Transduction and expression of the human carcinoembryonic antigen gene in a murine colon carcinoma cell line.
Transduction and expression of the human carcinoembryonic antigen gene in a murine colon carcinoma cell line.
复制标题
人癌胚抗原基因在鼠结肠癌细胞系中的转导和表达。
作者：
P. Robbins;Judy Kantor;M. Salgaller;P. Hand;Philip D. Fernsten;Jeffrey Schlom
源自小鼠结肠腺癌 MC-38 的细胞系已用含有编码人癌胚抗原 (CEA) 基因的互补 DNA 的逆转录病毒构建体转导。 MC-38 在同基因 C57BL/6 小鼠中形成肿瘤，作为主动免疫疗法的靶点已被广泛研究。分离了表达高水平细胞表面 CEA 的单个转导克隆，并对两个克隆（称为 MC-38-ceal 和 MC-38-cea2）进行了广泛表征。这些克隆表面的 CEA 水平显着高于中等分化的人结肠癌细胞系 (WiDr) 中的水平，并且与人结肠癌细胞系 GEO 和 CBS（报道的 CEA 表达最高的细胞之一）中的水平相当。进一步分析表明，MC-38-cea1 克隆中表达的 CEA 具有与天然 CEA (Mr 180,000) 相似的分子量，但 MC-38-cea2 细胞系表达单个 Mr 70,000 糖基化免疫反应产物。发现七种抗 CEA 单克隆抗体与两个克隆发生反应。对 MC-38-cea2 克隆中存在的 CEA 基因进行了部分测序，发现 CEA 中存在的三个重复结构域中的两个缺失。这些结果为未来的研究奠定了基础，以绘制 CEA 的免疫显性表位，并开发同基因模型系统，该系统可能有助于设计试剂和方案，以研究针对表达人类 CEA 的癌症的主动和被动免疫疗法。
A cell line derived from the mouse colon adenocarcinoma, MC-38, has been transduced with a retroviral construct containing complementary DNA encoding the human carcinoembryonic antigen (CEA) gene. MC-38, which forms tumors in syngeneic C57BL/6 mice, has been extensively studied as a target for active immunotherapy. Individual transduced clones that express high levels of cell surface CEA were isolated, and two clones, termed MC-38-ceal and MC-38-cea2, were extensively characterized. The levels of CEA found on the surface of these clones were considerably higher than that found in a moderately differentiated human colon carcinoma cell line (WiDr) and were comparable to those found on the human colon carcinoma cell lines GEO and CBS (among the highest CEA-expressing cells reported). Further analysis demonstrated that the CEA expressed in the MC-38-cea1 clone had a similar molecular weight to native CEA (Mr 180,000), but the MC-38-cea2 cell line expressed a single Mr 70,000 glycosylated immunoreactive product. Seven anti-CEA monoclonal antibodies were found to react with both clones. The CEA gene present in the MC-38-cea2 clone was partially sequenced and was found to contain a deletion of two of the three repeated domains present in CEA. These results provide a basis for future studies to map immunodominant epitopes of CEA and to develop a syngeneic model system that may aid in the design of reagents and protocols to study active and passive immunotherapy directed against a carcinoma expressing human CEA.
登录
查看更多内容
DOI：
10.1073/pnas.85.13.4648
发表时间：
1988-07-01
影响因子：
11.1
作者：
HEFTA, SA;HEFTA, LJF;SHIVELY, JE
通讯作者：
SHIVELY, JE
单克隆抗体 KS1/4 识别的癌相关抗原的生物合成和糖基化。
影响因子：
11.2
作者：
Fernsten,PD;Pekny,KW;Reisfeld,RA;Walker,LE
通讯作者：
Walker,LE
双决定簇放射免疫分析中使用单克隆抗体检测胃肠道肿瘤患者血清中的癌胚抗原和相关抗原。
影响因子：
--
作者：
Herlyn,M;Blaszczyk,M;Sears,HF;Verrill,H;Lindgren,J;Colcher,D;Steplewski,Z;Schlom,J;Koprowski,H
通讯作者：
Koprowski,H
癌胚抗原和非特异性交叉反应抗原的互补DNA和基因组克隆在中国仓鼠卵巢和小鼠成纤维细胞中的表达以及膜表达产物的表征。
影响因子：
11.2
作者：
Hefta,LJ;Schrewe,H;Thompson,JA;Oikawa,S;Nakazato,H;Shively,JE
通讯作者：
Shively,JE
逆转录病毒载体转导的兔内皮细胞中高水平重组基因表达。
DOI：
10.1126/science.2911735
发表时间：
1989
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Zwiebel,JA;Freeman,SM;Kantoff,PW;Cornetta,K;Ryan,US;Anderson,WF
通讯作者：
Anderson,WF