Rapid identification and characterization of infected cells in blood during chronic active Epstein-Barr virus infection
Benjamin Fournier, David Boutboul, Julie Bruneau, Charline Miot, Cécile Boulanger, Marion Malphettes, Isabelle Pellier, Bertrand Dunogué, Benjamin Terrier, Felipe Suárez, Stéphane Blanche, Martin Castelle, Sarah Winter, Henri‐Jacques Delecluse, Thierry Jo Molina, Capucine Pïcard, Stephan Ehl, Despina Moshous, Lionel Galicier, Vincent Barlogis, Alain Fischer, Bénédicte Neven, Sylvain Latour
Abstract
Epstein-Barr virus (EBV) preferentially infects epithelial cells and B lymphocytes and sometimes T and NK lymphocytes. Persistence of EBV-infected cells results in severe lymphoproliferative disorders (LPDs). Diagnosis of EBV-driven T or NK cell LPD and chronic active EBV diseases (CAEBV) is difficult, often requiring biopsies. Herein, we report a flow-FISH cytometry assay that detects cells expressing EBV-encoded small RNAs (EBERs), allowing rapid identification of EBV-infected cells among PBMCs. EBV-infected B, T, and/or NK cells were detectable in various LPD conditions. Diagnosis of CAEBV in 22 patients of Caucasian and African origins was established. All exhibited circulating EBV-infected T and/or NK cells, highlighting that CAEBV is not restricted to native American and Asian populations. Proportions of EBV-infected cells correlated with blood EBV loads. We showed that EBV-infected T cells had an effector memory activated phenotype, whereas EBV-infected B cells expressed plasma cell differentiation markers. Thus, this method achieves accurate and unambiguous diagnoses of different forms of EBV-driven LPD and represents a powerful tool to study their pathophysiological mechanisms.