Demonstration by single-cell PCR that Reed-Sternberg cells and bystander Blymphocytes are infected by different Epstein-Barr virus strains in Hodgkin's disease

Epstein-Barr virus (EBV) is associated with Hodgkin's disease (HD). However , EBV-positive Reed-Sternberg (RS) cells and EBV-positive B lymphocytes co- exist in the same EBV-positive lymph node affected by HD. In a previous rep ort, using total lymph node DNA, the presence of two distinct EBV strains w as demonstrated, but their cellular localization (i.e. RS cells vs B lympho cytes) could not be determined. To address this question, three patients wi th EBV-associated HD were selected in the present study and single-cell PCR of the latent membrane protein-1 (LMP-1) gene from isolated RS cells was p erformed. In one case, it was clear that RS cells and B lymphocytes were in fected by different EBV strains. In the two remaining cases, only one band was detected from total lymph node DNA. However, single-cell PCR showed tha t RS cells in each sample were infected by single EBV strains, which were d ifferent from those detected in lymphoblastoid cell lines derived from EBV- positive B lymphocytes of lymph node cell suspensions from these two patien ts.