Recognition of Epstein-Barr virus-associated gastric carcinoma cells by cytotoxic T lymphocytes induced in vitro with autologous lymphoblastoid cell line and LMP2-derived, HLA-A24-restricted 9-mer peptide.
Epstein-Barr virus (EBV) is associated with several types of malignancies including Burkitt's lymphoma, Hodgkin's disease, nasopharyngeal carcinoma, and gastric carcinoma. Previous reports have suggested that EBV-related antigen-targeting immunotherapy is one of the promising approaches for the treatment of these malignancies other than gastric carcinoma. EBV-associated gastric carcinoma (EBVaGC) has been shown to express Epstein-Barr virus nuclear antigen 1 (EBNA1) and latent membrane protein 2 (LMP2). In the present study, DNA and mRNA freshly isolated from tumors of patients with gastric cancer were subjected to polymerase chain reaction (PCR) using EBV-specific primers and reverse transcription (RT)-PCR specific for LMP2 transcripts. EBV-specific region was identified in genomic DNA isolated from cancerous tissues in 22% of gastric cancer patients. LMP2 mRNA was also detected in 3 out of these 5 DNA positive samples tested. To investigate the feasibility of specific immunotherapy for EBVaGC, we induced cytotoxic T lymphocytes (CTLs) from peripheral blood lymphocytes using two kinds of antigen-presenting cells (APCs) such as autologous lymphoblastoid cell line (LCL) and LMP2-derived peptide-pulsed dendritic cells (DCs). The cytotoxicity of these CTLs against peptide-pulsed targets was examined by standard 51Cr release assay and interferon (IFN)-gamma production assay. We further assessed the recognition of tumor cells endogenously expressing LMP2 by these T cells. T cells induced by peptide-loaded DCs and autologous LCL efficiently lysed peptide-pulsed targets. Furthermore, these T cells could recognize not only tumor cells transfected with LMP2, but also LMP2-positive gastric cancer cells which were successfully isolated and cultured from specimens obtained by surgery. Collectively, sensitization of peripheral blood lymphocytes with LMP2-derived peptide was able to induce CTL response against EBVaGC cells. Thus, EBVaGC is susceptible for the LMP2-targeting immunotherapy.
Okugawa
Kaori
K
Department of Surgery, Division of Digestive Surgery, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
Itoh
Tsuyoshi
T
Kawashima
Ichiro
I
Takesako
Kazutoh
K
Mazda
Osam
O
Nukaya
Ikuei
I
Yano
Yutaro
Y
Yamamoto
Yoshiki
Y
Yamagishi
Hisakazu
H
Ueda
Yuji
Y
D000368:Aged; D000369:Aged, 80 and over; D000938:Antigen-Presenting Cells; D002857:Chromium / Q000378:metabolism; D004273:DNA, Neoplasm; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D020031:Epstein-Barr Virus Infections / Q000628:therapy*; D005260:Female; D015234:HLA-A Antigens / Q000276:immunology; D059871:HLA-A24 Antigen; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000378:metabolism; D008214:Lymphocytes / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008875:Middle Aged; D010446:Peptide Fragments / Q000276:immunology; D012334:RNA, Neoplasm; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013274:Stomach Neoplasms / Q000209:etiology* / Q000276:immunology / Q000628:therapy; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014407:Tumor Cells, Cultured; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*
D004273:DNA, Neoplasm; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D015234:HLA-A Antigens; D059871:HLA-A24 Antigen; D010446:Peptide Fragments; D012334:RNA, Neoplasm; D014763:Viral Matrix Proteins; D002857:Chromium; D007371:Interferon-gamma