微卫星不稳定阳性子宫内膜癌患者对一种癌/睾丸抗原CAGE的频繁免疫反应。

Frequent immune responses to a cancer/testis antigen, CAGE, in patients with microsatellite instability-positive endometrial cancer.

作者信息

Iwata Takashi, Fujita Tomonobu, Hirao Nobumaru, Matsuzaki Yuriko, Okada Tsutomu, Mochimaru Hiroshi, Susumu Nobuyuki, Matsumoto Eri, Sugano Kokichi, Yamashita Naohide, Nozawa Shiro, Kawakami Yutaka

机构信息

Division of Cellular Signaling, Institute for Advanced Medical Research, Department of Obstetrics and Gynecology, School of Medicine, Keio University, Department of Advanced Medical Science, Institute of Medical Science, University of Tokyo, Tokyo, Japan.

出版信息

Clin Cancer Res. 2005 May 15;11(10):3949-57. doi: 10.1158/1078-0432.CCR-04-1702.

DOI:10.1158/1078-0432.CCR-04-1702

PMID:15897597

Abstract

PURPOSE

Identification of cancer/testis antigens useful for diagnosis or immunotherapy of cancers was attempted by cDNA expression cloning with patients' sera (SEREX).

EXPERIMENTAL DESIGN

cDNA expression libraries made from testis or endometrial cancer cell lines were screened using sera from patients with endometrial cancer or melanoma patients immunized with dendritic cells pulsed with autologous tum or lysates. Tissue-specific expression by RT-PCR and immunogenicity by Western blotting of the bacterial recombinant antigen with sera from cancer patients were evaluated.

RESULTS

A cancer/testis antigen, CAGE, was isolated by two independently performed SEREX. CAGE was expressed in various cancer cell lines including endometrial cancer, colon cancer, and melanoma in 7 of 10 endometrial cancer tissues and in 1 of 3 atypical endometrial hyperplasia, but not in normal tissues including the endometrium and testis. The protein expression on cancer cells was confirmed by Western blot analysis with the recombinant CAGE protein, anti-CAGE IgG antibody was detected in sera from 5 of 45 endometrial cancer, 2 of 24 melanoma, and 2 of 33 colon cancer patients, but not in sera from healthy individuals. By ELISA analysis, anti-CAGE antibody was detected in 12 of 45 endometrial cancer, 2 of 20 melanoma, and 4 of 33 colon cancer patients. Intriguingly, anti-CAGE antibody was highly positive in 7 of the 13 (53.8%) microsatellite instability (MSI)-H patients with endometrial cancer, but negative in 20 non-MSI-H patients (P = 0.001).

CONCLUSION

CAGE may be useful for immunotherapy and diagnosis of various cancers particularly MSI-positive endometrial cancer.

摘要

目的

尝试通过利用患者血清进行cDNA表达克隆（SEREX）来鉴定可用于癌症诊断或免疫治疗的癌/睾丸抗原。

实验设计

使用来自子宫内膜癌患者的血清或用自体肿瘤裂解物脉冲处理的树突状细胞免疫的黑色素瘤患者的血清，筛选由睾丸或子宫内膜癌细胞系构建的cDNA表达文库。通过逆转录聚合酶链反应（RT-PCR）评估细菌重组抗原在组织中的特异性表达，并通过蛋白质印迹法评估其与癌症患者血清的免疫原性。

结果

通过两个独立进行的SEREX分离出一种癌/睾丸抗原CAGE。CAGE在包括子宫内膜癌、结肠癌和黑色素瘤在内的多种癌细胞系中表达，在10个子宫内膜癌组织中的7个、3个非典型子宫内膜增生中的1个中表达，但在包括子宫内膜和睾丸在内的正常组织中不表达。用重组CAGE蛋白进行蛋白质印迹分析证实了癌细胞上的蛋白表达，在45例子宫内膜癌患者中的5例、24例黑色素瘤患者中的2例和33例结肠癌患者中的2例血清中检测到抗CAGE IgG抗体，但在健康个体的血清中未检测到。通过酶联免疫吸附测定（ELISA）分析，在45例子宫内膜癌患者中的12例、20例黑色素瘤患者中的2例和33例结肠癌患者中的4例血清中检测到抗CAGE抗体。有趣的是，在13例（53.8%）微卫星不稳定（MSI）-H的子宫内膜癌患者中的7例中抗CAGE抗体呈高度阳性，但在20例非MSI-H患者中呈阴性（P = 0.001）。