CD21-mediated entry and stable infection by Epstein-Barr virus in canine and rat cells.
We developed an adenovirus vector for transduction of the human CD21 gene (Adv-CD21), the Epstein-Barr virus (EBV)-specific receptor on human B lymphocytes, to overcome the initial barrier of EBV infection in nonprimate mammalian cells. Inoculation of Adv-CD21 followed by exposure to recombinant EBV carrying a selectable marker resulted in the successful entry of EBV into three of seven nonprimate mammalian cell lines as evidenced by expression of EBV-determined nuclear antigen (EBNA). The EBV-susceptible cell lines included rat glioma-derived 9L, rat mammary carcinoma-derived c-SST-2, and canine kidney-derived MDCK. Subsequent selection culture with G418 yielded drug-resistant cell clones. In these cell clones, EBV existed as an episomal form, as evidenced through the Gardella gel technique. Among the known EBV latency-associated gene products, EBV-encoded small RNAs, EBNA1 and transcripts from the BamHI-A rightward reading frame (BARF0), and latent membrane protein 2A were expressed in all EBV-infected cell clones. The viral lytic events could be induced in these cell clones by simultaneous treatment with 12-O-tetradecanoylphorbol-13-acetate and n-butyric acid, but they were abortive, and infectious virus was not produced. These results indicate that once the initial barrier for attachment is overcome artificially, EBV can establish a stable infection in some nonprimate mammalian cells, and they raise the possibility that transgenic animals with the human CD21 gene could provide an animal model for EBV infection.
Yang
L
L
Department of Tumor Virology, Institute for Genetic Medicine, Hokkaido University, Kita-ku, Sapporo 060-8638, Japan.
Maruo
S
S
Takada
K
K
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D006224:Cricetinae; D004279:DNA, Viral / Q000737:chemistry; D004285:Dogs; D005822:Genetic Vectors; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D010957:Plasmids / Q000235:genetics; D051381:Rats; D017464:Receptors, Complement 3d / Q000235:genetics / Q000502:physiology*; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction
D004279:DNA, Viral; D017464:Receptors, Complement 3d