Identification of new cervical cancer antigens by SEREX using sera from cervical cancer tissue transplanted SCID mice

在这项研究中，我们利用各种新的免疫学方法，鉴定了一种新的癌症抗原，它不仅是一种有用的免疫治疗靶点，而且是诊断子宫腺癌的生物标志物。我们采集了19例宫颈癌手术患者的肿瘤组织样本和29例患者的血液样本。我们通过HPV- e7特异性PCR和测序确定了这些组织中HPV感染的存在和HPV类型。在8例鳞状细胞癌中，检出HPV 16相关型4例（16例、31例、58例），HPV 18相关型1例（39例）。在9例腺癌中，检出HPV 18型4例，HPV 16型2例。从13例宫颈癌样本中，我们将癌组织移植到SCID小鼠体内。在这些SCID小鼠的肿瘤浸润性B淋巴细胞中检测人IgG后，我们收集了血清。我们从宫颈癌细胞系SKG-II/SKG-IIIb、TCO-1、Hela构建了lambda噬菌体cDNA表达文库。此外，我们从4例癌组织中构建了更多的lambda噬菌体cDNA表达文库。首先，我们用肿瘤移植SCID小鼠血清筛选TOC-1文库。在HPV-18阳性腺癌的病例中，NY-REN-58被报道为肾癌的候选癌症抗原，使用这种方法从这些血清中鉴定出来。肿瘤排斥抗原-1 （Tumor rejection antigen-1, gp96）是抗原提呈细胞的分子伴侣，因其与肿瘤免疫相关而被研究用于肽疫苗治疗。然后，将转移组织与HPV 18阳性宫颈腺癌的原发组织分开移植，用原发和转移两种血清筛选SKG-II/IIIb文库。我们分离了50万个cDNA克隆，但没有确定一个阳性克隆。然后我们利用每次筛选的睾丸文库分离出400多个阳性克隆，目前我们正在继续对这些阳性克隆进行调查。我们将利用植瘤SCID小鼠血清筛选HPV 18阳性癌组织或HPV 16阳性癌组织的其他文库，确定宫颈癌抗原用于临床诊断和治疗。少

In this study, using new various immunological methods, we identified a novel cancer-antigen which is useful not only for an immune therapy target but also a biomarker for the diagnosis of adenocarcinoma in the uterine cervix. We obtained 19 tumor tissue samples and 29 blood samples from cervical cancer patients at operation. We determined the presence of HPV infection of these tissues and the HPV type by HPV-E7 specific PCR and sequencing. In squamous cell carcinoma 8 cases, we detected 4 cases of HPV 16-related type (16,31,58) and 1 case of HPV 18-related type (39). In 9 adenocarcinoma cases, we detected 4 cases of HPV 18-type and 2 cases of HPV 16-type. From 13 cervical cancer samples, we transplanted the cancer tissue into SCID mice. After detecting human IgG in these SCID mice from tumor-infiltrating B lymphocytes, we harvested the sera. We constructed lambda phage cDNA expression libraries from cervical cancer cell lines, SKG-II/SKG-IIIb, TCO-1, Hela. In addition, we constructed … More lambda phage cDNA expression libraries from 4 cases of cancer tissues. First, we screened TOC-1 library with the sera from tumor transplanted SCID mice. In the case of HPV-18 positive adenocarcinoma, NY-REN-58, reported as candidate of cancer antigen in renal cancer, was identified from these sera using this method. Tumor rejection antigen-1 (gp96) was reported to be a molecular chaperon of antigen presenting cells, and studied for peptide vaccine therapy application because of its association with tumor immunity. Then, transplanted the metastatic tissue separately from primary tissue of HPV 18 positive cervical adenocarcinoma, we screened SKG-II/IIIb library with two sera from primary and metastatic. We isolated 500,000 cDNA clones, but a positive clone was not determined. Then we isolated over 400 positive clones using a testis library from each screening, and we are currently continuing the investigation of these positive clones. We will screen other libraries from HPV 18-positive cancer tissue or HPV 16-positive cancer tissue with the sera of tumor-planted SCID mice and identify a cancer antigen for clinical diagnosis and treatment of cervical adenocarcinoma. Less