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JoVE Journal

Immunology and Infection

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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

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06:06 min

May 23rd, 2019

May 23rd, 2019

6,873 Views

0:04

Title

0:40

Non-Obese Diabetic (NOD) Scid Gamma (NSG) Mouse Irradiation and Peripheral Blood Mononuclear Cell (PBMC) Injection

2:22

Retro-Orbital Injection

3:24

Clinical Graft Versus Host Disease (GVHD) Scoring

3:58

Genomic DNA Isolation and Human T Cell Quantification

4:59

Results: Representative GVHD Disease Progression and Digital PCR Human T Cell Detection

5:37

Conclusion

Transcribir

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

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Here, we present a protocol to induce and score disease in a xenogeneic graft-versus-host disease (xenoGVHD) model. xenoGVHD provides an in vivo model to study immunosuppression of human T cells. Additionally, we describe how to detect human T cells in tissues with digital PCR as a tool to quantify immunosuppression.

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