Evaluation of CD229 as a new alternative plasma cell gating marker in the flow cytometric immunophenotyping of monoclonal gammopathies

Abstract

BACKGROUND: Current flow-cytometric plasma cell (PC) gating is based on CD138, CD38, and CD45 expression. CD138 is known for variable expression and loss during storage and processing. Introduction of anti-CD38 and anti-CD138 monoclonal-antibody therapies has limited the use of these markers during follow-up. Hence, additional reliable PC-gating markers are required. Recently, CD229 has been claimed as an alternative PC-gating marker. However, these studies are limited to a small cohort of samples. We evaluated the utility of CD229 as a new PC-gating marker in routine laboratory practice. METHODS: Expression of CD229 was studied in 310 bone marrow (BM) samples (251 plasma-cell disorders and 59 controls) and compared with CD138 and CD38 expression. We also evaluated the effect of additional processing for cytoplasmic immunoglobulin-light-chains (CyIgL) staining on the quantitation of PC. RESULTS: Expression of CD229 was consistently stronger on PC than other hematopoietic-cells (p < 0.001). PC-percentages using CD229 in combination with CD38 or CD138 and CD45 revealed high correlation with a reference gating-strategy using CD138, CD38 and CD45 (r = 0.98, r = 0.99 r = 0.99 respectively) and r = 0.92 using CD229 and CD45 without CD38 or CD138. In contrast, CD138 expression showed significant variability (CV-MFI, 97.5) and loss from PC in 53% of samples. Quantitation of PC was found to be lower in 69.3% and higher in 30.7% samples processed for CyIgL-staining as compared to surface-staining. CONCLUSIONS: CD229 is a reliable new alternative PC-gating marker in routine laboratory practice. Quantitation of PC based on CD138 expression or from samples processed for CyIgL-staining should be used with caution. © 2018 International Clinical Cytometry Society.