Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan ; Department of Urology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan.
Collaborative Research Center for OMIC, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan.
J Immunol Res. 2015;2015:268172. doi: 10.1155/2015/268172. Epub 2015 Mar 25.
Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum. Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN. In this study, we applied the 11-25 mAb to in vivo imaging to detect MSLN-expressing tumors. In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells. We showed the accumulation of Alexa Fluor 750-labeled 11-25 mAb in MSLN-expressing tumor xenografts in athymic nude mice. Then, 11-25 mAb was labeled with (64)Cu via a chelating agent DOTA and was used in both in vitro cell binding assay and in vivo positron emission tomography (PET) imaging in the tumor-bearing mice. We confirmed that (64)Cu-labeled 11-25 mAb highly accumulated in MSLN-expressing tumors as compared to MSLN-negative ones. The (64)Cu-labeled 11-25 mAb is potentially useful as a PET probe capable of being used for wide range of tumors, rather than (18)F-FDG that occasionally provides nonspecific accumulation into the inflammatory lesions.
间皮素(MSLN)是一种 40kDa 的细胞分化相关糖蛋白,出现在癌变过程中,在许多人类癌症中高度表达,包括大多数胰腺腺癌、卵巢癌和间皮瘤,而在正常组织中的表达仅限于覆盖胸膜、心包和腹膜的间皮细胞。克隆 11-25 是一种分泌针对人 MSLN 的单克隆抗体(mAb)的鼠杂交瘤。在这项研究中,我们应用 11-25 mAb 进行体内成像以检测表达 MSLN 的肿瘤。在体外和离体免疫化学研究中,我们证明了 11-25 mAb 对几种胰腺癌细胞表面膜结合 MSLN 的特异性。我们表明,Alexa Fluor 750 标记的 11-25 mAb 在无胸腺裸鼠的表达 MSLN 的肿瘤异种移植中积累。然后,通过螯合剂 DOTA 将 11-25 mAb 标记上(64)Cu,并在携带肿瘤的小鼠中进行体外细胞结合测定和体内正电子发射断层扫描(PET)成像。我们证实,与 MSLN 阴性肿瘤相比,(64)Cu 标记的 11-25 mAb 在表达 MSLN 的肿瘤中高度积累。(64)Cu 标记的 11-25 mAb 有望作为一种 PET 探针,可用于广泛的肿瘤,而不是(18)F-FDG,后者偶尔会导致炎症病变的非特异性积累。