2.2.2.2 GvHD Gating SOP

Graft vs. Host Disease data courtesy of Ryan Brinkman’s lab

Data Description

Acute graft versus host disease (GvHD) is one of the most significant clinical problems in allogeneic blood and marrow transplantation. The combination of flow Cytometry, modern bioinformatics, and statistical techniques gives the power to search for patterns of cells in the peripheral blood that correlate with a variety of physiologic or disease states including GvHD.

Goal is to look for populations that correlate to GvHD development, peripheral blood samples from patients undergoing allogeneic blood and marrow transplant are analyzed for 121 subpopulations defined by 4-color, 6-parameter combinations of 25 phenotypic and activation markers. Patient samples are stained with 25 antibodies in 4-color combinations.

Once the populations of interest are isolated by gating, their number as a proportion of peripheral blood mononuclear cells is calculated. These values are compared to results from the same patient over time.

Ryan Brinkman, et al. determined that proportion of CD3⁺ CD4⁺ CD8β⁺ cells post transplant did correlate with development of GvHD. Biology of Blood and Marrow Transplantation 13:691-700 (2007) PDF

GvHD SOP

Your Goal is to select the CD3+ CD4+ CD8β+ cells in 12 samples which correspond to timepoints for 1 patient. Files are available either on the proviced thumbdrives or in Illinois/FlowDx/GvHD_TATSIS.zip. You will only have the panel 2 samples in the workspace GvHDrun1TATSIS.wsp. Please adjust the gates to be as accurate as you would normally gate these populations. If you want to use different types of gates to get to the final populations, please create your own gating tree, but also adjust the existing one. Lymphocytes T cells CD4+, CD8b+ Save your workspace with this nomenclature: GvHDrun1_yourname.wsp

Preliminary Results of the GvHD Gate-a-thon