NEOPLASIA Molecular single-cell analysis of the clonal relationship of small Epstein-Barr virus–infected cells and Epstein-Barr virus–harboring Hodgkin and Reed/Sternberg cells in Hodgkin disease

Abstract

Epstein-Barr virus (EBV) can be detected in the tumor cells of approximately 40% of cases of classical Hodgkin disease (cHD). Clonality studies suggest that in-fection of the neoplastic Hodgkin and Reed/Sternberg (HRS) cells occurs be-fore tumor clone expansion. In EBV-positive cases, variable numbers of EBER-positive small B cells are sometimes also observed that immunohistologically dif-fer from the neoplastic cells by lack of CD30 and latent membrane protein 1 ex-pression. To analyze the clonal relation-ship between these EBV1 cells and the HRS cells, single EBV-infected CD302 B cells, as well as HRS cells from 3 cases of EBV-positive cHD were micromanipu-lated, their immunoglobulin gene rear-rangements amplified and then com-pared with each other. In 2 cases, all small EBV-infected cells were clonally unre-lated to the HRS cells. In a third case, 2 of 29 small CD302 cells were found to carry HRS cell-specific rearrangements. Thus, small CD302 EBV-infected B cells in cHD belong to the HRS tumor clone rarely, if at all. In all cases, small clones unrelated to the HRS cell clones were identified among the small EBV1 CD302 cells. The vast majority of small EBV1 CD302 B cells was found to carry somatically mutated V re-gion genes, indicating that in lymph nodes of patients with HD, like in the peripheral blood of healthy individuals, EBV per-sists in memory B cells. (Blood. 2000;96

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