Histological Assessment of the Sclerotic Graft-versus-Host Response in the Humanized RAG2−/−γc−/− Mouse Model

Abstract

Graft-versus-host disease (GVHD) remains a frequently occurring and difficult-to-treat complication in human allogeneic stem cell transplantation. Murine transplantation models are often used to study and understand the complex pathogenesis of GVHD and to explore new treatment strategies. Although GVHD kinetics may differ in murine and human models, adequate models are essential for identification of the crucial factors responsible for the major pathology in GVHD. We present a detailed description of the specific histological features of a graft-versus-host-induced fibrotic response in xenogeneic RAG2(-/-)γc(-/-) mice after total body irradiation and injection with human peripheral blood mononuclear cells. We describe the full morphological features of this reaction, including a detailed analysis of the specific tissue infiltration patterns of the human peripheral blood mononuclear cells. Our data show the development of fibrosis, predominantly near blood vessels, and reveal different cell populations and specific cell migration patterns in the affected organs. The combination of immunohistochemical cell characterization and mRNA expression analysis of both human (donor)- and murine (host)-derived cytokines reveals an interaction between host tissues and donor-derived cells in an entangled cytokine profile, in which both donor- and host-derived cytokines contribute to the formation of fibrosis.

Modifying Graft-versus-Host Disease in a Humanized Mouse Model by Targeting Macrophages or B-Cells

“…Mice were monitored for symptoms of GvHD as described previously [28]. Mice were sacrificed by cervical dislocation after 70 days or when losing more than 20% of their original body weight or when they experienced severe symptoms, wound infection of signs of bleeding caused by the given injections.…”

“…Mice were monitored for development of symptoms of GvHD as described previously [28] and sacrificed by cervical dislocation either after 70 days or when either losing more than 20% of their original body weight, having severe symptoms, wound infection, or bleeding because of the given injections or when they experienced side effects of the medication. FACS analysis with a FACSCalibur (Becton Dickinson) was performed to evaluate the fraction of human cells in the murine peripheral blood, using human CD45 (huCD45), mouse CD45, huCD3, and huCD19 (Becton Dickinson), as described by van Rijn et al [26].…”

CD28 Blockade Ex Vivo Induces Alloantigen-Specific Immune Tolerance but Preserves T-Cell Pathogen Reactivity

“…A GvHD score was calculated based on four criteria—posture, activity, fur texture, and skin integrity—each scored 0, 1, or 2, as described ( 31 ). Upon sacrifice, livers were fixed in 4% paraformaldehyde (Sigma-Aldrich), sectioned, stained with hematoxylin (Dako Agilent Technologies, Glostrup, Denmark) and eosin (Sigma-Aldrich) and histologically analyzed for tissue necrosis, vasodilation, and immune cell infiltration ( 32 ). Tissue necrosis was quantified by measuring necrotic areas using Image J software and vasodilation by calculating the mean of the maximum diameters of all vessels in a high power field.…”

Donor T Cells Administered Over HLA Class II Barriers Mediate Antitumor Immunity without Broad Off-Target Toxicity in a NOD/Scid Mouse Model of Acute Leukemia

“…Several in vivo models for xenogeneic GVHD developed in immunodeficient mice through the administration of human PBMCs or purified T cells have been proposed as models for human GVHD as well [38][39][40][41][42]. In these models, however, whether xeno-reactivity as mediated by human T cells sufficiently resembles HLA-restricted alloreactivity in humans remains unknown.…”