title: Clinical value of the flow cytometric method for measuring lymphocyte subset activation: spontaneous activation of T-cell subpopulations is associated with acute GvHD.
abstract: Thymidine ((3)H-TdR) incorporation remains the most commonly used method to quantifying T-cell proliferation. This method, however, does not provide information about specific lymphocyte subpopulations responding to different stimuli. In our study, we modified previously described nonradioactive flow-cytometric T-cell activation assay measuring the expression of a CD69+ antigen on T-cell subsets and applied it to analysis of lymphocyte subsets activation/proliferation in children after allogeneic hematopoietic cell transplantation (HCT). We compared the percentage of spontaneously activated lymphocyte subpopulations (background) and the percentage of PHA-P, PWM, and SEB-stimulated cell subsets from two groups of patients: group 1, children with Graft versus Host Disease (GvHD) and group 2, children without any signs of GvHD at the time of analysis. High rate of spontaneous T-cell subset activation was found in group 1 with CD3+CD8+Ts cells being the most affected cell population. High background activation of Th and B cells correlated with the occurrence of autoimmune phenomena posttransplant. Rapid quantification of CD69+ expression on unstimulated and stimulated T-cell subsets proved to be a valuable method for monitoring children after allogeneic HCT. High proportion of activated, unstimulated Ts cells observed in the GvHD group may underline the critical role of CD3+CD8+ cells in the pathogenesis of GvHD. Thus in future immunosuppressive therapy may be adjusted according to the proportion of activated Ts cells.
authors.affiliation: Department of Pediatric Hematology and Oncology, Medical University of Wroclaw, Wroclaw, Poland. kk@pedhemat.am.wroc.pl
authors.forename: K
authors.initials: K
authors.lastname: Kalwak
authors.forename: D
authors.initials: D
authors.lastname: Turkiewicz
authors.forename: M
authors.initials: M
authors.lastname: Ussowicz
authors.forename: E
authors.initials: E
authors.lastname: Gorczyńska
authors.forename: J
authors.initials: J
authors.lastname: Toporski
authors.forename: R
authors.initials: R
authors.lastname: Ryczan
authors.forename: B
authors.initials: B
authors.lastname: Rybka
authors.forename: D
authors.initials: D
authors.lastname: Noworolska-Sauren
authors.forename: A
authors.initials: A
authors.lastname: Chybicka
mesh_terms: D016026:Bone Marrow Transplantation / Q000276:immunology; D002648:Child; D005434:Flow Cytometry / Q000379:methods; D005500:Follow-Up Studies; D006086:Graft vs Host Disease / Q000276:immunology*; D006801:Humans; D007938:Leukemia / Q000145:classification / Q000601:surgery / Q000628:therapy; D008213:Lymphocyte Activation; D008212:Lymphocyte Depletion; D016131:Lymphocyte Subsets / Q000276:immunology*; D009190:Myelodysplastic Syndromes / Q000601:surgery / Q000628:therapy; D033581:Stem Cell Transplantation; D013601:T-Lymphocytes / Q000276:immunology*; D013936:Thymidine / Q000378:metabolism; D013997:Time Factors; D014184:Transplantation, Homologous / Q000276:immunology*; D014316:Tritium
chemical_list: D014316:Tritium; D013936:Thymidine