Open Access Research article

A diagnostic window for the treatment of acute graft-versus-host disease prior to visible clinical symptoms in a murine model

Carina A Bäuerlein123, Simone S Riedel123, Jeanette Baker1234, Christian Brede123, Ana-Laura Jordán Garrote123, Martin Chopra12, Miriam Ritz12, Georg F Beilhack5, Stephan Schulz6, Robert Zeiser7, Paul G Schlegel38, Hermann Einsele13, Robert S Negrin4 and Andreas Beilhack123*

Author Affiliations

1 Department of Medicine II, Würzburg University Clinics, Zinklesweg 10, Würzburg, D-97078, Germany

2 Interdisciplinary Center for Clinical Research (IZKF), Würzburg, Germany

3 Graduate School of Life Sciences, Würzburg, Germany

4 Division of Blood and Marrow Transplantation, Department of Medicine, Stanford University, Stanford, CA, USA

5 Department of Medicine III, Vienna Medical University, Vienna, Austria

6 Institute of Pathology Charité, Berlin, Germany

7 Department of Hematology and Oncology, Freiburg University Medical Center, Freiburg, Germany

8 Department of Pediatrics, Würzburg University Hospital, Würzburg, Germany

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BMC Medicine 2013, 11:134  doi:10.1186/1741-7015-11-134

Published: 21 May 2013

Additional files

Additional file 1:

Initial donor T cell proliferation and migration after allogeneic hematopoietic cell transplantation (allo-HCT) follows the same spatiotemporal pattern but differs in bioluminescence imaging (BLI) signal increase in the major histocompatibility complex (MHC) versus the minor histocompatibility antigen (miHAg) mismatched mouse model. (A)Luc+ donor T cells proliferate in secondary lymphoid organs (SLOs) until day +3 in both allogeneic models. They show the same spatiotemporal shift from acute graft-versus-host disease (aGVHD) initiation to effector phase but in the miHAg mismatch model (middle panel) signals increase less intensely compared to the MHC major mismatch model (upper panel) during the first 7 days followed by a strong increase in whole body signal intensity from day +8 on. In syngeneic transplanted albino B6 mice (lower panel) donor T cells initially also home to SLOs but as well to the thymus and bone marrow (BM) and not to the skin. One representative mouse per group (n = 6) is shown. (B) Quantification of dynamic total body and single organ BLI signal changes. Photon emissions per second per whole animal and average photon radiation for representative target organs are displayed. Total body BLI signals increase dramatically in mice with aGVHD in both allogeneic models, MHC major (●) and miHAg (■) mismatched groups, but only moderately in syngeneic (▼) transplanted mice. Overall gastrointestinal tract (GIT) and skin signal intensities of the MHC major mismatched mice increase much stronger than in the miHAg mismatched model before those mice die of aGVHD. Error bars display means plus or minus SEM (n = 6).

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Additional file 2:

Alloreactive donor T cell infiltrates cause severe tissue damage in acute graft-versus-host disease (aGVHD) target organs. Immunohistochemical stainings of representative small (A) and large (B) bowel samples on day +6 after hematopoietic cell transplantation (HCT) showed a massive infiltration by donor T cells in the major histocompatibility complex (MHC) mismatch model as compared to minor histocompatibility antigen (miHAg) mismatch and syngeneic recipients’ tissues. By day +21, donor T cells infiltrated small and large bowel of the miHAg mismatch transplanted mice and were more abundant than in the organs of syngeneic recipients. Immunofluorescence microscopy of target tissues on day +21 after HCT in the miHAg mismatched compared to syngeneic recipients confirmed these results ((A) and (B), lower panels). CD4+ (Streptavidin-Alexa546, displayed in blue) as well as CD8+ (Alexa-488, displayed in green) donor T cells (CD90.1-APC, displayed in red) massively infiltrated small and large bowel of miHAg mismatch recipients. Double-positive cells appear in purple (CD90.1+ CD4+) and yellow (CD90.1+ CD8+), respectively. In syngeneic recipients, double-positive donor T cells also migrated into these organs, but unlike in allogeneic recipients did not cause tissue damage. (C) Quantification of infiltrating donor T cells showed that more cells infiltrated the small and large bowel in allogeneic than in syngeneic recipients on day +6 (left panel) as well as on day +21 (right panel) after allo-HCT. Similar results were seen in liver and skin. One of three independent experiments is shown (n = 3). Error bars display means plus or minus SEM. HPF, high power field.

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Additional file 3:

There is less donor T cell infiltration in acute graft-versus-host disease (aGVHD) target organs in rapamycin-treated mice than in aGVHD control animals. On day +11 after minor histocompatibility antigen (miHAg) mismatch allogeneic hematopoietic cell transplantation (allo-HCT), donor T cells (CD90.1-APC, displayed in green) massively infiltrated small (A) and large bowel (B) in aGVHD and vehicle control animals whereas the gastrointestinal tract of rapamycin-treated mice showed less donor cell infiltration. Similar results were seen in liver (C) and skin (D) samples. Images show immunofluorescence stainings of one representative mouse per group (n = 5). Cell nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (displayed in white). Images were taken with 40 × magnification. LB, large bowel; SB, small bowel.

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