Systematic identification of human melanoma antigens using serial analysis of gene expression (SAGE)
Systematic identification of human melanoma antigens using serial analysis of gene expression (SAGE)
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DOI：
10.1097/00002371-200501000-00002
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发表时间：
2005-01-01
影响因子：
3.9
通讯作者：
Kawakami, Y
中科院分区：
文献类型：
--
作者：
Matsuzaki, Y;Hashimoto, S;Kawakami, Y
为了通过系统的基因表达分析结合患者血清的免疫原性筛选来鉴定新的黑色素瘤抗原，采用基因表达序列分析（SAGE）方法对高度着色的黑色素瘤细胞系SKmel23的转录本进行了分析。测序了25,997个SAGE标签，其中包含10,382个独特的转录本。将该黑色素瘤SAGE文库与睾丸SAGE文库和结肠SAGE数据库进行比较，并与黑素细胞cDNA文库随机测序获得的cDNA数据库进行比较。在最终选择的15个标签中，以相对较高的频率优先表达黑色素瘤和黑色素细胞为标准，进一步分析了两个标签的结构和免疫原性。其中一个被鉴定为PAX3，其异构体PAX3d在黑色素瘤和黑色素细胞中主要表达。另一株来源于一个新基因，并分离出其全长cDNA克隆。RT-PCR和Northern blot分析证实了这些基因在黑色素瘤和黑色素细胞中的优先表达。重组细菌PAX3d蛋白在部分黑色素瘤和VKH患者血清IgG中被识别，但在健康人群中未被识别，提示PAX3d是黑色素瘤和VKH患者的一种新的黑色素细胞特异性抗原。作者报告了两种黑色素细胞/黑色素瘤特异性分子，这可能有助于这些色素疾病的诊断或治疗。此外，使用SAGE和患者血清免疫筛选的系统被证明是系统识别肿瘤抗原的有效方法。
To identify new melanoma antigens using systematic gene expression analysis combined with rapid screening of patient sera for immunogenicity, a serial analysis of gene expression (SAGE) method was applied to profile transcripts in a highly pigmented melanoma cell line SKmel23. 25,997 SAGE tags consisting of 10,382 unique transcripts were sequenced. This melanoma SAGE library was compared with a testis SAGE library and the colon SAGE database, and to the cDNA database obtained by random sequencing of a melanocyte cDNA library. Among the 15 tags finally selected with criteria of preferential expression on melanoma and melanocytes at relatively high frequency, two tags were further analyzed for their structure and immunogenicity. One was identified as PAX3, and its isoform, PAX3d, was found to be dominantly expressed in melanoma and melanocytes. The other was derived from a novel gene and its full-length cDNA clone was isolated. Preferential expression of these genes in melanoma and melanocytes was confirmed by RT-PCR and Northern blot analysis. The recombinant bacterial PAX3d protein was recognized by serum IgG from some patients with melanoma and Vogt-Koyanagi-Harada (VKH) disease but not from healthy individuals, indicating that PAX3d is a new melanocyte-specific antigen immunogenic in patients with melanoma or VKH disease. The authors report two melanocyte/melanoma-specific molecules, which may be useful for development of diagnosis or treatment of these pigment disorders. In addition, a system using SAGE and immunoscreening with patients' sera is shown to be an efficient method for the systematic identification of tumor antigens.