Eph/ephrin-B-mediated cell-to-cell interactions govern MTS20+ thymic epithelial cell development

Research Projects
Organizational Units
Journal Issue
Thymus development is a complex process in which cell-to-cell interactions between thymocytes and thymic epithelial cells (TECs) are essential to allow a proper maturation of both thymic cell components. Although signals that control thymocyte development are well known, mechanisms governing TEC maturation are poorly understood, especially those that regulate the maturation of immature TEC populations during early fetal thymus development. In this study, we show that EphB2-deficient, EphB2LacZ and EphB3-deficient fetal thymuses present a lower number of cells and delayed maturation of DN cell subsets compared to WT values. Moreover, deficits in the production of chemokines, known to be involved in the lymphoid seeding into the thymus, contribute in decreased proportions of intrathymic T cell progenitors (PIRA/B+) in the mutant thymuses from early stages of development. These features correlate with increased proportions of MTS20+ cells but fewer MTS20− cells from E13.5 onward in the deficient thymuses, suggesting a delayed development of the first epithelial cells. In addition, in vitro the lack of thymocytes or the blockade of Eph/ephrin-B-mediated cell-to-cell nteractions between either thymocytes–TECs or TECs–TECs in E13.5 fetal thymic lobes coursed with increased proportions of MTS20+ TECs. This confirms, for the first time, that the presence of CD45+ cells, corresponding at these stages to DN1 and DN2 cells, and Eph/ephrin-B-mediated heterotypic or homotypic cell interactions between thymocytes and TECs, or between TECs and themselves, contribute to the early maturation of MTS20+ TECs.
Aasheim HC, Delabie J, Finne EF (2005) Ephrin-A1 binding toCD4+ T lymphocytes stimulates migration and induces tyrosinephosphorylation of PYK2. Blood 105:2869–2876. doi:10.1182/blood-2004-08-2981 Alfaro D et al (2007) EphrinB1-EphB signaling regulates thymocyteepithelium interactions involved in functional T cell development.Eur J Immunol 37:2596–2605.doi:10.1002/eji.200737097 Alfaro D, Munoz JJ, Garcia-Ceca J, Cejalvo T, Jimenez E, Zapata A (2008) Alterations in the thymocyte phenotype of EphB-deficient Histochem Cell Biol (2016) 146:167–182 181 1 3 mice largely affect the double negative cell compartment. Immunology 125:131–143. doi:10.1111/j.1365-2567.2008.02828.x Alfaro D, Munoz JJ, Garcia-Ceca J, Cejalvo T, Jimenez E, Zapata AG (2011) The Eph/ephrinB signal balance determines the pattern of T-cell maturation in the thymus. Immunol Cell Biol 89:844–852.doi:10.1038/icb.2010.172 Alfaro D et al (2015) EphB2 and EphB3 play an important role in the lymphoid seeding of murine adult thymus. J Leukoc Biol 98:883–896. doi:10.1189/jlb.1HI1114-568R Anderson G, Jenkinson EJ (2001) Lymphostromal interactions in thymic development and function. Nat Rev Immunol 1:31–40.doi:10.1038/35095500 Anderson G, Jenkinson EJ, Rodewald HR (2009) A roadmap for thymic epithelial cell development. Eur J Immunol 39:1694– 1699. doi:10.1002/eji.200939379 Bennett AR, Farley A, Blair NF, Gordon J, Sharp L, Blackburn CC (2002) Identification and characterization of thymic epithelial progenitor cells. Immunity 16:803–814 Blackburn CC, Manley NR (2004) Developing a new paradigm for thymus organogenesis. Nat Rev Immunol 4:278–289. doi:10.1038/nri1331 Blackburn CC et al (1996) The nu gene acts cell-autonomously and is required for differentiation of thymic epithelial progenitors. Proc Natl Acad Sci USA 93:5742–5746 Boehm T, Scheu S, Pfeffer K, Bleul CC (2003) Thymic medullary epithelial cell differentiation, thymocyte emigration, and the control of autoimmunity require lympho-epithelial cross talk via LTbetaR. J Exp Med 198:757–769. doi:10.1084/jem.20030794 Calderon L, Boehm T (2011) Three chemokine receptors cooperatively regulate homing of hematopoietic progenitors to the embryonic mouse thymus. Proc Natl Acad Sci USA 108:7517–7522. doi:10.1073/pnas.1016428108 Colucci F, Soudais C, Rosmaraki E, Vanes L, Tybulewicz VL, Di Santo JP (1999) Dissecting NK cell development using a novel alymphoid mouse model: investigating the role of the c-abl proto-oncogene in murine NK cell differentiation. J Immunol 162:2761–2765 Davy A, Soriano P (2005) Ephrin signaling in vivo: look both ways.Dev Dyn 232:1–10. doi:10.1002/dvdy.20200 Depreter MG et al (2008) Identification of Plet-1 as a specific marker of early thymic epithelial progenitor cells. Proc Natl Acad Sci USA 105:961–966.doi:10.1073/pnas.0711170105 Garcia-Ceca J et al (2009) On the role of Eph signalling in thymus histogenesis; EphB2/B3 and the organizing of the thymic epithelial network. Int J Dev Biol 53:971–982. doi:10.1387/ijdb.082702jg Garcia-Ceca J, Alfaro D, Montero-Herradon S, Zapata AG (2013) Eph/ephrinB signalling is involved in the survival of thymic epithelial cells. Immunol Cell Biol 91:130–138. doi:10.1038/icb.2012.59 Garcia-Ceca J, Alfaro D, Montero-Herradon S, Tobajas E, Munoz JJ,Zapata AG (2015) Eph/ephrins-mediated thymocyte-thymic epithelial cell interactions control numerous processes of thymus biology. Front Immunol 6:333. doi:10.3389/fimmu.2015.00333 Gill J, Malin M, Hollander GA, Boyd R (2002) Generation of a complete thymic microenvironment by MTS24(+) thymic epithelial cells. Nat Immunol 3:635–642. doi:10.1038/ni812 Godfrey DI, Izon DJ, Tucek CL, Wilson TJ, Boyd RL (1990) The phenotypic heterogeneity of mouse thymic stromal cells. Immunology70:66–74 Henkemeyer M, Orioli D, Henderson JT, Saxton TM, Roder J, Pawson T, Klein R (1996) Nuk controls pathfinding of commissural axons in the mammalian central nervous system. Cell 86:35–46 Hjorthaug HS, Aasheim HC (2007) Ephrin-A1 stimulates migration of CD8+ CCR7+ T lymphocytes. Eur J Immunol 37:2326–2336. doi:10.1002/eji.200737111 Hollander GA et al (1995) Developmental control point in induction of thymic cortex regulated by a subpopulation of prothymocytes.Nature 373:350–353. doi:10.1038/373350a0 Itoi M, Kawamoto H, Katsura Y, Amagai T (2001) Two distinct steps of immigration of hematopoietic progenitors into the early thymus anlage. Int Immunol 13:1203–1211 Jenkinson WE, Rossi SW, Parnell SM, Agace WW, Takahama Y, Jenkinson EJ, Anderson G (2007) Chemokine receptor expression defines heterogeneity in the earliest thymic migrants. Eur J Immunol 37:2090–2096. doi:10.1002/eji.200737212 Jotereau F, Heuze F, Salomon-Vie V, Gascan H (1987) Cell kinetics in the fetal mouse thymus: precursor cell input, proliferation, and emigration. J Immunol 138:1026–1030 Kitamura T et al (2008) Enhancement of lymphocyte migration and cytokine production by ephrinB1 system in rheumatoid arthritis. Am J Physiol Cell Physiol 294:C189–C196. doi:10.1152/ajpcell.00314.2007 Klug DB, Carter C, Crouch E, Roop D, Conti CJ, Richie ER (1998) Interdependence of cortical thymic epithelial cell differentiation and T-lineage commitment. Proc Natl Acad Sci USA 95:11822–11827 Klug DB, Carter C, Gimenez-Conti IB, Richie ER (2002) Cuttingedge: thymocyte-independent and thymocyte-dependent phases of epithelial patterning in the fetal thymus. J Immunol 169:2842–2845 Knoll B, Drescher U (2002) Ephrin-As as receptors in topographic projections. Trends Neurosci 25:145–149 Laurent J, Bosco N, Marche PN, Ceredig R (2004) New insights into the proliferation and differentiation of early mouse thymocytes.Int Immunol 16:1069–1080. doi:10.1093/intimm/dxh108 Liu C et al (2005) The role of CCL21 in recruitment of T-precursor cells to fetal thymi. Blood 105:31–39. doi:10.1182/blood-2004-04-1369 Liu C et al (2006) Coordination between CCR7- and CCR9-mediated chemokine signals in prevascular fetal thymus colonization.Blood 108:2531–2539. doi:10.1182/blood-2006-05-024190 Munoz JJ, Cejalvo T, Alonso-Colmenar LM, Alfaro D, Garcia-Ceca J, Zapata A (2011) Eph/ephrin-mediated interactions in the thymus.NeuroImmunoModulation 18:271–280. doi:10.1159/000329490 Pasquale EB (2008) Eph-ephrin bidirectional signaling in physiology and disease. Cell 133:38–52.doi:10.1016/j.cell.2008.03.011 Perez White BE, Getsios S (2014) Eph receptor and ephrin function in breast, gut, and skin epithelia. Cell Adhes Migr 8:327–338. doi:10.4161/19336918.2014.970012 Romano R, Palamaro L, Fusco A, Giardino G, Gallo V, Del Vecchio L, Pignata C (2013) FOXN1: a master regulator gene of thymic epithelial development program. Front Immunol 4:187.doi:10.3389/fimmu.2013.00187 Rossi SW, Jenkinson WE, Anderson G, Jenkinson EJ (2006) Clonal analysis reveals a common progenitor for thymic cortical and medullary epithelium. Nature 441:988–991. doi:10.1038/nature04813 Rossi SW et al (2007a) Redefining epithelial progenitor potential in the developing thymus. Eur J Immunol 37:2411–2418.doi:10.1002/eji.200737275 Rossi SW et al (2007b) RANK signals from CD4(+)3(−) inducer cells regulate development of Aire-expressing epithelial cells in the thymic medulla. J Exp Med 204:1267–1272. doi:10.1084/jem.20062497 Shakib S, Desanti GE, Jenkinson WE, Parnell SM, Jenkinson EJ,Anderson G (2009) Checkpoints in the development of thymic cortical epithelial cells. J Immunol 182:130–137 Sharfe N, Freywald A, Toro A, Dadi H, Roifman C (2002) Ephrin stimulationmodulates T cell chemotaxis. Eur J Immunol 32:3745–3755. doi:10.1002/1521-4141(200212)32:12<3745::AIDIMMU3745>3.0.CO;2-M Stimamiglio MA et al (2010) EphB2-mediated interactions are essential for proper migration of T cell progenitors during fetal thymus colonization. J Leukoc Biol 88:483–494. doi:10.1189/jlb.0210079 Trinidad EM, Ballesteros M, Zuloaga J, Zapata A, Alonso-Colmenar LM (2009) An impaired transendothelial migration potential of chronic lymphocytic leukemia (CLL) cells can be linked to ephrin-A4 expression. Blood 114:5081–5090. doi:10.1182/blood-2009-03-210617 van Ewijk W, Shores EW, Singer A (1994) Crosstalk in the mouse thymus. Immunol Today 15:214–217.doi:10.1016/0167-5699(94)90246-1 van Ewijk W, Hollander G, Terhorst C, Wang B (2000) Stepwise development of thymic microenvironments in vivo is regulated by thymocyte subsets. Development 127:1583–1591 Wang JH, Nichogiannopoulou A, Wu L, Sun L, Sharpe AH, Bigby M, Georgopoulos K (1996) Selective defects in the development of the fetal and adult lymphoid system in mice with an Ikaros null mutation. Immunity 5:537–549 Wilkinson B, Owen JJ, Jenkinson EJ (1999) Factors regulating stem cell recruitment to the fetal thymus. J Immunol 162:3873–3881 Zamora DO, Babra B, Pan Y, Planck SR, Rosenbaum JT (2006) Human leukocytes express ephrinB2 which activates microvascular endothelial cells. Cell Immunol 242:99–109. doi:10.1016/j.cellimm.2006.10.001 Zhu M, Fu YX (2008) Coordinating development of medullary thymic epithelial cells. Immunity 29:386–88.doi:10.1016/j. immuni.2008.09.001 Zlotoff DA, Bhandoola A (2011) Hematopoietic progenitor migration to the adult thymus. Ann N Y Acad Sci 1217:122–138.doi:10.1111/j.1749-6632.2010.05881.x