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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.

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

  • 实验方案

  • 发表日期：

    2025年

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

摘要

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.