Epstein-Barr virus (EBV)-carrying and -expressing T-cell lines established from severe chronic active EBV infection.
Four novel Epstein-Barr virus (EBV)-carrying T-cell lines, designated SIS, AIK-T8, AIK-T4, and SKN, were established from peripheral blood lymphocytes (PBL) of patients with severe chronic active EBV infection, in the presence of interleukin-2 and 4-deoxyphorbol ester. AIK-T8 and -T4 were derived from a single patient. Cell marker and genotype analyses showed that SIS, AIK-T8, and AIK-T4 had mature T-cell phenotypes with clonally rearranged T-cell receptor (TCR) genes, whereas SKN had an immature T-cell phenotype without TCR gene rearrangement. None of the cell lines expressed B, natural killer, or myeloid antigens or had Ig gene rearrangement. All lines carried EBV genomes in a single episomal form. SIS, AIK-T8, and SKN showed the same phenotype, TCR gene configuration, and/or EBV clonotype as their source or biopsied materials; therefore, they represented EBV-infected T cells proliferating in the patients. TCR gene and EBV episomal structures similar to those of AIK-T4 were not found in its source PBL, probably due to the few parental clones in vivo. All lines expressed EBV-encoded small RNA (EBER) 1, nuclear antigen (EBNA) 1, and latent membrane protein (LMP) 1, -2A, and -2B, but not other EBNAs that could be recognized by EBV-specific immune T cells. EBV replicative antigens were rarely expressed or induced. Such EBV latency reflects the in vivo situation, in which the T cells may evade immune surveillance and be insensitive to antiherpesvirus drugs. Collectively, the data suggest that EBV can target and latently infect T cells at any stage of differentiation in vivo, thus potentially causing uncontrolled T-cell proliferation. These cell lines will facilitate further analyses of possible EBV-induced oncogenicity in T cells.
Imai
S
S
Department of Virology, Hokkaido University School of Medicine, Sapporo, Japan.
Sugiura
M
M
Oikawa
O
O
Koizumi
S
S
Hirao
M
M
Kimura
H
H
Hayashibara
H
H
Terai
N
N
Tsutsumi
H
H
Oda
T
T
Chiba
S
S
Osato
T
T
D001483:Base Sequence; D002460:Cell Line; D002472:Cell Transformation, Viral; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D015967:Gene Expression Regulation, Viral; D015329:Gene Rearrangement, T-Lymphocyte; D006566:Herpesviridae Infections / Q000473:pathology*; D004854:Herpesvirus 4, Human; D006801:Humans; D007223:Infant; D008297:Male; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics; D013601:T-Lymphocytes / Q000382:microbiology*; D014412:Tumor Virus Infections / Q000473:pathology*
D017931:DNA Primers; D004279:DNA, Viral; D012333:RNA, Messenger; D016693:Receptors, Antigen, T-Cell, alpha-beta