Mouse Models of Graft-Versus-Host Disease (GvHD)

GvHD in human patients | Acute GvHD | Chronic GvHD | Readouts | Results - Acute GvHD | References

GvHD in human patients

Graft-versus-host disease is a serious and common complication of an allogeneic bone marrow or stem cell transplant. The disease is a result of T cells in the donor's graft recognizing the recipient's tissue (host) as foreign, resulting in an immunologic attack of the graft against the host.

There are two types of GvHD:

• Acute GvHD: in humans, generally occurs in less than 100 days from transplantation. Usually involves the skin, gastrointestinal tract, and liver. The most common symptom is a pruritic or painful rash.
• Chronic GvHD: in humans, generally occurs 100 or more days after transplantation. Symptoms include dry eyes or vision changes, skin rash, sores in the mouth and throat, liver dysfunction, gastrointestinal upset, lung damage, muscle weakness, and joint stiffness.

The preventive approach to GvHD often involves pre-transplant treatment with a calcineurin inhibitor (tacrolimus or cyclosporine) and methotrexate, or post-transplant treatment with cyclophosphamide. Topical and systemic steroids are also commonly administered.

Additional FDA-approved treatments include:

• Orencia (abatacept, T cell co-stimulation modulator) for acute GvHD
• Jakafi (ruxolitinib, selective JAK1/2 inhibitor) for steroid-refractory acute and chronic GvHD
• Imbruvica (ibrutinib, Bruton's tyrosine kinase inhibitor) for chronic GvHD
• Rezurock (belumosudil, ROCK2 inhibitor) for chronic GvHD

Mouse models

Acute or chronic GvHD is induced in mice using allogeneic or xenogeneic grafts with or without first irradiating recipient mice.

Acute xenogeneic GvHD in NOG mice

Xenogeneic GvHD using immune-deficient recipient mice with human peripheral blood mononuclear cells (hPBMCs) allows for the in vivo study of human-T-cell-mediated GvHD. In this model, immunodeficient mice are used as recipients to prevent graft rejection.

At Hooke, we use either irradiated or non-irradiated NOG (NOD.Cg-Prkdc^scid Il2rg^tm1Sug/JicTac) recipient mice and hPBMCs. NOG mice lack mature T, B, and NK cells, and have dysfunctional macrophages and dendritic cells.

The severity of GvHD is influenced by the number of cells transferred and the dose of irradiation used to facilitate donor cell engraftment.

Chronic allogeneic GvHD in mice - under development (2023)

Allogeneic GvHD is induced in recipient mice using cells from donor mice with mismatched major histocompatibility complex (MHC) or minor histocompatibility antigens. Recipients may be irradiated before cell transfer to prevent graft rejection.

As of 2023, Hooke is establishing 3 models of chronic GvHD:

• Cells from DBA/2 (H2d) mice in non-irradiated B6D2F1 (H2b/d) recipient mice
• Cells from DBA/2 (H2d) mice in irradiated BALB/c (H2d) recipient mice
• Cells from C57BL/6 (H2b) mice in irradiated BALB/c (H2d) recipient mice

Readouts

Readouts include body weight and GvHD clinical scores. The following measurements and analyses can also be performed:

• Flow cytometric analysis of blood cells
• Anti-dsDNA concentration in plasma or serum
• Histological and flow cytometric analysis of lungs, liver, spleen, skin, colon, and/or small intestine

Results - Acute GvHD

The results below are from a representative study performed at Hooke. NOG recipient mice were either not irradiated or irradiated one day prior to intravenous injection of the indicated number of hPBMCs.

Body weights, clinical scores, and survival curves

Flow cytometry - blood

Flow cytometry was performed on blood collected 7, 14, and 21 days post-cell transfer. The dot plots below are from a representative mouse from the group that was irradiated and one day later injected with 6x10⁶ hPBMCs.

References

Schroeder MA, DiPersio JF. 2011. May;4(3):318-33.

Justiz Vaillant AA, Modi P, Mohammadi O. Graft Versus Host Disease. [Updated 2022 Oct 10]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2023.