Sialyl-dimeric Lewis-X antigen expressed on mucin-like glycoproteins in colorectal cancer metastases.
Sialyl-dimeric Lewis-X antigen expressed on mucin-like glycoproteins in colorectal cancer metastases.
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发表时间：
1990-12
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通讯作者：
Y. Matsushita;Karen R. Cleary;David M. Ota;S. Hoff;Tatsuro Irimura
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作者：
Y. Matsushita;Karen R. Cleary;David M. Ota;S. Hoff;Tatsuro Irimura
应用特异性单克隆抗体（MAbs）对20例Dukes'C或D期大肠癌原发灶和转移灶进行了N-乙酰乳糖胺寡聚体（NAG）的免疫组化定位和含量测定。所用的单克隆抗体是对刘易斯X抗原特异的SH 1;对二聚体刘易斯X抗原特异的FH 4;对唾液酸二聚体刘易斯X抗原特异的FH 6;和对刘易斯Y-刘易斯X抗原特异的KH 1。这些单克隆抗体鉴定的碳水化合物抗原的分布在原发性肿瘤内以及转移性病变内是异质性的。连续切片的检查表明，用一种单克隆抗体染色的单个肿瘤内的区域并不总是与其他单克隆抗体反应，尽管这四种单克隆抗体鉴定了密切相关的结构。MAb与癌切片的反应程度按阳性癌细胞百分比分类，并比较同一患者的原发肿瘤和转移瘤。转移灶中与MAb FH 6反应的癌细胞比例相等或高于原发性结肠癌，但与其他MAb的相关性未观察到。对肿瘤组织提取物进行电泳分离，然后用这些MAb染色，结果显示分子量约为1，000，000的组分是MAb、FH 6、FH 4和KH 1结合的主要位点。直接单克隆抗体结合所示的聚丙烯酰胺凝胶上的抗原分子的电泳迁移率略有不同的麦胚凝集素在相同的组织中所揭示的一个主要的唾液粘蛋白。通过预先用弱酸或唾液酸酶处理糖蛋白以去除唾液酸，消除了与高分子量组分结合的MAb FH 6。同时，结合单克隆抗体SH 2，特异性二聚体Lex抗原，该组件增加。从肝转移瘤中制备提取物，通过凝胶过滤分离高分子量组分，然后通过DEAE-纤维素离子交换色谱进行分级。从0.10-0.25 M氯化钠之间的DEAE-纤维素洗脱的级分含有大部分MAb FH 6反应性，如抗体亲和层析所示。这些结果支持了一个假设，即大肠癌组织产生的高分子量糖蛋白的糖链是异质性的，其抗原结构可能会在肿瘤进展过程中发生变化。
Colorectal primary carcinomas and metastases from 20 Dukes' stage C or D patients were examined for the immunohistochemical localization and contents of various fucosylated N-acetyl-lactosamine oligomers by specific monoclonal antibodies (MAbs). MAbs used were SH1, specific for Lewis X antigen; FH4, specific for dimeric Lewis X antigen; FH6, specific for sialyl-dimeric Lewis X antigen; and KH1, specific for Lewis Y-Lewis X antigen. The distribution of the carbohydrate antigens identified by these MAbs was heterogeneous within the primary tumor as well as within the metastatic lesion. Examinations of serial sections indicated that areas within an individual tumor which were stained with one MAb were not always reactive with the other MAbs, although these four MAbs identify closely related structures. The degree of MAb reactivity with carcinoma sections was classified by percentage positive carcinoma cells, and primary tumors and metastases from the same patients were compared. An equivalent or higher proportion of carcinoma cells in the metastatic lesions were reactive with MAb FH6 than in the primary colon carcinomas, but each correlation was not seen with the other MAbs. Electrophoretic separation of tumor tissue extracts followed by staining with these MAbs revealed that a component having an approximate molecular weight of 1,000,000 is the major site for the binding of MAbs, FH6, FH4, and KH1. The electrophoretic mobility of the antigenic molecule on polyacrylamide gels as shown by direct MAb bindings was slightly different from that of a major sialomucin revealed by wheat germ agglutinin in the same tissues. MAb FH6 binding to a high molecular weight component was eliminated by prior treatment of the glycoprotein with mild acid or sialidase to remove sialic acid. Simultaneously, binding of MAb SH2, specific for dimeric Lex antigen, to this component increased. An extract was prepared from a liver metastasis, and high molecular weight components were isolated by gel filtration and then fractionated by DEAE-cellulose ion exchange chromatography. A fraction eluted from DEAE-cellulose between 0.10-0.25 M sodium chloride contained most of the MAb FH6 reactivity, as shown by antibody affinity chromatography. These results support a hypothesis that high molecular weight glycoproteins produced by colorectal carcinoma tissues are heterogeneous with regard to their carbohydrate chains and their antigenic structures may change during tumor progression.