Effects of DNA synthesis inhibitors on early antigen expression following primary infection or superinfection by Epstein-Barr virus

Abstract

Seven lymphoid cell lines previously characterized with respect to their resident Epstein-Barr virus (EBV) genome content were infected or superinfected with concentrated EBV from supernatant of the P3HR-1 cell line. Immunofluorescence assays were conducted on smears 48 hours after infection, using human sera containing antibodies to EBV early antigen (EA). Two EBV nuclear antigen (EBNA) negative cell lines containing no detectable resident EBV DNA and five EBNA positive cell lines containing EBV genomes were tested. The cell lines did not spontaneously express EBV EA (i.e., they were non-producers). All cell lines responded to infection or superinfection with EBV by expressing EA. Treatment of the cell lines with arabinosylcytosine (Ara-C) 10 micrograms/ml, at the time of infection resulted in significant decreases in the number of cells expressing detectable EA after drug treatment in all cell lines (72 +/- 5 percent inhibition of EA expression). Experiments were also conducted with hydroxyurea (HU) and phosphonoacetic acid (PAA). It was found that treatment with HU (100 micrograms/ml) inhibited EA production in cell lines containing EBV genome copies by 81 percent as compared to the superinfected cultures receiving no drug. In primary infection of EBNA negative cell lines, HU had minimal effects. PAA (100 micrograms/ml), on the other hand, had very little effect on EA expression following superinfection of cell lines harboring the EBV genome, but reduced the EA expression after primary infection of EBNA negative cell lines by 70 to 80 percent. All drugs were used at concentrations having little effect on RNA and protein synthesis. However, HU and Ara-C significantly reduced DNA synthesis and cell division in the treated cultures.