A large group of juvenile Hodgkin's disease patients (n = 242, mean age 11.7 years, 75% [n = 181] seropositive) was evaluated for anti-Epstein-Barr virus (EBV) antibody responses and the presence of EBV-encoded EBER-RNA and latent membrane protein-1 (LMP1)-protein expression in the tumor. The molecular diversity of anti-EBV antibody responses in Hodgkin's disease patients with EBV-positive and-negative tumors was studied by enzyme-linked immunosorbent assay (ELISA) and immunoblot. Using purified recombinant LMP1 protein as antigen, the presence of antibodies to LMP1 was related to expression of LMP1 in the tumor cells and specific EBV-serological patterns. Antibodies to LMP1 were detected in 30% of the EBV-seropositive Hodgkin's disease patients. The presence of antibodies to LMP1 was not associated with a distinct anti-EBV antibody diversity profile (ELISA), but a significantly higher percentage of patients with antibodies to LMP1 had antibodies to ZEBRA and viral capsid antigen (VCA)-p18 (Immunoblot). Significantly more patients with an EBV-positive tumor had detectable antibody responses to LMP1, but the presence of antibodies to LMP1 did not reflect the expression of LMP1 protein in the tumor cells. Interestingly, all patients with the strongest antibody responses to LMP1 had EBV-negative tumors, suggesting immunological selection in vivo.
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