Early Events of Fusion Between Epstein Barr Virus and Human Lymphoblastoid Cells (Raji) Detected by R18 Fluorescence Dequenching Measurements

Abstract

Relief of fluorescence self-quenching was used to monitor fusion (14) of Epstein Barr virus (EBV) with Raji cells after exposure of the virus to a variety of experimental conditions such as neutral or low pH, enzymatic modification of the viral spike glycoproteins, or inhibition of the protein kinase C (PKC) activity. Incubation of the virus at pH 5.9 prior to the binding to the cell membrane led to a significant enhancement of fusion with the plasma membrane. Treatment of Raji cells with an agent known to elevate the endosomal and lysosomal pH (lysosomotropic agent) (3, 12) partially prevented fusion at neutral pH. Desialylation of EBV significantly reduced the extent of fusion with Raji cells. Protein kinase C inhibitor reduced EBV fusion with Raji cells, while treatment with the tumor promoter and the PKC activator TPA caused an increase in the final extent of fusion. Our results suggest that EBV fuses with lymphoblastoid cells in the endocytic vescicles after being rapidly internalized and that protein kinase C is involved in the process of viral entry into cells.

Key Words:

• fusion
• EBV
• fluorescence