Daniel N Tran1,2,3, Sandy A B C Smith1, David A Brown1,2,4, Andrew J C Parker5, Joanne E Joseph2,4,6, Nicola Armstrong3,7, William A Sewell1,2,3. 1. Immunology Department, SydPath, St Vincent's Hospital Sydney, New South Wales, Australia. 2. St Vincent's Clinical School, University of New South Wales, New South Wales, Australia. 3. Garvan Institute of Medical Research, New South Wales, Australia. 4. Applied Medical Research Centre, St Vincent's Hospital Sydney, New South Wales, Australia. 5. Anatomical Pathology Department, SydPath, St Vincent's Hospital Sydney, New South Wales, Australia. 6. Haematology Department, SydPath, St Vincent's Hospital Sydney, New South Wales, Australia. 7. Mathematics and Statistics, Murdoch University, Western Australia, Australia.
Abstract
BACKGROUND: There is an emerging role for flow cytometry (FC) in the assessment of small populations of plasma cells (PC). However, FC's utility has been questioned due to consistent underestimation of the percentage of PC compared to microscopy. METHODS: A retrospective study was performed on bone marrow samples analysed by 8-colour FC. Plasma cell populations were classified as polyclonal or monoclonal based on FC analysis. FC findings were compared with microscopy of aspirates, histology and immunohistochemistry of trephine biopsies, and immunofixation (IFX) of serum and/or urine. RESULTS: FC underestimated PC compared to aspirate and trephine microscopy. The 10% diagnostic cutoff for MM on aspirate microscopy corresponded to a 3.5% cutoff on FC. Abnormal plasma cell morphology by aspirate microscopy and clonality by FC correlated in 229 of 294 cases (78%). However, in 50 cases, FC demonstrated a monoclonal population but microscopy reported no abnormality. In 15 cases, abnormalities were reported by microscopy but not by FC. Clonality assessment by trephine microscopy and FC agreed in 251/280 cases (90%), but all 29 discordant cases were monoclonal by FC and not monoclonal by microscopy. These cases had fewer PC and proportionally more polyclonal PC, and when IFX detected a paraprotein, it had the same light chain as in the PC determined by FC. CONCLUSIONS: FC was more sensitive in detecting monoclonal populations that were small or accompanied by polyclonal PC. This study supports the inclusion of FC in the evaluation of PC, especially in the assessment of small populations.
BACKGROUND: There is an emerging role for flow cytometry (FC) in the assessment of small populations of plasma cells (PC). However, FC's utility has been questioned due to consistent underestimation of the percentage of PC compared to microscopy. METHODS: A retrospective study was performed on bone marrow samples analysed by 8-colour FC. Plasma cell populations were classified as polyclonal or monoclonal based on FC analysis. FC findings were compared with microscopy of aspirates, histology and immunohistochemistry of trephine biopsies, and immunofixation (IFX) of serum and/or urine. RESULTS: FC underestimated PC compared to aspirate and trephine microscopy. The 10% diagnostic cutoff for MM on aspirate microscopy corresponded to a 3.5% cutoff on FC. Abnormal plasma cell morphology by aspirate microscopy and clonality by FC correlated in 229 of 294 cases (78%). However, in 50 cases, FC demonstrated a monoclonal population but microscopy reported no abnormality. In 15 cases, abnormalities were reported by microscopy but not by FC. Clonality assessment by trephine microscopy and FC agreed in 251/280 cases (90%), but all 29 discordant cases were monoclonal by FC and not monoclonal by microscopy. These cases had fewer PC and proportionally more polyclonal PC, and when IFX detected a paraprotein, it had the same light chain as in the PC determined by FC. CONCLUSIONS: FC was more sensitive in detecting monoclonal populations that were small or accompanied by polyclonal PC. This study supports the inclusion of FC in the evaluation of PC, especially in the assessment of small populations.
Authors: Dominik F Draxler; Lisa M Wutzlhofer; Georg Slavka; Wolfgang Hübl; Heinz Ludwig; Martin Schreder; John Reynolds; Martin Willheim Journal: Indian J Hematol Blood Transfus Date: 2019-03-07 Impact factor: 0.900