Identification of a Novel Cancer-Testis Antigen CRT2 Frequently Expressed in Various Cancers Using Representational Differential Analysis
Identification of a Novel Cancer-Testis Antigen CRT2 Frequently Expressed in Various Cancers Using Representational Differential Analysis
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DOI：
10.1158/1078-0432.ccr-07-1374
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发表时间：
2007-11
影响因子：
11.5
通讯作者：
Emiko Hayashi;Yuriko Matsuzaki;Go Hasegawa;T. Yaguchi;S. Kurihara;T. Fujita;T. Kageshita;M. Sano;Y. Kawakami
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文献类型：
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作者：
Emiko Hayashi;Yuriko Matsuzaki;Go Hasegawa;T. Yaguchi;S. Kurihara;T. Fujita;T. Kageshita;M. Sano;Y. Kawakami
目的：睾丸癌抗原是肿瘤免疫治疗的重要靶点。使用代表性差异分析（RDA）和免疫原性评估，试图鉴定在各种癌症中频繁表达的其他癌睾丸抗原。实验设计：用RDA富集睾丸中优先表达的cdna，在睾丸和正常组织之间进行减法。采用逆转录- pcr技术对30个具有癌睾丸样表达的克隆进行评价。鉴定出一种潜在抗原CRT2，并用新产生的抗CRT2抗体分析其表达。采用Western blot和ELISA分析，通过与肿瘤患者血清免疫球蛋白G （IgG）的反应性，以及体外诱导HLA-A24转基因小鼠和人外周血淋巴细胞的肿瘤反应性ctl，检测CRT2的免疫原性。结果：CRT2在正常组织睾丸细长精细胞及各种癌细胞系和组织中均有表达。Western blot和ELISA分析表明，重组CRT2蛋白可被多种肿瘤患者血清IgG识别。一种crt2衍生肽被鉴定为hla - a24限制性t细胞表位，可诱导肿瘤反应性ctl。结论：CRT2是一种新的癌睾丸抗原，表达于睾丸细长精细胞和肿瘤组织（尤其是黑色素瘤）中，可被肿瘤患者血清IgG识别。鉴定出一种能够诱导肿瘤反应性ctl的hla - a24限制性t细胞表位，这表明CRT2可能对癌症诊断和免疫治疗有用。
Purpose: Cancer-testis antigens are promising targets for cancer immunotherapy. Identification of additional cancer-testis antigens with frequent expression in various cancers was attempted using representational differential analysis (RDA) and immunogenicity evaluation. Experimental Design: cDNAs preferentially expressed in testis were enriched using RDA by subtraction between testis and normal tissues. Thirty clones showing cancer-testis–like expression based on EST database analysis were evaluated by reverse transcription-PCR. A potential antigen, CRT2, was identified and its expression was analyzed with a newly generated anti-CRT2 antibody. The immunogenicity of CRT2 was examined based on reactivity with serum immunoglobulin G (IgG) from cancer patients, using Western blot and ELISA analysis, and on in vitro induction of tumor-reactive CTLs from HLA-A24 transgenic mice and human peripheral blood lymphocytes. Results: CRT2 was expressed in elongated spermatids of testis among normal tissues and in various cancer cell lines and tissues. The recombinant CRT2 protein was recognized by serum IgG from patients with various cancers in Western blot and ELISA analyses. A CRT2-derived peptide was identified as an HLA-A24–restricted T-cell epitope that induced tumor-reactive CTLs. Conclusion: CRT2 was identified as a new cancer-testis antigen expressed in elongated spermatids of testis and in cancer tissues (particularly melanoma) that is recognized by serum IgG from cancer patients. An HLA-A24–restricted T-cell epitope capable of inducing tumor-reactive CTLs was identified, suggesting that CRT2 may be useful for cancer diagnosis and immunotherapy.