Fusion of EBV with the surface of receptor-negative human hepatoma cell line Li7A permits virus penetration and infection.
Our preliminary data suggest that Epstein-Barr virus (EBV) is able to bind to and fuse with the surface membranes of hepatoma cell line Li7A. In order to obtain further evidence, we utilized the relief of rhodamine fluorescence to monitor whether fusion would also take place when Li7A cells were exposed to experimental conditions such as neutral or low pH. It is well known that for some viruses, protonation in the endosomal compartment is needed to trigger the fusion. We show, furthermore, that the rate and extent of fusion are not affected by pretreatment of the cells with agents known to elevate the lysosomal and ensodomal pH, such as chloroquine or NH4Cl (lysosomotropic agent). By indirect immunofluorescence assay, in addition, we confirmed the binding of the EBV to the Li7A cell surface membrane. We attempted finally to correlate the above processes with successful infection of Li7A cells by EBV detected using the polymerase chain reaction technique. In spite of the apparent lack of viral receptor CD21, these nonlymphoid cells appeared susceptible to EBV penetration and infection through fusion with the plasma membrane at the surface of the cells.
Lisi
A
A
Istituto di Medicina Sperimentale C.N.R., Roma, Italy.
Pozzi
D
D
Carloni
G
G
Da Villa
G
G
Iacovacci
S
S
Valli
M B
MB
Grimaldi
S
S
D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D001483:Base Sequence; D002712:Chlorides / Q000494:pharmacology; D002738:Chloroquine / Q000494:pharmacology; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D005434:Flow Cytometry; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008561:Membrane Fusion / Q000187:drug effects; D008969:Molecular Sequence Data; D017672:Nitrogen Compounds / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D002712:Chlorides; D017931:DNA Primers; D004279:DNA, Viral; D017672:Nitrogen Compounds; D014759:Viral Envelope Proteins; C090195:glycoprotein 85, Epstein-Barr virus; D002738:Chloroquine; C037454:nitrogen chloride