Mitotic EBNA-positive lymphocytes in peripheral blood during infectious mononucleosis

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Abstract
Infectious mononucleosis (IM) is usually a benign lymphoproliferative disease caused by Epstein–Barr virus (EBV)1. Although EBV induces a state of continuous proliferation in infected B lymphocytes in vitro2,3, the most prominent lymphoproliferation during IM is of activated, or atypical, T lymphocytes presumably responding to the virus or virus-infected cells4. However, EBV genome-carrying cells are known to be circulating during IM, as cultured peripheral blood leukocytes from patients with the disease give rise to continuous lymphoblastoid cell lines, each cell of which contains the EBV genome and expresses the EBV determined nuclear antigen (EBNA)5,6. The proposal that EBV-infected cells in IM blood are not endowed with enhanced growth potential but are merely latently infected7 is supported by demonstrations that cells infected in vivo enter a viral replicative cycle when placed in vitro and that most cell lines derived from cultured lymphocytes of IM patients are infected by virus released in vitro8. However the cells could also be capable of proliferation in vivo, since virus production and transformation are not mutually exclusive properties of EBV-transformed cells9. Recently, EBNA has been detected in a very small fraction of peripheral blood lymphocytes of IM patients after T cells were first removed10,11 and this has been interpreted to indicate that cell transformation occurs in vivo during IM. The isolation of colonies of EBNA-positive cells from IM blood leukocytes cultures in soft agar suggests that at least some infected cells are capable of direct outgrowth into transformed cells12. We report here direct evidence that circulating EBV-infected cells exhibit increased growth properties during IM.
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