title: High-content flow cytometry and temporal data analysis for defining a cellular signature of graft-versus-host disease.
abstract: Acute graft-versus-host disease (GVHD) is diagnosed by clinical and histologic criteria that are often nonspecific and typically apparent only after the disease is well established. Because GvHD is mediated by donor T cells and other immune effector cells, we sought to determine whether changes within a wide array of peripheral blood lymphocyte populations could predict the development of GvHD. Peripheral blood samples from 31 patients undergoing allogeneic blood and marrow transplant were analyzed for the proportion of 121 different subpopulations defined by 4-color combinations of lymphocyte phenotypic and activation markers at progressive time points posttransplant. Samples were processed using a newly developed high content flow cytometry technique and subjected to a spline- and functional linear discriminant analysis (FLDA)-based temporal analysis technique. This strategy identified a consistent posttransplant increase in the proportion and extent of fluctuation of CD3+CD4+CD8beta+ cells in patients who developed GVHD compared to those that did not. Although larger prospective clinical studies will be necessary to validate these results, this study demonstrates that high-content flow cytometry coupled with temporal analysis is a powerful approach for developing new diagnostic tools, and may be useful for developing a sensitive and specific predictive test for GVHD.
authors.affiliation: Terry Fox Laboratory/British Columbia Cancer Agency, Vancouver, Canada.
authors.forename: Ryan Remy
authors.initials: RR
authors.lastname: Brinkman
authors.forename: Maura
authors.initials: M
authors.lastname: Gasparetto
authors.forename: Shang-Jung Jessica
authors.initials: SJ
authors.lastname: Lee
authors.forename: Albert J
authors.initials: AJ
authors.lastname: Ribickas
authors.forename: Janelle
authors.initials: J
authors.lastname: Perkins
authors.forename: William
authors.initials: W
authors.lastname: Janssen
authors.forename: Renee
authors.initials: R
authors.lastname: Smiley
authors.forename: Clay
authors.initials: C
authors.lastname: Smith
mesh_terms: D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; D005434:Flow Cytometry / Q000379:methods*; D006086:Graft vs Host Disease / Q000175:diagnosis*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects; D016130:Immunophenotyping / Q000379:methods; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000473:pathology*; D011237:Predictive Value of Tests; D013995:Time
chemical_list: D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; C477867:CD8beta antigen