Chronic graft-versus-host-disease in CD34 þ -humanized NSG mice is associated with human susceptibility HLA haplotypes for autoimmune disease Katja Sonntag a, 1 , Franziska Eckert a, b, 1 , Christian Welker a , Hartmut Müller c , Friederike Müller a , Daniel Zips b , Bence Sipos c , Reinhild Klein d , Gregor Blank a , Tobias Feuchtinger e , Michael Schumm a , Rupert Handgretinger a , Karin Schilbach a, * a Department of General Pediatrics, Oncology/Hematology, Eberhard Karls University Tübingen, Hoppe-Seyler-Str.1, 72076 Tübingen, Germany b Department of Radiation Oncology, Eberhard Karls University Tübingen, Hoppe-Seyler-Str. 3, 72076 Tübingen, Germany c Institute of Pathology, Eberhard Karls University Tübingen, Liebermeisterstraße 8, 72076 Tübingen, Germany d Laboratory for Immunopathology, Eberhard Karls University Tübingen, Otfried-Müller-Straße 10, 72076 Tübingen, Germany e Pediatric Hematology, Oncology and Stem Cell Transplantation Dr. von Hauner'sches Kinderspital, Ludwig-Maximilian-University Munich, Lindwurmstraße 4, 80337 München, Germany article info Article history: Received 2 March 2015 Received in revised form 27 May 2015 Accepted 7 June 2015 Available online xxx Keywords: Chronic graft-versus-host disease Autoimmunity cGVHD model cGVHD pathology Extended HLA haplotypes T-cell-receptor repertoire Humanized mouse abstract Chronic graft-versus-host disease (cGVHD) is a significant hurdle to long-term hematopoietic stem-cell transplantation success. Insights into the pathogenesis and mechanistical investigations of novel ther- apeutic strategies are limited as appropriate animal models are missing. The immunodeficient NSG mouse e when humanized with human bone marrow, fetal liver and thymus (BLT NSG) e is prone for cGVHD, yet mainly affects the skin. In contrast, the NSG mouse humanized exclusively with CD34 þ - selected, CD3 þ -depleted stem cells (CD34 þ NSG) has neither been described for acute nor chronic GVHD so far. This is the first report about the development of systemic autoimmune cGVHD 24 weeks post stem cell receipt involving lung, liver, skin, gingiva and intestine in two NSG cohorts humanized with CD34 þ grafts from different donors. Affected mice presented with sclerodermatous skin, fibrotic lung, severe hepatitis, and massive dental malformation/loss. CD4 þ -dominated, T H 2-biased, bulky T-cell in- filtrates featured highly skewed T cell receptor (TCR) repertoires, clonal expansions, and autoreactive TCRs. In affected tissues profibrotic IL-13 and -4 dominated over T H 1 cytokines IFN-g and TNF-a. Thus, the time point of manifestation and the phenotype match human systemic pleiotropic sclerodermatous GVHD. The CD34 þ NSG-model's intrinsic deficiency of thymus, thymus-derived regulatory T cells (nTreg) and B cells emphasizes the role of the genetic polymorphism and the cytokines in the pathogenesis of cGVHD. Importantly, the only factor discriminating diseased versus non-diseased CD34 þ NSG cohorts were two risk HLA haplotypes that in human mediate susceptibility for autoimmune disease (psoriasis). Thus, the CD34 þ NSG model may serve as a platform for addressing issues related to the pathophysiology and treatment of human autoimmunity and chronic GVHD. © 2015 Elsevier Ltd. All rights reserved. 1. Introduction Chronic graft-versus-host disease (cGVHD) is a significant hur- dle to long-term hematopoietic stem cell transplantation success. Effective treatments for cGVHD have been difficult to develop, since exact causes for the disease are enigmatic. This is in part due to the inability of animal models to recapitulate the multiorgan pathol- ogies observed in clinical cGVHD [1]. Acute GVHD (aGVHD) results from an immune-mediated attack on recipient tissue by donor T cells contained in or developed from the graft [2,3] that recognize and bind their cognate ligands on recipient antigen-presenting cells (APC) in combination with essential second signals on the APC's surface. The resulting * Corresponding author. University Children's Hospital, Clinical and Experimental Immunology, Hoppe-Seyler-Str. 1, 72076 Tübingen, Germany. E-mail address: karin.schilbach@med.uni-tuebingen.de (K. Schilbach). 1 Contributed equally to the work. Contents lists available at ScienceDirect Journal of Autoimmunity journal homepage: www.elsevier.com/locate/jautimm http://dx.doi.org/10.1016/j.jaut.2015.06.006 0896-8411/© 2015 Elsevier Ltd. All rights reserved. Journal of Autoimmunity xxx (2015) 1e12 Please cite this article in press as: K. Sonntag, et al., Chronic graft-versus-host-disease in CD34 þ -humanized NSG mice is associated with human susceptibility HLA haplotypes for autoimmune disease, Journal of Autoimmunity (2015), http://dx.doi.org/10.1016/j.jaut.2015.06.006