Use of an in vitro immunoselected tumor line to identify shared melanoma antigens recognized by HLA-A*0201-restricted T cells.
Use of an in vitro immunoselected tumor line to identify shared melanoma antigens recognized by HLA-A*0201-restricted T cells.
复制标题
DOI：
--
复制
发表时间：
2001-02
期刊：
影响因子：
11.2
通讯作者：
M. Harada;Yong F Li;M. El-Gamil;Steven A. Rosenberg;P. Robbins
中科院分区：
文献类型：
--
作者：
M. Harada;Yong F Li;M. El-Gamil;Steven A. Rosenberg;P. Robbins
免疫选择的黑色素瘤细胞系已失去了显性黑色素瘤抗原MART-1和gp 100的表达，试图确定以前未知的肿瘤抗原。在用自体免疫选择肿瘤系重复刺激后，从单个黑素瘤患者的外周血建立了许多HLA-A*0201限制性T细胞克隆。一个T细胞克隆（C-22）识别16个HLA-A2+黑素瘤细胞系中的14个，以及HLA-A2+黑素细胞，但既不识别HLA-A2+成纤维细胞也不识别自体B细胞。自体cDNA文库的筛选导致与表达序列标签数据库中的条目相同的转录物的分离。北方印迹分析表明，该基因在大多数黑色素瘤细胞系和黑素细胞中表达，而在正常组织中不表达。基于对截短的cDNA的识别和使用共有HLAA*0201结合基序的研究，鉴定了克隆C-22识别的肽表位（AMF-GREFCYA）。发现第二个T细胞克隆（C-29）以HLA-A*0201限制性方式识别新的酪氨酸酶相关蛋白2表位（455-463; YAIDLPVSV）。总之，这些结果提供了可用于开发HLA-A2+黑色素瘤患者的免疫方案的额外靶点，并证明了免疫选择肿瘤系用于鉴定新黑色素瘤抗原的实用性。
An immunoselected melanoma cell line that had lost expression of the dominant melanoma antigens MART-1 and gp100 was generated in an attempt to identify previously unknown tumor antigens. After repeated stimulation with the autologous immunoselected tumor line, a number of HLA-A*0201-restricted T-cell clones were established from the peripheral blood of a single melanoma patient. One T-cell clone (C-22) recognized 14 of 16 HLA-A2+ melanoma cell lines, as well as HLA-A2+ melanocytes but recognized neither HLA-A2+ fibroblasts nor autologous B cells. Screening of an autologous cDNA library resulted in the isolation of a transcript identical to an entry in the expressed sequence tag database. Northern blot analysis revealed that this gene was expressed in most melanoma cell lines and melanocytes but not in normal tissues. The peptide epitope (AMF-GREFCYA) recognized by clone C-22 was identified based on studies of the recognition of truncated cDNAs and the use of the consensus HLAA*0201 binding motif. A second T-cell clone (C-29) was found to recognize a new tyrosinase-related protein 2 epitope (455-463; YAIDLPVSV) in an HLA-A*0201-restricted manner. Together, these results provide additional targets that can be used for the development of immunotherapeutic protocols in HLA-A2+ melanoma patients and demonstrate the utility of immunoselected tumor lines for the identification of new melanoma antigens.