Identification of ribosomal protein L19 as a novel tumor antigen recognized by autologous cytotoxic T lymphocytes in lung adenocarcinoma
Identification of ribosomal protein L19 as a novel tumor antigen recognized by autologous cytotoxic T lymphocytes in lung adenocarcinoma
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DOI：
10.1111/j.1349-7006.2009.01351.x
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发表时间：
2010-01-01
期刊：
影响因子：
5.7
通讯作者：
Yasumoto, Kosei
中科院分区：
文献类型：
--
作者：
Kuroda, Koji;Takenoyama, Mitsuhiro;Yasumoto, Kosei
本研究的目的是鉴定一种能够在肺癌患者中诱导特异性细胞免疫反应的新型肿瘤特异性抗原。区域淋巴结淋巴细胞与 CD80 转染的自体肺腺癌细胞系 H1224L 共培养，成功诱导大量细胞毒性 T 淋巴细胞 (CTL)。 CTL 克隆 L7/8 是通过有限稀释法从这些大量 CTL 和裂解的 H1224L 中建立的，但不是自体 Epstein-Barr 病毒转化的 B 细胞或 K562。 CTL 克隆还以 HLA-A*31012 限制的方式识别同种异体肺癌细胞系。利用 CTL 克隆，利用 cDNA 表达克隆技术鉴定出抗原编码基因，该基因编码核糖体蛋白 L19 (RPL19)。最后，通过构建小基因，鉴定出9聚体抗原肽。 RPL19在患者H1224的肺癌组织中过度表达。所有检查的正常组织中 RPL19 mRNA 的表达水平均低于肺癌组织。通过免疫组织化学染色，还发现 RPL19 在 30 个非小细胞肺癌组织中的 12 个（40%）中过表达。肿瘤细胞系中 RPL19 的表达水平与 CTL 克隆 L7/8 响应此类细胞系产生的干扰素 (IFN)-γ 呈正相关。此外，通过转染小干扰RNA抑制RPL19表达，导致cyclinD1、D3合成受到抑制，并抑制过表达RPL19的肺癌细胞系的生长。因此，这种生长抑制可归因于细胞周期的抑制。这些结果可能表明 RPL19 是一种新型过表达抗原，因此可能是作为特异性免疫治疗靶点的有用候选者。 （癌症科学 2009 年）。
The purpose of the present study was to identify a novel tumor-specific antigen capable of inducing a specific cellular immune response in lung cancer patients. The co-culture of regional lymph node lymphocytes and the CD80-transfected autologous lung adenocarcinoma cell line H1224L resulted in a successful induction of bulk cytotoxic T lymphocytes (CTL). CTL clone L7/8 was established by the limiting dilution method from these bulk CTLs and lysed H1224L but not autologous Epstein-Barr virus-transformed B cells or K562. The CTL clone also recognized allogeneic lung cancer cell lines in an HLA-A*31012-restricted manner. Using the CTL clone, an antigen-coding gene was identified using the cDNA expression cloning technique, which encodes ribosomal protein L19 (RPL19). Finally, a 9 mer antigenic peptide was identified by means of construction of mini-genes. RPL19 was overexpressed in the lung cancer tissue from patient H1224. All of the normal tissues examined expressed lower levels of RPL19 mRNA than that of the lung cancer tissue. RPL19 was also found to be overexpressed in 12 of 30 (40%) non-small-cell lung cancer tissues by immunohistochemical staining. The expression level of RPL19 in tumor cell lines correlated positively with the production of interferon (IFN)-gamma by CTL clone L7/8 in response to such cell lines. In addition, the suppression of RPL19 expression by transfection with small interfering RNA resulted in the suppression of cyclinD1, D3 synthesis, and the growth inhibition of lung cancer cell lines overexpressing RPL19. Therefore, this growth suppression could be ascribed to the inhibition of the cell cycle. These results may indicate that RPL19 is a novel overexpressed antigen which may therefore be a useful candidate as a target for specific immunotherapy. (Cancer Sci 2009).