Full-length EBNA1 mRNA-transduced dendritic cells stimulate cytotoxic T lymphocytes recognizing a novel HLA-Cw*0303- and -Cw*0304-restricted epitope on EBNA1-expressing cells.
Epstein-Barr virus (EBV)-encoded nuclear antigen 1 (EBNA1) is an attractive target for immunotherapy against EBV-associated malignancies because it is expressed in all EBV-positive cells. Although CD8+ cytotoxic T-lymphocyte (CTL) epitope presentation is largely prevented by its glycine-alanine-repeat domain (GAr), the use of mRNA-transduced dendritic cells (DCs) would offer the advantage of priming EBNA1-specific CTLs. After stimulation with GAr-containing EBNA1-transduced monocyte-derived DCs, two EBNA1-specific CTL clones, B5 and C6, were isolated successfully from a healthy donor. These CTLs recognize peptides in the context of HLA-B*3501 and HLA-Cw*0303, respectively. A novel epitope, FVYGGSKTSL, was then identified, presented by both HLA-Cw*0303 and -Cw*0304, which are expressed by >35% of Japanese, >20% of Northern Han Chinese and >25% of Caucasians. The mixed lymphocyte-peptide culture method revealed that FVYGGSKTSL-specific CTL-precursor frequencies in HLA-Cw*0303- or -Cw*0304-positive donors were between 1x10(-5) and 1x10(-4) CD8+ T cells. Moreover, both CTL clones inhibited growth of HLA-matched EBV-transformed B lymphocytes in vitro, and B5 CTLs produced a gamma interferon response to EBNA1-expressing gastric carcinoma cells in the context of HLA-Cw*0303. These data demonstrate that EBNA1 mRNA-transduced DCs may be useful tools for inducing EBNA1-specific CTLs that might be of clinical interest for CTL therapy of EBV-associated malignancies.
Ito
Yoshinori
Y
Division of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.
Demachi-Okamura
Ayako
A
Division of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.
Ohta
Rieko
R
Division of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.
Akatsuka
Yoshiki
Y
Division of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.
Nishida
Keiko
K
Division of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.
Tsujimura
Kunio
K
Division of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.
Morishima
Yasuo
Y
Department of Cell Therapy, Aichi Cancer Center Hospital, Nagoya, Japan.
Takahashi
Toshitada
T
Division of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.
Kuzushima
Kiyotaka
K
Division of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.
D000483:Alleles; D001402:B-Lymphocytes / Q000276:immunology; D049109:Cell Proliferation; D003713:Dendritic Cells / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics / Q000276:immunology*; D015236:HLA-C Antigens / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014170:Transformation, Genetic
D018984:Epitopes, T-Lymphocyte; D019309:Epstein-Barr Virus Nuclear Antigens; D015236:HLA-C Antigens; C106548:HLA-C*03:04 antigen; D007371:Interferon-gamma; C417029:EBV-encoded nuclear antigen 1