Construction and characterization of monoclonal antibodies specific for the R transactivator 185 of Epstein-Barr virus

Abstract

Epstein-Barr virus (EBV) has been implicated in the pathogenesis of several human malignancies including B lymphomas and nasopharyngeal carcinoma. The EBV R transactivator (Rta) has been found to play essential roles in stimulating a lytic cycle and viral gene expression. Recently, it was shown that ELISA detecting serum IgG-Rta(150+185) (two internal fragments of Rta) levels may be useful as a serological parameter to assist in the diagnosis of nasopharyngeal carcinoma. The present studies were to prepare monoclonal antibodies specific for the Rta185 and provide a useful tool for the detection of Rta. For this purpose, two monoclonal antibodies (Mabs) specific for the Rta185 were generated. They were identified by Western blot, enzyme-linked immunosorbent assay (ELISA) and immunofluorescence analysis. The results revealed two different immunofluorescence patterns in EBV-positive B cells and epithelial cells, and suggested that there might be a difference in EBV replication mode between B cells and epithelial cells. The Mabs obtained in this study have a potential for the diagnosis of EBV associated diseases.