Dube V E, Kallio P, Chmiel J S, Haid M, Hakim A
Department of Pathology and Laboratory Medicine, Evanston Hospital, Illinois 60201.
Clin Immunol Immunopathol. 1987 Nov;45(2):196-207. doi: 10.1016/0090-1229(87)90034-1.
Tumor cells elaborate and release into the circulation a variety of glycoproteins. An enzyme-linked immunosorbent assay (ELISA) was developed to monitor carbohydrate structures secreted into the circulation. Among these antigens are the structures specific for the blood group I antigens, which are incompletely converted to ABH antigens on the membranes of tumor cells. The I antigens in the sera of 67 women with breast carcinoma (BCa), 58 with benign breast disease (BBD), and 47 controls were measured by the ELISA. In this assay, I antigen from ovarian cyst mucin was bound to the wells of polystyrene microtiter plates. The monoclonal human anti-I antibody (Hy) was added to the wells along with perchloric acid extracts of patient and control sera at five different dilutions. The anti-I binding to the solid-phase I antigen was determined after incubation steps with peroxidase-labeled anti-human IgM and substrate. The amount of sera extracts giving 50% inhibition of anti-I (Hy) binding was determined from the inhibition curves which were corrected by integrating the slope values into that of the standard curve obtained with extracts of normal sera. The I antigens were significantly higher in pathologic stage (PS) IV sera (P less than 0.001), and comparable in PS I, PS II, and PS III and BBD sera to those in control sera. The anti-I (Hy) binds strongly Gal 1,4 GlcNAc 1,6 Gal (alpha GalNAc); Gal 1,4 GlcNAc 1,6 (Gal 1,4 GlcNAc 1,3) Gal; and to a lesser extent Gal 1,4 GlcNAc 1,3 Gal 1,4 GlcNAc (0.06, 0.09, and 0.35 mM, giving 50% inhibition, respectively). It was concluded that similar or related structures may be expressed on the membrane of metastatic BCa cells.
肿瘤细胞合成并释放多种糖蛋白进入循环系统。开发了一种酶联免疫吸附测定（ELISA）来监测分泌到循环系统中的碳水化合物结构。这些抗原中包括血型I抗原的特异性结构，它们在肿瘤细胞膜上未完全转化为ABH抗原。通过ELISA检测了67例乳腺癌（BCa）女性、58例良性乳腺疾病（BBD）女性和47例对照者血清中的I抗原。在该测定中，将来自卵巢囊肿粘蛋白的I抗原结合到聚苯乙烯微量滴定板的孔中。将单克隆人抗I抗体（Hy）与患者和对照血清的高氯酸提取物以五种不同稀释度一起加入孔中。在用过氧化物酶标记的抗人IgM和底物进行孵育步骤后，测定抗I与固相I抗原的结合。从抑制曲线确定给予抗I（Hy）结合50%抑制的血清提取物量，该抑制曲线通过将斜率值整合到用正常血清提取物获得的标准曲线斜率值中进行校正。I抗原在病理分期（PS）IV血清中显著更高（P小于0.001），在PS I、PS II和PS III以及BBD血清中与对照血清中的相当。抗I（Hy）与Gal 1,4 GlcNAc 1,6 Gal（αGalNAc）；Gal 1,4 GlcNAc 1,6（Gal 1,4 GlcNAc 1,3）Gal强烈结合；并在较小程度上与Gal 1,4 GlcNAc 1,3 Gal 1,4 GlcNAc结合（分别为0.06、0.09和0.35 mM，产生50%抑制）。得出的结论是，转移性BCa细胞膜上可能表达相似或相关的结构。
