Eichmüller Stefan, Usener Dirk, Jochim Anita, Schadendorf Dirk
German Cancer Research Center (DKFZ), Skin Cancer Unit (D0900), Im Neuenheimer Feld 280, Heidelberg, Germany.
Exp Dermatol. 2002 Aug;11(4):292-301. doi: 10.1034/j.1600-0625.2002.110402.x.
Tumor-associated antigens (TAA) are increasingly used as specific targets for immune therapy of malignant melanoma. The aim of the present study was to provide a basis for selecting the most suitable TAA by analyzing the mRNA expression of a large panel of TAA by RT-PCR and Northern blotting. We have chosen primers differentiating four groups of TAA (MAGE-A, MAGE-B, and two groups of GAGE-genes) and 13 individual TAA (MAGE-A2 and -A3, RAGE-1, -2, -3, and -4, LAGE-1a and -1b, NY-ESO-1, GAGE-1, SSX-2, SCP-1, and cTAGE-1) based on most recent sequence data. In addition, the RAGE-gene family has been separated into its four members by a novel designed nested PCR, which was confirmed by Northern analysis. Furthermore, the chromosomal organization and relationship between the RAGE-family and MOK was analyzed. RAGE-4 mRNA could be shown for the first time to be present in testis tissue. The most frequently expressed TAA were the MAGE-A and the GAGE-3,-4,-5,-6,-8 group, whereas among individual TAA MAGE-A2, -A3, RAGE-1, -3, and LAGE-1a/b were found within most specimens and are thus promising candidates for immune therapy. In comparison, melanoma metastatic specimens and cell lines show similar profiles of TAA expression, but individual TAA differ notably between both types of samples indicating that results from cell lines are not always applicable to tumor specimen.
肿瘤相关抗原(TAA)越来越多地被用作恶性黑色素瘤免疫治疗的特异性靶点。本研究的目的是通过逆转录聚合酶链反应(RT-PCR)和Northern印迹法分析大量TAA的mRNA表达,为选择最合适的TAA提供依据。我们根据最新的序列数据选择了能区分四组TAA(MAGE-A、MAGE-B和两组GAGE基因)以及13种个体TAA(MAGE-A2和-A3、RAGE-1、-2、-3和-4、LAGE-1a和-1b、NY-ESO-1、GAGE-1、SSX-2、SCP-1和cTAGE-1)的引物。此外,通过新设计的巢式PCR将RAGE基因家族分离为其四个成员,并通过Northern分析进行了证实。此外,还分析了RAGE家族与MOK之间的染色体组织和关系。首次发现RAGE-4 mRNA存在于睾丸组织中。最常表达的TAA是MAGE-A和GAGE-3、-4、-5、-6、-8组,而在个体TAA中,MAGE-A2、-A3、RAGE-1、-3和LAGE-1a/b在大多数标本中都有发现,因此是免疫治疗的有希望的候选者。相比之下,黑色素瘤转移标本和细胞系显示出相似的TAA表达谱,但两种类型的样本中个体TAA存在显著差异,这表明细胞系的结果并不总是适用于肿瘤标本。