI PMC

Kaech SM, Tan JT, Wherry EJ, Konieczny BT, Surh CD, Ahmed R. Selective expression of the interleukin 7 receptor identifies effector CD8 T cells that give rise to long-lived memory cells. Nature Immunology. 2003;4:1191–1198. doi: 10.1038/ni1009. PubMed DOI

Kalia V, Penny LA, Yuzefpolskiy Y, Baumann FM, Sarkar S. Quiescence of memory CD8 (+) T cells is mediated by regulatory T cells through inhibitory receptor CTLA-4. Immunity. 2015;42:1116–1129. doi: 10.1016/j.immuni.2015.05.023. PubMed DOI

Kastenmuller W, Gasteiger G, Subramanian N, Sparwasser T, Busch DH, Belkaid Y, Drexler I, Germain RN. Regulatory T cells selectively control CD8+ T cell effector pool size via IL-2 restriction. Journal of Immunology. 2011;187:3186–3197. doi: 10.4049/jimmunol.1101649. PubMed DOI PMC

Kim JM, Rasmussen JP, Rudensky AY. Regulatory T cells prevent catastrophic autoimmunity throughout the lifespan of mice. Nature Immunology. 2007;8:191–197. doi: 10.1038/ni1428. PubMed DOI

Kim M, Ouyang W, Liao W, Zhang M, Li M. Murine in vivo CD8+ T cell killing assay. BIO-PROTOCOL. 2014;4:13. doi: 10.21769/BioProtoc.1172. PubMed DOI PMC

Kim R, Kin T. Current and future therapies for immunogenic cell death and related molecules to potentially cure primary breast cancer. Cancers. 2021;13:19. doi: 10.3390/cancers13194756. PubMed DOI PMC

King CG, Koehli S, Hausmann B, Schmaler M, Zehn D, Palmer E. T cell affinity regulates asymmetric division, effector cell differentiation, and tissue pathology. Immunity. 2012;37:709–720. doi: 10.1016/j.immuni.2012.06.021. PubMed DOI PMC

Korotkevich G. Fast Gene Set Enrichment Analysis. bioRxiv. 2021 doi: 10.1101/060012. DOI

Kovanen PE, Young L, Al-Shami A, Rovella V, Pise-Masison CA, Radonovich MF, Powell J, Fu J, Brady JN, Munson PJ, Leonard WJ. Global analysis of IL-2 target genes: identification of chromosomal clusters of expressed genes. International Immunology. 2005;17:1009–1021. doi: 10.1093/intimm/dxh283. PubMed DOI

Kralova J, Glatzova D, Borna S, Brdicka T. Expression of fluorescent fusion proteins in murine bone marrow-derived dendritic cells and macrophages. Journal of Visualized Experiments. 2018;2018:140. doi: 10.3791/58081-v. PubMed DOI

Kurts C, Miller JF, Subramaniam RM, Carbone FR, Heath WR. Major histocompatibility complex class I-restricted cross-presentation is biased towards high dose antigens and those released during cellular destruction. The Journal of Experimental Medicine. 1998;188:409–414. doi: 10.1084/jem.188.2.409. PubMed DOI PMC

Lab of Adaptive Immunity Supereffectors_scRNAseq. swh:1:rev:7b0dec9507dd45ab4bb0619912b240f512c7798fSoftware Heritage. 2023a https://archive.softwareheritage.org/swh:1:dir:a24b54a92579b7b10530c1119bc81ae22e433076;origin=https://github.com/Lab-of-Adaptive-Immunity/Supereffectors_scRNAseq;visit=swh:1:snp:2315a288b71fbc93e22b85ae7924b45146e00ab9;anchor=swh:1:rev:7b0dec9507dd45ab4bb0619912b240f512c7798f

Lab of Adaptive Immunity HS-CD8-atlas. swh:1:rev:9b31b54fff516eba2a3ddb66449eb100db16521bSoftware Heritage. 2023b https://archive.softwareheritage.org/swh:1:dir:1890b7a68107682d810bbd65ce12920c1a8747de;origin=https://github.com/Lab-of-Adaptive-Immunity/HS-CD8-Atlas;visit=swh:1:snp:f6b1a4ee7d7e0ba3e4e153f996f0a34d4f318945;anchor=swh:1:rev:9b31b54fff516eba2a3ddb66449eb100db16521b

Lahl K, Loddenkemper C, Drouin C, Freyer J, Arnason J, Eberl G, Hamann A, Wagner H, Huehn J, Sparwasser T. Selective depletion of Foxp3+ regulatory T cells induces a scurfy-like disease. The Journal of Experimental Medicine. 2007;204:57–63. doi: 10.1084/jem.20061852. PubMed DOI PMC

Laidlaw BJ, Cui W, Amezquita RA, Gray SM, Guan T, Lu Y, Kobayashi Y, Flavell RA, Kleinstein SH, Craft J, Kaech SM. Production of IL-10 by CD4 (+) regulatory T cells during the resolution of infection promotes the maturation of memory CD8 (+) T cells. Nature Immunology. 2015;16:871–879. doi: 10.1038/ni.3224. PubMed DOI PMC

Lee SJ, Rossi RJ, Lee SK, Croft M, Kwon BS, Mittler RS, Vella AT. Cd134 costimulation couples the CD137 pathway to induce production of supereffector CD8 T cells that become IL-7 dependent. Journal of Immunology. 2007;179:2203–2214. doi: 10.4049/jimmunol.179.4.2203. PubMed DOI

Lin J-X, Li P, Liu D, Jin HT, He J, Ata Ur Rasheed M, Rochman Y, Wang L, Cui K, Liu C, Kelsall BL, Ahmed R, Leonard WJ. Critical role of STAT5 transcription factor tetramerization for cytokine responses and normal immune function. Immunity. 2012;36:586–599. doi: 10.1016/j.immuni.2012.02.017. PubMed DOI PMC

Liu R, Zhou Q, La Cava A, Campagnolo DI, Van Kaer L, Shi F-D. Expansion of regulatory T cells via IL-2/anti-IL-2 mAb complexes suppresses experimental myasthenia. European Journal of Immunology. 2010;40:1577–1589. doi: 10.1002/eji.200939792. PubMed DOI PMC

Marchingo JM, Kan A, Sutherland RM, Duffy KR, Wellard CJ, Belz GT, Lew AM, Dowling MR, Heinzel S, Hodgkin PD. Antigen affinity, costimulation, and cytokine inputs sum linearly to amplify T cell expansion. Science. 2014;346:1123–1127. doi: 10.1126/science.1260044. PubMed DOI

McNally A, Hill GR, Sparwasser T, Thomas R, Steptoe RJ. Cd4+Cd25+ regulatory T cells control CD8+ T-cell effector differentiation by modulating IL-2 homeostasis. PNAS. 2011;108:7529–7534. doi: 10.1073/pnas.1103782108. PubMed DOI PMC

Obeid M, Tesniere A, Ghiringhelli F, Fimia GM, Apetoh L, Perfettini J-L, Castedo M, Mignot G, Panaretakis T, Casares N, Métivier D, Larochette N, van Endert P, Ciccosanti F, Piacentini M, Zitvogel L, Kroemer G. Calreticulin exposure dictates the immunogenicity of cancer cell death. Nature Medicine. 2007;13:54–61. doi: 10.1038/nm1523. PubMed DOI

Oh HS, Choi BK, Kim YH, Lee DG, Hwang S, Lee MJ, Park SH, Bae Y-S, Kwon BS. 4-1Bb signaling enhances primary and secondary population expansion of CD8+ T cells by maximizing autocrine IL-2/IL-2 receptor signaling. PLOS ONE. 2015;10:e0126765. doi: 10.1371/journal.pone.0126765. PubMed DOI PMC

Pace L, Tempez A, Arnold-Schrauf C, Lemaitre F, Bousso P, Fetler L, Sparwasser T, Amigorena S. Regulatory T cells increase the avidity of primary CD8+ T cell responses and promote memory. Science. 2012;338:532–536. doi: 10.1126/science.1227049. PubMed DOI

Palmer E, Drobek A, Stepanek O. Opposing effects of actin signaling and LFA-1 on establishing the affinity threshold for inducing effector T-cell responses in mice. European Journal of Immunology. 2016;46:1887–1901. doi: 10.1002/eji.201545909. PubMed DOI

Polhill T, Zhang GY, Hu M, Sawyer A, Zhou JJ, Saito M, Webster KE, Wang Y, Wang Y, Grey ST, Sprent J, Harris DCH, Alexander SI, Wang YM. IL-2/IL-2ab complexes induce regulatory T cell expansion and protect against proteinuric CKD. Journal of the American Society of Nephrology. 2012;23:1303–1308. doi: 10.1681/ASN.2011111130. PubMed DOI PMC

Prajapati K, Perez C, Rojas LBP, Burke B, Guevara-Patino JA. Functions of NKG2D in CD8+ T cells: an opportunity for immunotherapy. Cellular & Molecular Immunology. 2018;15:470–479. doi: 10.1038/cmi.2017.161. PubMed DOI PMC

Schildknecht A, Brauer S, Brenner C, Lahl K, Schild H, Sparwasser T, Probst HC, van den Broek M. Foxp3+ regulatory T cells essentially contribute to peripheral CD8+ T-cell tolerance induced by steady-state dendritic cells. PNAS. 2010;107:199–203. doi: 10.1073/pnas.0910620107. PubMed DOI PMC

Schorer M, Lambert K, Rakebrandt N, Rost F, Kao KC, Yermanos A, Spörri R, Oderbolz J, Raeber ME, Keller CW, Lünemann JD, Rogler G, Boyman O, Oxenius A, Joller N. Rapid expansion of treg cells protects from collateral colitis following a viral trigger. Nature Communications. 2020;11:1522. doi: 10.1038/s41467-020-15309-6. PubMed DOI PMC

Sereti I, Gea-Banacloche J, Kan MY, Hallahan CW, Lane HC. Interleukin 2 leads to dose-dependent expression of the alpha chain of the IL-2 receptor on CD25-negative T lymphocytes in the absence of exogenous antigenic stimulation. Clinical Immunology. 2000;97:266–276. doi: 10.1006/clim.2000.4929. PubMed DOI

Shinkai Y, Rathbun G, Lam KP, Oltz EM, Stewart V, Mendelsohn M, Charron J, Datta M, Young F, Stall AM. RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V (D) J rearrangement. Cell. 1992;68:855–867. doi: 10.1016/0092-8674(92)90029-c. PubMed DOI

Slavin S, Strober S. Spontaneous murine B-cell leukaemia. Nature. 1978;272:624–626. doi: 10.1038/272624a0. PubMed DOI

Spangler JB, Tomala J, Luca VC, Jude KM, Dong S, Ring AM, Votavova P, Pepper M, Kovar M, Garcia KC. Antibodies to interleukin-2 elicit selective T cell subset potentiation through distinct conformational mechanisms. Immunity. 2015;42:815–825. doi: 10.1016/j.immuni.2015.04.015. PubMed DOI PMC

Stepanek O, Prabhakar AS, Osswald C, King CG, Bulek A, Naeher D, Beaufils-Hugot M, Abanto ML, Galati V, Hausmann B, Lang R, Cole DK, Huseby ES, Sewell AK, Chakraborty AK, Palmer E. Coreceptor scanning by the T cell receptor provides a mechanism for T cell tolerance. Cell. 2014;159:333–345. doi: 10.1016/j.cell.2014.08.042. PubMed DOI PMC

Togashi Y, Shitara K, Nishikawa H. Regulatory T cells in cancer immunosuppression-implications for anticancer therapy. Nature Reviews. Clinical Oncology. 2019;16:356–371. doi: 10.1038/s41571-019-0175-7. PubMed DOI

Tomala J, Kovar M. IL-2/anti-IL-2 mAb immunocomplexes: a renascence of IL-2 in cancer immunotherapy? Oncoimmunology. 2016;5:e1102829. doi: 10.1080/2162402X.2015.1102829. PubMed DOI PMC

Wildin RS, Ramsdell F, Peake J, Faravelli F, Casanova JL, Buist N, Levy-Lahad E, Mazzella M, Goulet O, Perroni L, Bricarelli FD, Byrne G, McEuen M, Proll S, Appleby M, Brunkow ME. X-Linked neonatal diabetes mellitus, enteropathy and endocrinopathy syndrome is the human equivalent of mouse scurfy. Nature Genetics. 2001;27:18–20. doi: 10.1038/83707. PubMed DOI

Wong HS, Park K, Gola A, Baptista AP, Miller CH, Deep D, Lou M, Boyd LF, Rudensky AY, Savage PA, Altan-Bonnet G, Tsang JS, Germain RN. A local regulatory T cell feedback circuit maintains immune homeostasis by pruning self-activated T cells. Cell. 2021;184:3981–3997. doi: 10.1016/j.cell.2021.05.028. PubMed DOI PMC

Zheng GXY, Terry JM, Belgrader P, Ryvkin P, Bent ZW, Wilson R, Ziraldo SB, Wheeler TD, McDermott GP, Zhu J, Gregory MT, Shuga J, Montesclaros L, Underwood JG, Masquelier DA, Nishimura SY, Schnall-Levin M, Wyatt PW, Hindson CM, Bharadwaj R, Wong A, Ness KD, Beppu LW, Deeg HJ, McFarland C, Loeb KR, Valente WJ, Ericson NG, Stevens EA, Radich JP, Mikkelsen TS, Hindson BJ, Bielas JH. Massively parallel digital transcriptional profiling of single cells. Nature Communications. 2017;8:14049. doi: 10.1038/ncomms14049. PubMed DOI PMC

Temporal optimization of CD25-biased IL-2 agonists and immune checkpoint blockade leads to synergistic anticancer activity despite robust regulatory T cell expansion

ABIN1 is a negative regulator of effector functions in cytotoxic T cells

IL-2-driven CD8+ T cell phenotypes: implications for immunotherapy

Zobrazit více v PubMed

GEO
GSE183940