| Autor: |
Battiwalla, Minoo, Hepgur, Mehmet, Pan, Dalin, McCarthy, Philip L., Ahluwalia, Manmeet S., Camacho, Susan H., Starostik, Petr, Wallace, Paul K. |
| Zdroj: |
Cytometry Part B: Clinical Cytometry; September 2010, Vol. 78 Issue: 5 p348-356, 9p |
| Abstrakt: |
Background:Glycosylphosphatidylinositol GPInegative blood cells are diagnostic for Paroxysmal Nocturnal Hemoglobinuria PNH. Marrow failure states are often associated with GPInegative cell populations. Quantification of small clonal populations of GPInegative cells influences clinical decisions to administer immunosuppressive therapy in marrow failure states aplastic anemia or myelodysplastic syndrome and to monitor minimal residual disease after allogeneic blood or marrow transplantation BMT. We studied the reliability of highresolution flow cytometry markers operating at the limits of detection.Methods:We performed serial quantification of the PNH clone size in 38 samples using multiparameter flow cytometry. Granulocytes, monocytes, and RBCs were gated using forward and side scatter as well as lineagespecific markers. The GPIlinked markers fluorescent aerolysin FLAER, CD55, and CD59 were comparatively evaluated. We also evaluated CD16 on granulocytes and CD14 on monocytes. The sensitivity of detection by each marker was further defined by serial dilution experiments on a flowsorted sample. Two patients had quantification of their GPInegative clones before and after allogeneic BMT.Results:FLAER was the most discriminant marker and allowed identification of 0.1 of GPInegative cells despite other markers having superior signaltonoise characteristics. CD14 and CD16 were inferior to CD55 at lower concentrations and in clinical application.Conclusions:Multiparameter flow cytometry permits quantification of small GPInegative clones with a sensitivity limit of about 0.1. The single most reliable marker to monitor small granulocyte or monocyte PNH clones is FLAER, especially in conditions such as myelodysplastic syndromes or BMT, when traditional GPIlinked surface marker expression can be significantly altered. © 2010 International Clinical Cytometry Society |
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