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.
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.
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DOI：
10.3892/or.12.4.725
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发表时间：
2004-10
期刊：
影响因子：
4.2
通讯作者：
K. Okugawa;Tsuyoshi Itoh;I. Kawashima;K. Takesako;O. Mazda;I. Nukaya;Yutaro Yano;Yoshiki Yamamoto;H. Yamagishi;Y. Ueda
中科院分区：
文献类型：
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作者：
K. Okugawa;Tsuyoshi Itoh;I. Kawashima;K. Takesako;O. Mazda;I. Nukaya;Yutaro Yano;Yoshiki Yamamoto;H. Yamagishi;Y. Ueda
EB 病毒 (EBV) 与多种类型的恶性肿瘤有关，包括伯基特淋巴瘤、霍奇金病、鼻咽癌和胃癌。先前的报告表明，EBV 相关抗原靶向免疫疗法是治疗除胃癌以外的这些恶性肿瘤的有前途的方法之一。 EBV 相关胃癌 (EBVaGC) 已被证明表达 Epstein-Barr 病毒核抗原 1 (EBNA1) 和潜伏膜蛋白 2 (LMP2)。在本研究中，使用 EBV 特异性引物和 LMP2 转录物特异性的逆转录 (RT)-PCR，对从胃癌患者肿瘤中新鲜分离的 DNA 和 mRNA 进行聚合酶链反应 (PCR)。从 22% 的胃癌患者的癌组织中分离出的基因组 DNA 中发现了 EBV 特异性区域。在这 5 个 DNA 阳性样本中，有 3 个检测到了 LMP2 mRNA。为了研究 EBVaGC 特异性免疫治疗的可行性，我们使用两种抗原呈递细胞 (APC)（例如自体淋巴母细胞系 (LCL) 和 LMP2 衍生的肽脉冲树突细胞 (DC)）从外周血淋巴细胞诱导细胞毒性 T 淋巴细胞 (CTL)。通过标准 51Cr 释放测定和干扰素 (IFN)-γ 产生测定来检查这些 CTL 针对肽脉冲靶标的细胞毒性。我们进一步评估了这些 T 细胞对内源表达 LMP2 的肿瘤细胞的识别。由负载肽的 DC 和自体 LCL 诱导的 T 细胞可有效裂解肽脉冲的靶标。此外，这些T细胞不仅可以识别转染LMP2的肿瘤细胞，还可以识别从手术获得的标本中成功分离和培养的LMP2阳性胃癌细胞。总的来说，LMP2 衍生肽对外周血淋巴细胞的敏化能够诱导针对 EBVaGC 细胞的 CTL 反应。因此，EBVaGC 对 LMP2 靶向免疫疗法敏感。
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.