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A Hypermorphic Nfkbid Allele Contributes to Impaired Thymic Deletion of Autoreactive Diabetogenic CD8+ T Cells in NOD Mice

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A Hypermorphic Nfkbid Allele Contributes to Impaired Thymic Deletion of Autoreactive Diabetogenic CD8+ T Cells in NOD Mice A Hypermorphic Nfkbid Allele Contributes to Impaired Thymic Deletion of Autoreactive Diabetogenic CD8+ T Cells in NOD Mice This information is current as Maximiliano Presa, Jeremy J. Racine, Jennifer R. Dwyer, of October 2, 2021. Deanna J. Lamont, Jeremy J. Ratiu, Vishal Kumar Sarsani, Yi-Guang Chen, Aron Geurts, Ingo Schmitz, Timothy Stearns, Jennifer Allocco, Harold D. Chapman and David V. Serreze J Immunol published online 20 August 2018 http://www.jimmunol.org/content/early/2018/08/17/jimmun Downloaded from ol.1800465 Supplementary http://www.jimmunol.org/content/suppl/2018/08/19/jimmunol.180046 http://www.jimmunol.org/ Material 5.DCSupplemental Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on October 2, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2018 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published August 20, 2018, doi:10.4049/jimmunol.1800465 The Journal of Immunology A Hypermorphic Nfkbid Allele Contributes to Impaired Thymic Deletion of Autoreactive Diabetogenic CD8+ T Cells in NOD Mice Maximiliano Presa,* Jeremy J. Racine,* Jennifer R. Dwyer,* Deanna J. Lamont,* Jeremy J. Ratiu,* Vishal Kumar Sarsani,* Yi-Guang Chen,† Aron Geurts,† Ingo Schmitz,‡,x Timothy Stearns,* Jennifer Allocco,* Harold D. Chapman,* and David V. Serreze* In both NOD mice and humans, the development of type 1 diabetes (T1D) is dependent in part on autoreactive CD8+ T cells recognizing pancreatic b cell peptides presented by often quite common MHC class I variants. Studies in NOD mice previously d b revealed that the common H2-K and/or H2-D class I molecules expressed by this strain aberrantly lose the ability to mediate the Downloaded from thymic deletion of pathogenic CD8+ T cell responses through interactions with T1D susceptibility genes outside the MHC. A gene (s) mapping to proximal chromosome 7 was previously shown to be an important contributor to the failure of the common class I molecules expressed by NOD mice to mediate the normal thymic negative selection of diabetogenic CD8+ T cells. Using an inducible model of thymic negative selection and mRNA transcript analyses, we initially identified an elevated Nfkbid expression variant as a likely NOD-proximal chromosome 7 region gene contributing to impaired thymic deletion of diabetogenic CD8+ T cells. CRISPR/Cas9–mediated genetic attenuation of Nfkbid expression in NOD mice resulted in improved negative selection of http://www.jimmunol.org/ autoreactive diabetogenic AI4 and NY8.3 CD8+ T cells. These results indicated that allelic variants of Nfkbid contribute to the efficiency of intrathymic deletion of diabetogenic CD8+ T cells. However, although enhancing thymic deletion of pathogenic CD8+ T cells, ablating Nfkbid expression surprisingly accelerated T1D onset that was associated with numeric decreases in both regulatory T and B lymphocytes in NOD mice. The Journal of Immunology, 2018, 201: 000–000. n both the NOD mouse model and humans, type 1 diabetes Some Idd genes appear to contribute to defects in central tolerance (T1D) results from the autoimmune destruction of insulin- mechanisms that normally represent a first checkpoint, engender- producing pancreatic b cells mediated by the combined ac- ing the thymic deletion of autoreactive CD4+ and CD8+ Tcells by guest on October 2, 2021 I + + tivity of CD4 and CD8 T cells as well as B lymphocytes (1). T1D during early stages of their development (6–8). However, even in is highly polygenic in nature, as more than 50 loci have been as- healthy individuals, some autoreactive T cells escape central toler- sociated with disease susceptibility or resistance in both humans ance mechanisms (9). Such autoreactive effectors are normally and NOD mice (2, 3). Although its complete pathogenic etiology prevented from mediating pathogenic effects by a second checkpoint remains unsolved, there is a wide acceptance that disease develops provided by multiple mechanisms of peripheral tolerance involving as a consequence of interactions between allelically variable genes regulatory lymphocyte populations (regulatory T lymphocytes contributing to T1D susceptibility or resistance (Idd), respectively, [Tregs]/regulatory B lymphocytes [Bregs]), with significant evi- resulting in breakdowns and maintenance of mechanisms control- dence for some of these processes also being disrupted by various ling induction of immunological tolerance to self-proteins (4, 5). Idd genes (5). *The Jackson Laboratory, Bar Harbor, ME 04609; †Medical College of Wisconsin, experiments, interpreted data, and contributed to writing the manuscript. J.J. Ratiu, Milwaukee, WI 53226; ‡Systems-Oriented Immunology and Inflammation Research D.J.L., J.A., and H.D.C. conducted experimentation. V.K.S. and T.S. contributed to Group, Helmholtz Centre for Infection Research, 38124 Braunschweig, Germany; statistical analyses. Y.-G.C., A.G., and I.S. contributed to experimental design. D.V.S. and xInstitute of Molecular and Clinical Immunology, Otto-von-Guericke University, contributed to study conception and supervised experimental effort and writing of the 39120 Magdeburg, Germany manuscript. ORCIDs: 0000-0002-0160-6528 (M.P.); 0000-0001-5514-3074 (J.J. Racine); 0000- The microarray data presented in this article have been submitted to the Gene 0002-5893-6026 (J.R.D.); 0000-0002-9272-3438 (V.K.S.); 0000-0002-5360- Expression Omnibus under accession number GSE115754. 0419 (I.S.); 0000-0001-7614-5925 (D.V.S.). Address correspondence and reprint requests to Dr. David V. Serreze, The Jackson Received for publication March 27, 2018. Accepted for publication July 23, 2018. Laboratory, 600 Main Street, Bar Harbor, ME 04609. E-mail address: dave. [email protected] M.P. was supported by JDRF Fellowship 3-PDF-2014-219-A-N. For parts of this work, J.J. Racine was supported by either National Institutes of Health (NIH) Fel- The online version of this article contains supplemental material. lowship 1F32DK111078 or JDRF Fellowship 3-PDF-2017-372-A-N. J.J. Racine was Abbreviations used in this article: B6, C57BL/6J; Breg, regulatory B lymphocyte; also supported by a grant from the Diabetes Research Connection (DRC 006887 JR). Chr, chromosome; DP, CD4+CD8+ double positive; FDR, false discovery rate; Idd, D.V.S. is supported by NIH Grants DK-46266, DK-95735, and OD-020351-5022 as allelically variable gene contributing to T1D susceptibility or resistance; LCMV, well as by JDRF Grant 2018-568. This work was also partly supported by Cancer lymphocytic choriomeningitis virus; MZ, marginal zone; NOD, NOD/ShiLtDvs; Center Support Grant CA34196. I.S. was supported by grants from the Deutsche 2 2 , . NOD-Nfkbid / , NOD/ShiLtDvs-Nfkbid em3Dvs /Dvs; qPCR, quantitative PCR; Forschungsgemeinschaft (SCHM1586/6-1 and Project A23 of SFB854). A generous sgRNA, single guide RNA; SNP, single nucleotide polymorphism; T1D, type 1 donation from Carl Stiewe and wife Maike Rohde toward type 1 diabetes research diabetes; Tet-AI4, H2-Db/MimA2 tetramer capable of binding the AI4 TCR; Treg, at The Jackson Laboratory contributed to this work. regulatory T lymphocyte. M.P. designed and conducted experimentation, interpreted data, and wrote the man- uscript. J.J. Racine designed and conducted experimentation and contributed to Copyright Ó 2018 by The American Association of Immunologists, Inc. 0022-1767/18/$35.00 data interpretation and writing of the manuscript. J.R.D. designed and conducted www.jimmunol.org/cgi/doi/10.4049/jimmunol.1800465 2 Nfkbid CONTROLS THYMIC DELETION OF DIABETOGENIC CD8 T CELLS Immature CD4+CD8+ double positive (DP) thymocytes transgenic mouse models, we show that a higher Nfkbid expression expressing TCR molecules allowing for high avidity interactions variant in NOD compared with B6 mice contributes to what is likely with APC displaying self-peptide–MHC complexes are normally diminished negative selection in the former strain of both the AI4 deleted by apoptosis or diverted to a CD4+Foxp3+ Treg lineage and NY8.3 MHC class I–restricted autoreactive diabetogenic CD8+ (9). Idd gene-elicited deficiencies in this process contribute to the T cell clonotypes. Although genetic attenuation of Nfkbid expres- thymic survival and peripheral seeding of both autoreactive CD4+ sion to the lower levels observed in B6 background mice results in and CD8+ T cells that overwhelm the suppressive capacity of self- improved thymic deletion of pathogenic CD8+ T cells, total ablation antigen–specific Tregs (4, 5). MHC class II–restricted autoreactive of this gene also numerically diminishes levels of peripheral Tregs CD4+ T cells clearly play a critical role in T1D development. and Bregs associated with a more rapid onset of T1D in the mutant However, they cannot do so by directly engaging insulin- stock than in standard NOD mice. producing pancreatic b cells that only express MHC class I molecules. Hence, through an ability to directly engage and de- Materials and Methods stroy pancreatic b cells, MHC class I–restricted autoreactive Mouse strains CD8+ T cells are likely the ultimate mediators of T1D develop- NOD/ShiLtDvs (hereafter NOD), B6, and NOD.Cg-Prkdcscid Emv30b/Dvs ment in both humans and NOD mice (10, 11). Indeed, epidemi- (NOD.scid) mice are maintained in a specific pathogen-free research colony ological studies have shown that in addition to class II effects, at The Jackson Laboratory. NOD mice carrying a transgenic Va8.3+Vb2+ particular HLA class I variants provide an independent T1D risk TCR derived from the AI4 diabetogenic CD8+ T cell clone (NOD-AI4) have factor in humans (12, 13).
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