Flow cytometry assessment of reactive T‐cells distinguishes classic Hodgkin lymphoma from benign lymphadenopathy in children

BACKGROUND: Detection of classic Hodgkin lymphoma (cHL) neoplastic cells using flow cytometric immunophenotyping (FCI) remains limited. We hypothesized that characterization of the reactive infiltrates could assist in diagnosing cHL in children. METHODS: FCI using four‐color staining approaches was performed on 156 lymph node specimens with the following histopathologic diagnoses: cHL (25 cases), reactive lymphoid hyperplasia (RLH, 44 cases), and non‐Hodgkin lymphoma (87 cases). RESULTS: The overall concordance of FCI data with the histopathologic results of these cases was 81.4%. A reactive expansion of T‐cells with increased expression of CD45RO was present in the reactive infiltrate of cHL (CD45RO/CD3, 67.5%) and Epstein–Barr virus (EBV) infected RLH (62.7%) but not in EBV‐negative RLH (28.0%). The mean fluorescence intensity (MFI) of CD7 was higher for cHL and differed significantly from EBV‐positive RLH (138.5 vs. 63.8). A proposed diagnostic algorithm markedly elevated the overall concordance rate from 81.4% to 97.4%. CONCLUSIONS: Immunophenotyping the reactive infiltrate of lymphoid tissue using flow cytometry is a reliable supplement to histopathology for the rapid diagnosis of pediatric cHL.