Immunophenotypic characterization of reactive and neoplastic plasmacytoid dendritic cells permits establishment of a ten-color flow cytometric panel for initial workup and residual disease evaluation of blastic plasmacytoid dendritic cell neoplasm

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare hematopoietic neoplasm whose immunophenotype remains incompletely characterized, particularly with regards to its distinction from reactive plasmacytoid dendritic cells (PDC). This limitation complicates detection of low-level involvement by BPDCN as well as minimal residual disease (MRD) assessment following therapy. We conducted the current study to characterize the immunophenotype of BPDCN in a cohort of 39 patients, and compared it to that of reactive PDC. We found that, in addition to CD56 expression (97%), BPDCN showed a number of aberrancies, including decreased/negative CD38 (82%), positive CD7 (64%), negative CD2 (81%), negative CD303 (56%), increased HLA-DR (69%) and decreased CD123 (78%) expression. Although BPDCN cells were characterized by CD56 expression, reactive PDC consistently included a CD56(+) subset, ranging from 1.3%-20% (median 4.5%) of all PDC, challenging the detection of MRD. These CD56(+) reactive PDC were, however, consistently positive for CD2 and CD303, brightly positive for CD38, and negative for CD7, distinctively different from BPDCN. Based on these findings, we set up a ten-color flow cytometry assay for BPDCN and validated it to a sensitivity of 0.01%. This panel was prospectively tested in 19 bone marrow samples from seven patients with BPDCN, and it effectively distinguished BPDCN cells from background reactive PDC in all cases. In summary, by understanding the immunophenotype of reactive and neoplastic PDC, BPDCN can be effectively detected by flow cytometry to a very low level using a panel of markers in addition to CD56. Such an assay could be used for initial bone marrow workup as well as MRD detection after therapy.