Ishida Toshiaki, Obata Yuichi, Ohara Nobuya, Matsushita Hirokazu, Sato Shuichiro, Uenaka Akiko, Saika Takashi, Miyamura Takako, Chayama Kosuke, Nakamura Yurika, Wada Hisashi, Yamashita Toshiharu, Morishima Tsuneo, Old Lloyd J, Nakayama Eiichi
Department of Immunology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
Cancer Immun. 2008 Nov 13;8:15.
The prostate cancer HERV-K gag-related NGO-Pr-54 antigen was identified by SEREX analysis using autologous patient serum. NGO-Pr-54 mRNA was observed to be faintly expressed in normal prostate and strongly expressed in a variety of cancers, including ovarian cancer (5/8), prostate cancer (6/9), and leukemia (5/14). A phage plaque assay showed that a strong reaction was constantly observed with clone ZH042 in which the 5' end of NGO-Pr-54 is deleted, suggesting that it contained the sequence coding for the protein product. A TI-35 mAb was produced using a recombinant protein (438 aa) deduced from the sequence of ZH042. Transfection of clone ZH042 into 293T cells resulted in the production of an approximately 50-kDa molecule visualized by Western blotting. Natural production of the molecule was confirmed in a SK-MEL-23 melanoma cell line. An indirect immunofluorescence assay showed that NGO-Pr-54 protein was expressed on the cell surface as well as in the cytoplasm. Cell surface expression was confirmed by flow cytometry using the TI-35 mAb. The antibody response against NGO-Pr-54 was observed in patients with bladder (5.1%), liver (4.1%), lung (3.4%), ovarian (5.6%), and prostate (4.2%) cancer, as well as with malignant melanoma (13.2%).
使用患者自身血清通过SEREX分析鉴定出前列腺癌人内源性逆转录病毒-K gag相关的NGO-Pr-54抗原。观察到NGO-Pr-54 mRNA在正常前列腺中微弱表达，而在多种癌症中强烈表达，包括卵巢癌（5/8）、前列腺癌（6/9）和白血病（5/14）。噬菌体噬斑试验表明，在缺失NGO-Pr-54 5'端的克隆ZH042中持续观察到强烈反应，这表明它包含编码该蛋白质产物的序列。使用从ZH042序列推导的重组蛋白（438个氨基酸）制备了TI-35单克隆抗体。将克隆ZH042转染到293T细胞中，通过蛋白质印迹法可观察到产生了一个约50 kDa的分子。在SK-MEL-23黑色素瘤细胞系中证实了该分子的天然产生。间接免疫荧光试验表明，NGO-Pr-54蛋白在细胞表面和细胞质中均有表达。使用TI-35单克隆抗体通过流式细胞术证实了细胞表面表达。在膀胱癌（5.1%）、肝癌（4.1%）、肺癌（3.4%）、卵巢癌（5.6%）、前列腺癌（4.2%）患者以及恶性黑色素瘤患者（13.2%）中观察到了针对NGO-Pr-54的抗体反应。
