A preclinical acute GVHD mouse model based on chemotherapy conditioning and MHC-matched transplantation

Abstract

Animal disease models have been criticized for lack of resemblance to human illnesses, hampering transfer of knowledge from preclinical research to clinical medicine. In the field of allogeneic hematopoietic stem cell transplantation (allo-HSCT), it is standard practice to study GVHD in lethal TBI-based murine models. Frequently, MHC-mismatched donors are used in GVHD models. In contrast, in clinical allo-HSCT conditioning with chemotherapy (+/ − TBI) is common and donors are often MHC-matched. Aiming at a more clinically oriented situation, we established and characterized a murine MHC-matched, minor histocompatibility antigen mismatched GVHD model (LP/J [H2k b ] → C57BL/6 [H2k b ]) using busulfan and cyclophosphamide conditioning. We found typical clinical and histological features of acute GVHD. T-cell infiltration, GVHD-specific damage and systemic inflammation were similar to observations made in patients after allo-HSCT. In survivors of acute GVHD, we found expansion of CD4+ T cells and the development of scleroderma-like chronic GVHD. The use of chemotherapy-based, minor histocompatibility antigen (miHA)-mismatched GVHD animal models may be a good option when studying clinically relevant questions in the field of allo-HSCT.

The Role of Animal Models in the Study of Hematopoietic Stem Cell Transplantation and GvHD: A Historical Overview

“…This model was developed to more closely mimic the clinical situation, where patients usually are MHC matched. With this model, similar T cell infiltration, GvHD-specific damage, and systemic inflammation were observed in the mice as reported in humans (273). Thus, animal models of selective mHA mismatch may represent human HSCT more closely than MHC-mismatched models (193).…”

The Green Tea Catechin Epigallocatechin Gallate Ameliorates Graft-versus-Host Disease

“…To analyze target organ damage we a minor mismatch GVHD model (LP→B6) [16] and applied histopathological scores after Lerner et al [17] At day +17, we found significantly reduced histopathological GVHD scores in liver, colon and skin in EGCG treated allo-BMT recipients vs. controls without EGCG treatment (Fig 3A). In the liver, untreated control allo-BMT recipients had GVHD-associated massively increased periportal infiltration of leukocytes whereas in EGCG treated allo-BMT recipients, infiltration was only slightly increased (Fig 3B).…”

“…Both models have been described previously. [9, 16] Allo-BMT recipients received a combination of 25mg/kg EGCG (Enzo Life Sciences, Lörrach, Germany) and 1mg/kg quercetin (Sigma Aldrich) dissolved in DMSO (20μl total) and PBS i.p. daily or EGCG and quercetin alone (for clinical score) or DMSO only (20μl, control) respectively from day 0 to end of experiment.…”

“…[16] Single cell suspensions were stained with rat anti-mouse antibodies from BD Biosciences as follows: CD3e-APC-Cy7 (1:100), CD4-PE-Cy7 (1:800), CD8a-APC (1:200), CD25-PerCP-Cy5.5(1:200), CD11b-APC-Cy7 (1:400), B220-PerCP-Cy5.5 (1:100) or Nk1.1-PerCP-Cy5.5 (1:200). For chimerism analysis of blood and BM, antibodies against PE- Ly9.1 (1:100) and FITC-H2kb (1:50) were used.…”

Conditioning with Fludarabine-Busulfan versus Busulfan-Cyclophosphamide Is Associated with Lower aGVHD and Higher Survival but More Extensive and Long Standing Bone Marrow Damage

“…Compared with the publication of Sadeghi et al [14], in which all mice died within 60 days (median survival = 11 days) after transplantation, the Bu-Cy conditioning regimen in our study resulted in more uniform survival of aGVHD mice. Of note, Riesner et al [24] have showed a delayed aGVHD in a murine MHC-matched, miHA-mismatched model [LP/J (H2k b ) →C57BL/6(H2k b )] using Bu (100 mg/kg)-Cy (300 mg/kg) conditioning. These discrepancies between our study and previous publications may be due to the degree of allomismatch (MHC-mismatched or miHA-mismatched), differences in graft (bone marrow cells and splenocytes), and the intensity of conditioning.…”