title: Unique patterns of CD8+ T-cell-mediated organ damage in the Act-mOVA/OT-I model of acute graft-versus-host disease.
abstract: T-cell receptor (TCR)-transgenic models of acute graft-versus-host disease (aGvHD) offer a straightforward and highly controlled approach to study the mechanisms and consequences of T-cell activation following allogeneic hematopoietic stem cell transplantation (aHSCT). Here, we report that aHSCT involving OT-I mice as donors, carrying an ovalbumin-specific CD8+ TCR, and Act-mOVA mice as recipients, expressing membrane-bound ovalbumin driven by the β-actin promoter, induces lethal aGvHD in a CD8+ T-cell-dependent, highly reproducible manner, within 4-7 days. Tracking of UBC-GFP/OT-I graft CD8+ T cells disclosed heavy infiltration of the gastrointestinal tract, liver, and lungs at the onset of the disease, and histology confirmed hallmark features of gastrointestinal aGVHD, hepatic aGvHD, and aGvHD-associated lymphocytic bronchitis in infiltrated organs. However, T-cell infiltration was virtually absent in the skin, a key target organ of human aGvHD, and histology confirmed the absence of cutaneous aGVHD, as well. We show that the model allows studying CD8+ T-cell responses in situ, as selective recovery of graft CD45.1/OT-I CD8+ T cells from target organs is simple and feasible by automated tissue dissociation and subsequent cell sorting. Assessment of interferon-gamma production by flow cytometry, granzyme-B release by ELISA, TREC assay, and whole-genome gene expression profiling confirmed that isolated graft CD8+ T cells remained intact, underwent clonal expansion, and exerted effector functions in all affected tissues. Taken together, these data demonstrate that the OT-I/Act-mOVA model is suitable to study the CD8+ T-cell-mediated effector mechanisms in a disease closely resembling fatal human gastrointestinal and hepatic aGVHD that may develop after aHSCT using HLA-matched unrelated donors.
authors.affiliation: Hungarian Academy of Sciences-Semmelweis University, "Lendület" Experimental and Translational Immunomics Research Group, 1089, Budapest, Hungary.|Department of Genetics, Cell and Immunobiology, Semmelweis University, 1089, Budapest, Hungary.
authors.forename: Barbara
authors.initials: B
authors.lastname: Érsek
authors.affiliation: Hungarian Academy of Sciences-Semmelweis University, "Lendület" Experimental and Translational Immunomics Research Group, 1089, Budapest, Hungary.|Department of Genetics, Cell and Immunobiology, Semmelweis University, 1089, Budapest, Hungary.
authors.forename: Nikolett
authors.initials: N
authors.lastname: Lupsa
authors.affiliation: 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, 1085, Budapest, Hungary.
authors.forename: Péter
authors.initials: P
authors.lastname: Pócza
authors.affiliation: Hungarian Academy of Sciences-Semmelweis University, "Lendület" Experimental and Translational Immunomics Research Group, 1089, Budapest, Hungary.|Department of Genetics, Cell and Immunobiology, Semmelweis University, 1089, Budapest, Hungary.
authors.forename: Anett
authors.initials: A
authors.lastname: Tóth
authors.affiliation: Hungarian Academy of Sciences-Semmelweis University, "Lendület" Experimental and Translational Immunomics Research Group, 1089, Budapest, Hungary.|Department of Genetics, Cell and Immunobiology, Semmelweis University, 1089, Budapest, Hungary.
authors.forename: Andor
authors.initials: A
authors.lastname: Horváth
authors.affiliation: Department of Genetics, Cell and Immunobiology, Semmelweis University, 1089, Budapest, Hungary.|Csertex Research Laboratory, 1037, Budapest, Hungary.
authors.forename: Viktor
authors.initials: V
authors.lastname: Molnár
authors.affiliation: Hungarian Academy of Sciences-Semmelweis University, "Lendület" Experimental and Translational Immunomics Research Group, 1089, Budapest, Hungary.|Department of Genetics, Cell and Immunobiology, Semmelweis University, 1089, Budapest, Hungary.
authors.forename: Bence
authors.initials: B
authors.lastname: Bagita
authors.affiliation: Hungarian Academy of Sciences-Semmelweis University, "Lendület" Experimental and Translational Immunomics Research Group, 1089, Budapest, Hungary.|Department of Genetics, Cell and Immunobiology, Semmelweis University, 1089, Budapest, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Bencsik
authors.affiliation: "Frédéric Joliot-Curie" Institute for Radiobiology and Radiohygiene, 1221, Budapest, Hungary.
authors.forename: Hargita
authors.initials: H
authors.lastname: Hegyesi
authors.affiliation: 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, 1085, Budapest, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Matolcsy
authors.affiliation: Department of Genetics, Cell and Immunobiology, Semmelweis University, 1089, Budapest, Hungary.
authors.forename: Edit I
authors.initials: EI
authors.lastname: Buzás
authors.affiliation: Hungarian Academy of Sciences-Semmelweis University, "Lendület" Experimental and Translational Immunomics Research Group, 1089, Budapest, Hungary. pos.zoltan@med.semmelweis-univ.hu.|Department of Genetics, Cell and Immunobiology, Semmelweis University, 1089, Budapest, Hungary. pos.zoltan@med.semmelweis-univ.hu.
authors.forename: Zoltán
authors.identifier: 0000-0002-2574-7616
authors.initials: Z
authors.lastname: Pós
mesh_terms: D000199:Actins / Q000378:metabolism*; D000208:Acute Disease; D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D058948:Cell Tracking; D002645:Chickens; D002999:Clone Cells; D004195:Disease Models, Animal; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006086:Graft vs Host Disease / Q000276:immunology* / Q000473:pathology; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009928:Organ Specificity; D010047:Ovalbumin / Q000378:metabolism*; D015203:Reproducibility of Results; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D000199:Actins; D010047:Ovalbumin
keywords: CAG-OVA; Cytotoxic T cell; Experimental model; Homing