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IL-23 Promotes a Coordinated B Cell Germinal Center Program for Class-Switch Recombination to Igg2b in BXD2 Mice

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IL-23 Promotes a Coordinated B Cell Germinal Center Program for Class-Switch Recombination to IgG2b in BXD2 Mice This information is current as Huixian Hong, Min Gao, Qi Wu, PingAr Yang, Shanrun Liu, of October 2, 2021. Hao Li, Peter D. Burrows, Daniel Cua, Jake Y. Chen, Hui-Chen Hsu and John D. Mountz J Immunol published online 17 June 2020 http://www.jimmunol.org/content/early/2020/06/16/jimmun ol.2000280 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2020/06/16/jimmunol.200028 Material 0.DCSupplemental http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on October 2, 2021 *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 © 2020 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published June 17, 2020, doi:10.4049/jimmunol.2000280 The Journal of Immunology IL-23 Promotes a Coordinated B Cell Germinal Center Program for Class-Switch Recombination to IgG2b in BXD2 Mice Huixian Hong,* Min Gao,† Qi Wu,* PingAr Yang,* Shanrun Liu,* Hao Li,‡ Peter D. Burrows,x Daniel Cua,{,1 Jake Y. Chen,† Hui-Chen Hsu,* and John D. Mountz*,‖ IL-23 promotes autoimmune disease, including Th17 CD4 T cell development and autoantibody production. In this study, we show that a deficiency of the p19 component of IL-23 in the autoimmune BXD2 (BXD2-p192/2) mouse leads to a shift of the follicular T helper cell program from follicular T helper (Tfh)–IL-17 to Tfh–IFN-g. Although the germinal center (GC) size and the number of GC B cells remained the same, BXD2-p192/2 mice exhibited a lower class-switch recombination (CSR) in the GC B cells, leading to lower serum levels of IgG2b. Single-cell transcriptomics analysis of GC B cells revealed that whereas Ifngr1, Il21r, and Il4r genes exhibited a synchronized expression pattern with Cxcr5 and plasma cell program genes, Il17ra exhibited a synchronized Downloaded from expression pattern with Cxcr4 and GC program genes. Downregulation of Ighg2b in BXD2-p192/2 GC B cells was associated with decreased expression of CSR-related novel base excision repair genes that were otherwise predominantly expressed by Il17ra+ GC B cells in BXD2 mice. Together, these results suggest that although IL-23 is dispensable for GC formation, it is essential to promote a population of Tfh–IL-17 cells. IL-23 acts indirectly on Il17ra+ GC B cells to facilitate CSR-related base excision repair genes during the dark zone phase of GC B cell development. The Journal of Immunology, 2020, 205: 000–000. http://www.jimmunol.org/ ollicular T helper (Tfh) cells have been reported to express transcription factor and the cytokine IL-21 are key initial events well-characterized transcription factors and different cy- that enable the establishment of the Tfh cell program and upreg- F tokines that define this Th cell subset as well as molecules ulation of adhesion molecules, leading to effective, long-term, that promote T–B cell interactions (1, 2). Upregulation of the Bcl6 and productive interactions between T and B cells in the ger- minal center (GC) (3–6). The spectrum of CD4+CXCR5+PD- 1+ICOS+CD40L+ Tfh cells has increased over the past several *Division of Clinical Immunology and Rheumatology, Department of Medicine, years to include multiple subpopulations (5, 7, 8). Although University of Alabama at Birmingham, Birmingham, AL; †Informatics Institute, ‡ classical Tfh cells produce IL-21, sublineages and pathogenic Tfh the University of Alabama at Birmingham, Birmingham, AL; Beth Israel Deaconess by guest on October 2, 2021 Medical Center, Harvard Medical School, Boston, MA; xDepartment of Microbiol- cells, including IFN-g–producing Tfh–IFN-g, IL-4–producing ogy, University of Alabama at Birmingham, Birmingham, AL; {Discovery Research, ‖ Tfh–IL-4, and IL-17–producing Tfh–IL-17, have been identified Merck Research Laboratory, Boston, MA; and Department of Medicine, Birmingham VA Medical center, Birmingham, AL in both humans and mice (9–11). In BXD2 autoimmune mice, 1Current address: Janssen Pharmaceuticals, Spring House, PA IL-17 was shown to contribute to spontaneous GC formation and autoantibody production by enhancing and prolonging the inter- ORCIDs: 0000-0002-5592-9349 (M.G.); 0000-0002-7201-4212 (S.L.); 0000-0002- 2171-8826 (H.L.); 0000-0002-5752-5647 (P.D.B.); 0000-0001-8829-7504 (J.Y.C.); action between Tfh cells and B cells through the desensitization of 0000-0002-0317-5725 (H.-C.H.). chemokine receptor responses through regulator of G-protein Received for publication March 16, 2020. Accepted for publication May 14, 2020. signaling (RGS) 13 and RGS16 (5, 12–15). Such a response + + This work was supported by the National Institutes of Health (R01-AI-071110, R01 promotes the migration arrest of CXCR4 - or CXCR5 -respond- AI134023, and P30-AR-048311 to J.D.M. and R01-AI-083705 to H.-C.H.), U.S. ing cells and thereby is critical to enable prolonged interactions of Department of Veterans Affairs grants (I01BX004049 and 1I01BX000600 to J.D.M.), and the Lupus Research Alliance (a distinguished innovator award to J.D.M. and an GC B cells with other cells for GC B cell affinity maturation. alliance target identification award to H.-C.H.). Similarly, Tfh–IFN-g cells can promote the development of au- The single-cell RNA sequencing data presented in this article have been submitted to toreactive B cells outside the GC through the T-bet transcription the Gene Expression Omnibus database (https://www.ncbi.nlm.nih.gov/geo/query/ program (16). Despite these previous insights, it is not clear if acc.cgi?acc=GSE145922) under accession number GSE145922. these different Tfh cytokines operate concomitantly or sequen- Address correspondence and reprint requests to Dr. John D. Mountz, Department of Medicine, Division of Clinical Immunology and Rheumatology, University of tially in different niches or stages of GC B cell development to Alabama at Birmingham, Shelby Interdisciplinary Biomed Research Building, facilitate pathogenic autoantibody formation. It is also unknown if Room SHEL 307, 1825 University Boulevard, Birmingham, AL 35294-2182. the functions of one lineage can be replaced by the others. E-mail address: [email protected] It is also perplexing that activation-induced cytidine deaminase The online version of this article contains supplemental material. (AID) expression and activity primarily occurs in the highly Abbreviations used in this article: Ad, adenovirus; AID, activation-induced cytidine proliferative CXCR4+ dark zone (DZ) GC B cells, leading to deaminase; B6, C57BL/6; contig, contiguous sequence of overlapping DNA fragments; CSR, class-switch recombination; DZ, dark zone; GC, germinal center; KEGG, Kyoto extensive somatic hypermutation (SHM) (17, 18). In contrast, Tfh Encyclopedia of Genes and Genomes; LZ, light zone; NEIL, nucleotide excision repair- cells interact primarily with CXCR5+ GC B cells in the light zone like; NHEJ, nonhomologous end joining; PALS, periarteriolar lymphatic sheath; PB, plasmablast; PNA, peanut agglutinin; POLD, DNA polymerase d; qRT-PCR, quantitative (LZ) for fine-tuning of selection and plasma B cell development RT-PCR; RF, rheumatoid factor; RGS, regulator of G-protein signaling; scRNA-seq, (19–21). We previously have shown that the spontaneous autore- single-cell RNA sequencing; SHM, somatic hypermutation; Tfh, follicular T helper; active GCs in BXD2 mice lead to the development of autoanti- UAB, University of Alabama at Birmingham; WT, wild type. bodies through the upregulation of AID, which promotes SHM Copyright Ó 2020 by The American Association of Immunologists, Inc. 0022-1767/20/$37.50 and class-switch recombination (CSR) (22, 23). www.jimmunol.org/cgi/doi/10.4049/jimmunol.2000280 2 IL-23 PROMOTES A COORDINATED B CELL GC PROGRAM In addition to AID, both SHM and CSR require Bcl6-directed 660 anti-GL7 (clone GL7), FITC anti–PD-1 (clone J43), and Alexa Fluor programs of both B cell survival (24) and DNA damage and re- 647 anti–IL-17A (clone eBio17B7). Peanut agglutinin (PNA) was conju- pair genes (25–27). For CSR, nonhomologous end joining (NHEJ) gated with biotin (Vector Laboratories, Burlingame, CA) and detected by Alexa Fluor 488–conjugated streptavidin (Invitrogen). Dead cells were first requires transcription initiating upstream of the IgH constant excluded from analysis with allophycocyanin–eFluor 780 Organic Via- region undergoing CSR, followed by AID-mediated dsDNA bility Dye (eBioscience). breaks and then double-stranded break repair, resulting in a syn- For cytokine-producing T cell analysis, cells were stimulated for 5 h with apse and joining of the two switch regions (28). Known classical PMA (50 ng/ml; Sigma-Aldrich) and ionomycin (750 ng/ml; Sigma- Aldrich) in the presence of GolgiPlug (BD Biosciences). Cells were NHEJ players include Ku70 (XRCC6), Ku80 (XRCC5), XRCC4, stained for surface markers and then fixed and permeabilized with Cytofix/ DNA ligase IV, and DNA-PKcs (29), whereas the nonclassical (or Cytoperm solution (BD Biosciences) before intracellular staining (5). alternative) NHEJ pathway uses XRCC1 and DNA ligase III Data were acquired using standard flow cytometry (LSR II; BD Biosci- for DNA end joining (30, 31). Other DNA damage/repair gene ences) and analyzed with FlowJo_v10 software. FACS sorting was performed families, including histone variants H2a and H3a, which encode using a FACSAria cell sorter (BD Biosciences).
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    The Role of Histone Variants in the Epithelial-To-Mesenchymal Transition

    cells Review The Role of Histone Variants in the Epithelial-To-Mesenchymal Transition Imtiaz Nisar Lone 1, Burcu Sengez 1,2 , Ali Hamiche 3, Stefan Dimitrov 1,4 and Hani Alotaibi 1,2,* 1 Izmir Biomedicine and Genome Center, Izmir 35340, Turkey; [email protected] (I.N.L.); [email protected] (B.S.); [email protected] (S.D.) 2 Izmir International Biomedicine and Genome Institute, Dokuz Eylül University, Izmir 35340, Turkey 3 Institute of Genetics and Molecular and Cellular Biology (IGBMC), 1 rue Laurent Fries, 67400 Illkirch, France; [email protected] 4 Université Grenoble Alpes, CNRS UMR 5309, INSERM U1209, Institute for Advanced Biosciences (IAB), Site Santé-Allée des Alpes, 38700 La Tronche, France * Correspondence: [email protected]; Tel.: +90-232-299-4100 (ext. 5071) Received: 18 October 2020; Accepted: 14 November 2020; Published: 17 November 2020 Abstract: The epithelial-to-mesenchymal transition (EMT) is a physiological process activated during early embryogenesis, which continues to shape tissues and organs later on. It is also hijacked by tumor cells during metastasis. The regulation of EMT has been the focus of many research groups culminating in the last few years and resulting in an elaborate transcriptional network buildup. However, the implication of epigenetic factors in the control of EMT is still in its infancy. Recent discoveries pointed out that histone variants, which are key epigenetic players, appear to be involved in EMT control. This review summarizes the available data on histone variants’ function in EMT that would contribute to a better understanding of EMT itself and EMT-related diseases.