Sata T, Roth J, Zuber C, Stamm B, Rinderle S J, Goldstein I J, Heitz P U
Department of Pathology, National Institute of Health, Tokyo, Japan.
Lab Invest. 1992 Feb;66(2):175-86.
The lectin Amaranthin has been shown to be highly specific for the galactose beta 1,3 N-acetylgalactosamine-alpha and sialic acid alpha 2,3 galactose beta 1,3 N-acetylgalactosamine-alpha sequence which represents the Thomsen-Friedenreich (T) antigen and its cryptic form, respectively. Previously, we demonstrated the usefulness of gold-labeled Amaranthin for the histochemical detection of the T antigen and its cryptic form. Application of the galactose oxidase (GO)-Schiff sequence abolished lectin binding to the T antigen but not its cryptic form, and therefore permitted their differentiation. In the present study we have analyzed by light and electron microscopy the distribution and subcellular localization of Amaranthin binding sites in normal, dysplastic and neoplastic colonic epithelium. Furthermore, a monoclonal antibody raised against synthetic galactose bera 1,3 N-acetylgalactosamine-alpha-bovine serum albumin was applied as a reagent for the T antigen. In normal colonic mucosa, two different Amaranthin staining patterns existed: (a) reactivity restricted to the lower portion of the crypts which was principally observed in the left colon, and (b) reactivity along the entire length of the crypts and in the surface epithelium with goblet cell staining in the upper portion of the crypts which was principally observed in the right colon. This Amaranthin staining was resistant to GO-Schiff treatment. No immunostaining with the monoclonal anti-T antigen was observed. Investigation of transitional mucosa, adenocarcinomas of different degrees of differentiation and mucinous carcinomas as well as adenomas with different degrees of dysplasia all revealed positive Amaranthin staining. The lectin staining was resistant to GO-Schiff treatment, and immunolabeling with the monoclonal antibody against the T antigen was absent. These results indicate that only the cryptic form of the T antigen is expressed in normal, dysplastic and neoplastic human colonic epithelium.
凝集素苋菜红已被证明对半乳糖β1,3 N-乙酰半乳糖胺-α和唾液酸α2,3半乳糖β1,3 N-乙酰半乳糖胺-α序列具有高度特异性，这两种序列分别代表了汤姆森-弗里德赖希（T）抗原及其隐蔽形式。此前，我们证明了金标记的苋菜红在组织化学检测T抗原及其隐蔽形式方面的实用性。半乳糖氧化酶（GO）-席夫序列的应用消除了凝集素与T抗原的结合，但未消除其与隐蔽形式的结合，因此可以区分它们。在本研究中，我们通过光学显微镜和电子显微镜分析了苋菜红结合位点在正常、发育异常和肿瘤性结肠上皮中的分布和亚细胞定位。此外，一种针对合成半乳糖β1,3 N-乙酰半乳糖胺-α-牛血清白蛋白产生的单克隆抗体被用作T抗原的试剂。在正常结肠黏膜中，存在两种不同的苋菜红染色模式：（a）反应性局限于隐窝下部，主要在左结肠观察到；（b）反应性沿隐窝全长以及表面上皮存在，隐窝上部杯状细胞染色，主要在右结肠观察到。这种苋菜红染色对GO-席夫处理具有抗性。未观察到单克隆抗T抗原的免疫染色。对移行黏膜、不同分化程度的腺癌和黏液腺癌以及不同发育异常程度的腺瘤的研究均显示苋菜红染色呈阳性。凝集素染色对GO-席夫处理具有抗性，且不存在针对T抗原的单克隆抗体的免疫标记。这些结果表明，只有T抗原的隐蔽形式在正常、发育异常和肿瘤性人类结肠上皮中表达。
