Induction and Scoring of Graft-Versus-Host Disease in a Xenogeneic Murine Model and Quantification of Human T Cells in Mouse Tissues using Digital PCR

This protocol outlines how to induce xenograft-versus-host disease, or xenoGVHD, and how to blind and standardize clinical scoring to ensure consistent results. The xenoGVHD model provides an in vivo method for testing immunosuppressive therapies against human rather than murine T cells, improving the translation of these studies to the clinic. Demonstrating the procedure will be Amara Seng, a graduate student from my laboratory.

One day before the injection, place eight-to 12-week-old, non-obese diabetic scid gamma, or NSG, mice in a sterilized pie cage, and irradiate the mice in a cesium-137 source with a total dose of 150 centigrays, with a slow rotation to ensure an even irradiation. Then, place the mice into clean cages in a sterile biosafety cabinet overnight. The next morning, collect enough healthy human blood to isolate 1.1 times 10 to the seven peripheral blood mononuclear cells, or PBMCs, per mouse, and dilute the heparinized blood in an equal volume of 2%fetal bovine serum, or FBS, in PBS.