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首页> 外文期刊>Journal of immunotherapy >Systematic Identification of Human Melanoma Antigens Using Serial Analysis of Gene Expression (SAGE).
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Systematic Identification of Human Melanoma Antigens Using Serial Analysis of Gene Expression (SAGE).

机译：使用基因表达序列分析（SAGE）系统鉴定人类黑素瘤抗原。

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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.
机译：为了使用系统的基因表达分析结合患者血清的免疫原性快速筛选来鉴定新的黑色素瘤抗原，将基因表达的系列分析（SAGE）方法应用于高度色素沉着的黑色素瘤细胞系SKmel23中的转录本。对由10,382个独特转录本组成的25,997个SAGE标签进行了测序。将该黑素瘤SAGE文库与睾丸SAGE文库和结肠SAGE数据库进行比较，并与通过黑素细胞cDNA文库的随机测序获得的cDNA数据库进行比较。最后以相对较高的频率在黑素瘤和黑素细胞上优先表达的标准选择的15个标签中，进一步分析了两个标签的结构和免疫原性。一种被鉴定为PAX3，发现其同工型PAX3d主要在黑素瘤和黑素细胞中表达。另一个来自新基因，并分离出其全长cDNA克隆。通过RT-PCR和Northern印迹分析证实了这些基因在黑素瘤和黑素细胞中的优先表达。重组细菌PAX3d蛋白被一些黑色素瘤和Vogt-Koyanagi-Harada（VKH）病患者的血清IgG识别，但未被健康个体识别，这表明PAX3d是一种新的黑色素瘤或VKH病患者免疫原性的黑色素特异性抗原。作者报告了两种黑素细胞/黑素瘤特异性分子，可能有助于这些色素紊乱的诊断或治疗。此外，使用SAGE以及对患者血清进行免疫筛查的系统被证明是系统鉴定肿瘤抗原的有效方法。

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