Flow Cytometric Evaluation of Double/Triple Hit Lymphoma

Oncol Res. 2016;23(3):137-46. doi: 10.3727/096504015X14500972666761.


"Double" or "triple" hit lymphomas (D/THL) with recurrent translocations involving MYC/8q24 and BCL2/18q21 and/or BCL6/3q27 are characterized by a poor prognosis, but their identification is hampered by the clinicopathologic overlap with other disease categories. Cases with circulating blastic-appearing cells may initially cause concern for lymphoblastic leukemia a diagnostic dilemma, which has not been well studied. There is only limited literature regarding the flow cytometric (FC) D/THL phenotype and its clinical correlates. The FC features of 20 D/THL (11 BCL2(+)/MYC(+), 5 BCL6(+)/MYC(+), 4 BCL2(+)/BCL6(+)/MYC(+)) were evaluated, compared to 20 B-lymphoblastic leukemias (B-LBL), and correlated with overall survival. Most (89%, 17/19) D/THL were CD10(+), 47% (9/19) lacked surface light chain, and a significant subset underexpressed CD45 (47%, 9/19), CD20 (42% 8/19), and/or CD19 (39%, 7/18), which did not vary by genetic subgroup. Compared to B-LBL, D/THL less frequently underexpressed CD45 (p = 0.0001) and CD20 (p = 0.0004). Lower levels of BCL2 expression were noted in the BCL6(+)/MYC(+) and BCL2(+)/BCL6(+)/MYC(+) subgroups versus BCL2(+)/MYC(+) cases (p = 0.0014). Of the flow cytometric parameters assessed, dim CD45 expression correlated with inferior survival (p = 0.01). Although there is some overlap with B-LBL, D/THL demonstrates a characteristic immunophenotype which may have prognostic significance and warrants further investigation.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Antigens, CD19 / immunology
  • Antigens, CD20 / immunology
  • Female
  • Flow Cytometry*
  • Gene Expression Regulation, Neoplastic / immunology
  • Humans
  • Immunophenotyping*
  • Leukocyte Common Antigens / immunology
  • Male
  • Middle Aged
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / diagnosis
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / genetics*
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / immunology
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / pathology
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / immunology
  • Proto-Oncogene Proteins c-bcl-6 / genetics
  • Proto-Oncogene Proteins c-bcl-6 / immunology
  • Proto-Oncogene Proteins c-myc / genetics
  • Proto-Oncogene Proteins c-myc / immunology
  • Translocation, Genetic / genetics*


  • Antigens, CD19
  • Antigens, CD20
  • BCL6 protein, human
  • CD19 molecule, human
  • MYC protein, human
  • Proto-Oncogene Proteins c-bcl-2
  • Proto-Oncogene Proteins c-bcl-6
  • Proto-Oncogene Proteins c-myc
  • Leukocyte Common Antigens
  • PTPRC protein, human