bims-hummad 2025-03-23 papers

Methods Mol Biol. 2025 ;2907 183-206

Humanized Mouse Model of Graft-Versus-Host Disease Utilizing Nonirradiated NOD-scid-IL-2Rγnull Mice.

Amal Elhage, Chloe Sligar, Debbie Watson, Ronald Sluyter.

Humanized mouse models of graft-versus-host disease (GVHD), involving the injection of human immune cells into immunodeficient mice, are firmly established and offer avenues for exploring pathways implicated in GVHD development and for testing therapeutics to prevent or treat this disease. NOD-scid-IL-2Rγnull (NSG) mice are a strain of immunodeficient mice commonly used to study GVHD. This chapter details methods relating to this model including human immune cell isolation, the injection of these human immune cells into nonirradiated NSG mice, monitoring the mice for signs of clinical GVHD (including weight loss), immunolabeling with anti-human and anti-mouse monoclonal antibodies of leukocytes isolated from the spleens, livers, and lungs of humanized mice, evaluating human cell engraftment and human and mouse immune cell subsets via flow cytometry and measuring serum cytokine concentrations using a multiplexed flow cytometric assay.

Keywords: Allogeneic hematopoietic stem cell transplantation; Cytokine; Flow cytometry; Human peripheral blood mononuclear cell; Humanized mouse model; Leukocyte; NSG mouse; Xenogeneic graft-versus-host disease

DOI: https://doi.org/10.1007/978-1-0716-4430-0_9

Methods Mol Biol. 2025 ;2907 333-358

Use of Human Chronic Myeloid Leukemia Cells to Study Graft-Versus-Leukemia Immunity in a Xenogeneic Mouse Model of Graft-Versus-Host Disease.

Chloe Sligar, Nicolas R Tomasiello, Peter Cuthbertson, Taylor L Nelson, Amal Elhage, Ronald Sluyter, Debbie Watson.

Humanized mouse models of graft-versus-host disease (GVHD) primarily focus on treatments to prevent disease progression. However, there is little investigation into whether graft-versus-leukemia (GVL) immunity is retained following these treatments for GVHD. Here, we describe the methods to generate a humanized mouse model to study GVL immunity against firefly luciferase (luc) expressing human K562 chronic myeloid leukemia cells in humanized NOD-scid-IL2Rγnull mice. This chapter provides an overview of human peripheral blood mononuclear cell isolation, culturing of K562-luc chronic myeloid leukemia cells, injection of these cells into immunodeficient mice, monitoring for signs of disease, tracking of leukemia using an in vivo imaging system, and assessment of human cell engraftment and GVL immunity using flow cytometry.

Keywords: Allogeneic hematopoietic stem cell transplantation; Chronic myeloid leukemia; Flow cytometry; Graft-versus-leukemia immunity; In vivo imaging system; NSG mice; Xenogeneic graft-versus-host disease

DOI: https://doi.org/10.1007/978-1-0716-4430-0_16

Methods Mol Biol. 2025 ;2907 57-70

Xenogeneic Animal Models of Graft-Versus-Host Disease.

Ronald Sluyter, Chloe Sligar, Amal Elhage, Kang Min Chu, Debbie Watson.

Allogeneic hematopoietic stem cell transplantation (HSCT) is used to treat malignant and non-malignant blood disorders, but its efficacy is limited by the development of graft-versus-host disease (GVHD). Thus, better understanding and new biomarkers and therapies are required to combat this disease. Xenogeneic animal models of GVHD help to address these gaps and to translate findings from allogeneic animal models of GVHD and other laboratory findings to human allogeneic HSCT recipients. This chapter discusses different humanized mouse models used to study GVHD. Further, this chapter introduces a humanized rat model of GVHD, as well as other xenogeneic models of GVHD in which immune-deficient mice receive blood cells from dogs, cows, or horses. Collectively, this chapter serves as a brief account of xenogeneic animal models of GVHD.

Keywords: Bovine; Canine; Equine; Human; Leukemia; Leukocytes; Lymphocytes; Murine; Rodent; Xenogeneic graft-versus-host disease

DOI: https://doi.org/10.1007/978-1-0716-4430-0_2