Unique patterns of CD8+ T-cell-mediated organ damage in the Act-mOVA/OT-I model of acute graft-versus-host disease

Cell Mol Life Sci. 2016 Oct;73(20):3935-47. doi: 10.1007/s00018-016-2237-7. Epub 2016 Apr 30.


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.

Keywords: CAG-OVA; Cytotoxic T cell; Experimental model; Homing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism*
  • Acute Disease
  • Animals
  • CD8-Positive T-Lymphocytes / immunology*
  • Cell Membrane / metabolism
  • Cell Proliferation
  • Cell Tracking
  • Chickens
  • Clone Cells
  • Disease Models, Animal
  • Flow Cytometry
  • Gene Expression Profiling
  • Graft vs Host Disease / immunology*
  • Graft vs Host Disease / pathology
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Organ Specificity*
  • Ovalbumin / metabolism*
  • Reproducibility of Results
  • T-Lymphocytes, Cytotoxic / immunology


  • Actins
  • Ovalbumin