Identification of the HERV-K gag antigen in prostate cancer by SEREX using autologous patient serum and its immunogenicity.
Identification of the HERV-K gag antigen in prostate cancer by SEREX using autologous patient serum and its immunogenicity.
复制标题
DOI：
--
复制
发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Toshiaki Ishida;Y. Obata;N. Ohara;H. Matsushita;Shuichiro Sato;A. Uenaka;T. Saika;T. Miyamura;K. Chayama;Yurika Nakamura;H. Wada;T. Yamashita;T. Morishima;L. Old;E. Nakayama
中科院分区：
文献类型：
--
作者：
Toshiaki Ishida;Y. Obata;N. Ohara;H. Matsushita;Shuichiro Sato;A. Uenaka;T. Saika;T. Miyamura;K. Chayama;Yurika Nakamura;H. Wada;T. Yamashita;T. Morishima;L. Old;E. Nakayama
使用自体患者血清通过SEREX分析鉴定前列腺癌HERV-K gag相关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NGO-Pr-54的5 ′端缺失的克隆ZH 042中不断观察到强反应，表明它含有编码蛋白产物的序列。利用ZH 042基因推导的重组蛋白（438 aa）制备TI-35 mAb。将克隆ZH 042转染到293 T细胞中导致产生通过Western印迹可见的约50-kDa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。间接免疫荧光检测显示NGO-Pr-54蛋白在细胞表面和细胞质中均有表达。使用TI-35 mAb通过流式细胞术确认细胞表面表达。在患有膀胱癌（5.1%）、肝癌（4.1%）、肺癌（3.4%）、卵巢癌（5.6%）和前列腺癌（4.2%）以及恶性黑色素瘤（13.2%）的患者中观察到针对NGO-Pr-54的抗体应答。
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%).