title: Humanized Chronic Graft-versus-Host Disease in NOD-SCID il2rγ-/- (NSG) Mice with G-CSF-Mobilized Peripheral Blood Mononuclear Cells following Cyclophosphamide and Total Body Irradiation.
abstract: Chronic graft-versus-host disease (cGvHD) is the major source of late phase morbidity and mortality after allogeneic hematopoietic stem cell transplantation. Humanized acute GvHD (aGvHD) in vivo models using NOD-SCID il2rγ-/- (NSG) mice are well described and are important tools for investigating pathogenicity of human cells in vivo. However, there have been only few reported humanized cGvHD mouse models. We evaluated if prolonged inflammation driven by low dose G-CSF-mobilized human PBMCs (G-hPBMCs) would lead to cGvHD following cyclophosphamide (CTX) administration and total body irradiation (TBI) in NSG mice. Engraftment was assessed in peripheral blood (PB) and in specific target organs by either flow cytometry or immunohistochemistry (IHC). Tissue samples were harvested 56 days post transplantation and were evaluated by a pathologist. Some mice were kept for up to 84 days to evaluate the degree of fibrosis. Mice that received CTX at 20mg/kg did not show aGvHD with stable expansion of human CD45+ CD3+ T-cells in PB (mean; 5.8 to 23.2%). The pathology and fibrosis scores in the lung and the liver were significantly increased with aggregation of T-cells and hCD68+ macrophages. There was a correlation between liver pathology score and the percentage of hCD68+ cells, suggesting the role of macrophage in fibrogenesis in NSG mice. In order to study long-term survival, 6/9 mice who survived more than 56 days showed increased fibrosis in the lung and liver at the endpoint, which suggests the infiltrating hCD68+ macrophages may be pathogenic. It was shown that the combination of CTX and TBI with a low number of G-hPBMCs (1x106) leads to chronic lung and liver inflammation driven by a high infiltration of human macrophage and mature human T cells from the graft, resulting in fibrosis of lung and liver in NSG mice. In conclusion this model may serve as an important pre-clinical model to further current understanding of the roles of human macrophages in cGvHD.
authors.affiliation: Hematology/Oncology/BMT, The Hospital for Sick Children, Toronto, ON, Canada; Developmental Stem Cell Biology, The Hospital for Sick Children, Toronto, ON, Canada.
authors.forename: Hisaki
authors.initials: H
authors.lastname: Fujii
authors.affiliation: Developmental Stem Cell Biology, The Hospital for Sick Children, Toronto, ON, Canada.
authors.forename: Zhi-Juan
authors.initials: ZJ
authors.lastname: Luo
authors.affiliation: Developmental Stem Cell Biology, The Hospital for Sick Children, Toronto, ON, Canada.
authors.forename: Hye Jin
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Toronto Centre for Phenogenomics, Toronto, ON, Canada.
authors.forename: Susan
authors.initials: S
authors.lastname: Newbigging
authors.affiliation: Hematology/Oncology/BMT, The Hospital for Sick Children, Toronto, ON, Canada.
authors.forename: Adam
authors.initials: A
authors.lastname: Gassas
authors.affiliation: Princess Margaret Hospital, Toronto, ON, Canada.
authors.forename: Armand
authors.initials: A
authors.lastname: Keating
authors.affiliation: Hematology/Oncology/BMT, The Hospital for Sick Children, Toronto, ON, Canada; Developmental Stem Cell Biology, The Hospital for Sick Children, Toronto, ON, Canada.
authors.forename: R Maarten
authors.initials: RM
authors.lastname: Egeler
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D001652:Bile Ducts / Q000187:drug effects / Q000473:pathology; D002908:Chronic Disease; D003520:Cyclophosphamide / Q000494:pharmacology*; D005355:Fibrosis; D006086:Graft vs Host Disease / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D019650:Hematopoietic Stem Cell Mobilization; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007249:Inflammation / Q000473:pathology; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology* / Q000637:transplantation*; D008099:Liver / Q000187:drug effects / Q000473:pathology; D008168:Lung / Q000187:drug effects / Q000473:pathology; D008264:Macrophages / Q000187:drug effects; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009928:Organ Specificity / Q000187:drug effects; D013601:T-Lymphocytes / Q000187:drug effects; D014916:Whole-Body Irradiation; D014945:Wound Healing / Q000187:drug effects
chemical_list: D018952:Antigens, CD34; D016179:Granulocyte Colony-Stimulating Factor; D003520:Cyclophosphamide