Modelling Graft-Versus-Host Disease in Mice Using Human Peripheral Blood Mononuclear Cells

Manjurul Haque; Dominic A Boardman; Avery J Lam; Katherine N MacDonald; Lieke Sanderink; Qing Huang; Vivian C W Fung; Sabine Ivison; Majid Mojibian; Megan K Levings

doi:10.21769/BioProtoc.4566

Modelling Graft-Versus-Host Disease in Mice Using Human Peripheral Blood Mononuclear Cells

Bio Protoc. 2022 Dec 5;12(23):e4566. doi: 10.21769/BioProtoc.4566.

Authors

Manjurul Haque^{1

2}, Dominic A Boardman^{1

2}, Avery J Lam^{1

2}, Katherine N MacDonald^{2

3}, Lieke Sanderink^{1

2}, Qing Huang^{1

2}, Vivian C W Fung^{1

2}, Sabine Ivison^{1

2}, Majid Mojibian^{1

2}, Megan K Levings^{1

2

3}

Affiliations

¹ Department of Surgery, The University of British Columbia, Vancouver BC, Canada.
² BC Children's Hospital Research Institute, Vancouver BC, Canada.
³ School of Biomedical Engineering, The University of British Columbia, Vancouver BC, Canada.

Abstract

Graft-versus-host disease (GvHD) is a significant complication of allogeneic hematopoietic stem cell transplantation. In order to develop new therapeutic approaches, there is a need to recapitulate GvHD effects in pre-clinical, in vivo systems, such as mouse and humanized mouse models. In humanized mouse models of GvHD, mice are reconstituted with human immune cells, which become activated by xenogeneic (xeno) stimuli, causing a multi-system disorder known as xenoGvHD. Testing the ability of new therapies to prevent or delay the development of xenoGvHD is often used as pre-clinical, proof-of-concept data, creating the need for standardized methodology to induce, monitor, and report xenoGvHD. Here, we describe detailed methods for how to induce xenoGvHD by injecting human peripheral blood mononuclear cells into immunodeficient NOD SCID gamma mice. We provide comprehensive details on methods for human T cell preparation and injection, mouse monitoring, data collection, interpretation, and reporting. Additionally, we provide an example of the potential utility of the xenoGvHD model to assess the biological activity of a regulatory T-cell therapy. Use of this protocol will allow better standardization of this model and comparison of datasets across different studies. Graft-versus-host disease (GvHD) is a significant complication of allogeneic hematopoietic stem cell transplantation. In order to develop new therapeutic approaches, there is a need to recapitulate GvHD effects in pre-clinical, in vivo systems, such as mouse and humanized mouse models. In humanized mouse models of GvHD, mice are reconstituted with human immune cells, which become activated by xenogeneic (xeno) stimuli, causing a multi-system disorder known as xenoGvHD. Testing the ability of new therapies to prevent or delay the development of xenoGvHD is often used as pre-clinical, proof-of-concept data, creating the need for standardized methodology to induce, monitor, and report xenoGvHD. Here, we describe detailed methods for how to induce xenoGvHD by injecting human peripheral blood mononuclear cells into immunodeficient NOD SCID gamma mice. We provide comprehensive details on methods for human T cell preparation and injection, mouse monitoring, data collection, interpretation, and reporting. Additionally, we provide an example of the potential utility of the xenoGvHD model to assess the biological activity of a regulatory T-cell therapy. Use of this protocol will allow better standardization of this model and comparison of datasets across different studies. This protocol was validated in: Sci Transl Med (2020), DOI: 10.1126/scitranslmed.aaz3866 Graphical abstract.

Keywords: Graft-versus-host disease (GvHD); Hematopoietic stem cell transplantation (HSCT); Regulatory T cell; Thymic T-regulatory cells; Xenograft.