Although multiparameter-flow-cytometry (MFC) has demonstrated clinical relevance in MGUS/myeloma, immunophenotypic studies on the full spectrum of Waldenström's macroglobulinemia (WM) remain scanty. Herein, a comprehensive MFC analysis on BM samples from 244 newly diagnosed patients with an IgM monoclonal protein was performed, including 67 IgM-MGUS, 77 smoldering, and 100 symptomatic WM. Our results show a progressive increase on the number and light-chain-isotype-positive B-cells from IgM-MGUS to smoldering and symptomatic WM (P<0.001), with only 1% of IgM-MGUS patients showing >10% B-cells or 100% light-chain-isotype-positive B-cells (P<0.001). Complete light-chain restriction of the B-cell compartment was an independent prognostic factor for time-to progression in smoldering WM (median 26 months; HR:19.8, P=0.001) and overall survival in symptomatic WM (median 44 months; HR:2.6, P=0.004). The progressive accumulation of light-chain-isotype-positive B-cells accompanied the emergence of a characteristic Waldenstrom's phenotype (CD22(+dim)/CD25(+)/CD27(+)/IgM(+)) that differed from other B-NHL by negative expression of CD5, CD10, CD11c or CD103. In contrast to myeloma, light-chain-isotype-positive plasma cells in IgM monoclonal gammopathies show otherwise normal antigenic expression. Our results highlight the potential value of MFC immunophenotyping for the characterization of the Waldenström's clone, as well as for the differential diagnosis, risk of progression and survival in WM.Leukemia accepted article preview online, 22 April 2013; doi:10.1038/leu.2013.124.