Dendritic cells adenovirally-transduced with full-length mesothelin cDNA elicit mesothelin-specific cytotoxicity against pancreatic cancer cell lines in vitro
Dendritic cells adenovirally-transduced with full-length mesothelin cDNA elicit mesothelin-specific cytotoxicity against pancreatic cancer cell lines in vitro
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DOI：
10.1016/j.canlet.2011.02.013
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发表时间：
2011-06-01
期刊：
影响因子：
9.7
通讯作者：
Yamaue, Hiroki
中科院分区：
文献类型：
--
作者：
Miyazawa, Motoki;Iwahashi, Makoto;Yamaue, Hiroki
间皮素（MSLN）是胰腺癌免疫治疗分子靶点的一个有吸引力的候选者。本研究的目的是证明，通过表达 MSLN 的转基因树突状细胞 (DC) 刺激，从外周血单核细胞 (PBMC) 产生的细胞毒性 T 淋巴细胞 (CTL) 可以产生针对内源表达 MSLN 的胰腺癌细胞的特异性抗肿瘤免疫。 MSLN 特异性 CTL 是通过用全长 MSLN 基因 (DC-AxCAMSLN) 腺病毒转导的 DC 进行体外刺激，从健康供体的 PBMC 中产生的。使用 4 小时 Cr-51 释放测定法测试细胞毒性活性。使用胰腺癌细胞系（PK1、CfPAC1、AsPC1）、转导 MSLN 基因的类淋巴母细胞系（LCL）和用 MSLN 表位肽脉冲的 LCL 作为靶细胞。 DC-AxCAMSLN 体外刺激诱导的 MSLN 特异性 CTL 杀死了以 HLA 限制方式表达 MSLN 的胰腺癌细胞系。这些 CTL 还表现出针对用多个 MSLN 衍生表位肽脉冲的自体 LCL 的细胞毒活性。此外，从这些CTL中分选的CD8(+) T细胞和CD4(+) T细胞在用DC-AxCAMSLN刺激时显示出显着的干扰素-γ产生。用转有全长 MSLN 基因的 DC 体外刺激 PBMC，通过识别多个 MSLN 表位并激活 CD8(+) T 细胞和 CD4(+) 辅助 T 细胞，引发针对内源表达 MSLN 的胰腺癌细胞系的强效 MSLN 特异性细胞毒活性。因此，这些结果表明使用表达 MSLN 的转基因树突状细胞开发疫苗未来临床应用的潜力。 (C) 2011 Elsevier Ireland Ltd. 保留所有权利。
Mesothelin (MSLN) is an attractive candidate as a molecular target for pancreatic cancer immunotherapy. The purpose of this study was to demonstrate that cytotoxic T lymphocytes (CTLs) generated from peripheral blood mononuclear cells (PBMCs) by stimulation with genetically-modified dendritic cells (DCs) expressing MSLN could produce specific anti-tumor immunity against pancreatic cancer cells endogenously expressing MSLN. MSLN-specific CTLs were generated from PBMCs of healthy donors by in vitro stimulation with DCs adenovirally-transduced with the full-length MSLN gene (DC-AxCAMSLN). The cytotoxic activity was tested using a 4-h Cr-51-release assay. The pancreatic cancer cell lines (PK1, CfPAC1, AsPC1), a lymphoblastoid cell lines (LCL) transduced with the MSLN gene, and LCL pulsed with MSLN-epitope peptides were used as target cells. MSLN-specific CTLs induced by in vitro stimulation with DC-AxCAMSLN killed pancreatic cancer cell lines expressing MSLN in an HLA-restricted fashion. These CTLs also showed cytotoxic activity against autologous LCL pulsed with multiple MSLN-derived epitope peptides. In addition, CD8(+) T cells, as well as CD4(+) T cells, sorted from these CTLs showed significant production of interferon-gamma when stimulated with DC-AxCAMSLN. The in vitro stimulation of PBMCs with DCs transduced with the full-length MSLN gene elicited a potent MSLN-specific cytotoxic activity against pancreatic cancer cell lines endogenously expressing MSLN by recognizing multiple MSLN epitopes and activating both CD8(+) T cells and CD4(+) helper T cells. These results therefore suggest the potential of developing future clinical applications of the vaccines using genetically-modified DCs expressing MSLN. (C) 2011 Elsevier Ireland Ltd. All rights reserved.