NATURE OF T-SPECIFIC HUMAN CARCINOMA ANTIGENS

T 特异性人类癌抗原的性质

• 批准号: 3165840
• 负责人: GEORG F SPRINGER
• 金额: $ 12.95万
• 依托单位: EVANSTON HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-08-01 至 1987-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3165840
• 关键词: adenocarcinoma; affinity chromatography; blood group antigens; breast neoplasms; chemical structure; erythrocytes; galactosyltransferases; glycolipids; glycopeptides; human tissue; immunochemistry; liver neoplasms; metastasis; molecular oncology; neoplasm /cancer immunology; neoplastic cell; radiotracer; sialate; tissue /cell culture; tumor antigens

Progress in 1984 to 1985 includes carcinoma (CA)-associated Tn antigen in addition to T. We found Tn to be the immediate precursor of T. Both are universal human CA markers and autoantigens. In the former role, they are powerful in pathohistological diagnosis and frequently prognosis, while cellular and humoral anti-T autoimmune responses permit early CA detection with high sensitivity and specificity. Conformational factors are important for humoral anti-T responses. In an in vitro model, we showed T-\and Tn-specific clusters to be strongly involved in CA adhesion to hearthy tissue, one of the primary steps of CA invasion. Using desialylated human O red blood cells (T RBC) we prepared rat hybridomas producing monoclonal anti-T, and in addition those producing monoclonal anti-Tn antibodies reacting with terminal N-acetyl-d-galactosaminoyl-alpha-O-(GalNAc-alpha-O-) structures. Subsequently we showed that fully, specifically desialylated O RBC, and T antigen isolated therefrom, possess GalNAc-alpha-O-\groupings expressing Tn specificity as determined with ordinary human polychlonal anti-Tn antibodies. This finding would appear to necessitate revision of the general view that the rare naturally occurring Tn is the result of a "selective deficiency of 3-beta-d-galactosyltransferase." Rather, the somatic mutation leading to Tn is likely to have a pleiotropic effect involving both sialyl-\and galactosyltransferases. T RBC-derived T antigen (an artifact) thus resembles natural cell surface structures of well-differentiated CAs, which we recently showed to have more T than Tn; the reverse was generally true for anaplastic CAs. Four of 14 monoclonal anti-human CA antibodies given by others turned out to be anti-Tn; this indicates Tn's import in CA. Contrary to general belief, we detected immunoreactive T and Tn epitopes in human embryos (6 to 12-week-old, pretolerant stage). Biochemical studies on healthy human RBC, on guinea pig L-10 hepatoCA, and on a human breast CA culture (4 x 1011 cells have now been grown) resulted in glycopeptide (GP) and glycolipid (GL) fractions with T, Tn, N, and M activity. From the RBCs we obtained homogeneous active NH2-terminal penta-, hexa-, and hepta-GPs (approximately 2 kDa) with T-specific clusters and larger, T and Tn active GPs (approximately 12 kDa). From the L-10 CA cells, we obtained T and Tn active neutral glycolipids larger than tetrahexaosyl ceramide and N and M active gangliosides; treatment with A. ureafaciens sialidase destroyed greater than 75% of their N and M activities and gave rise to a resorcinol negative component, chromatographically similar to asialo-GM1, with T activity. We identified T-\and Tn-specific glycoproteins in membrane and cytoplasmic fractions of cultured human breast CA cells and partially purified these antigens by DEAE-cellulose fractionation and gel filtration on Sephacryl S-200. T active material has Gal and GalNAc (molar ratio approximately 1.5:1). The Tn active GP fraction is rich in terminal GalNAc, and Ser and Thr are prominent in both. (AG)

1984 年至 1985 年的进展包括癌症 (CA) 相关 Tn 抗原 除了 T 之外。我们发现 Tn 是 T 的直接前身。两者都是 通用人类 CA 标记和自身抗原。 在前一个角色中，他们是 在病理组织学诊断和预后方面具有强大的作用，同时 细胞和体液抗 T 自身免疫反应允许早期 CA 检测 具有高灵敏度和特异性。 构象因素是 对于体液抗 T 反应很重要。 在体外模型中，我们展示了 T-\和 Tn 特异性簇强烈参与 CA 粘附 健康组织，CA 侵袭的主要步骤之一。 使用 我们制备了大鼠杂交瘤去唾液酸化人O红细胞（T RBC） 生产单克隆抗-T，此外还生产单克隆 抗 Tn 抗体与末端反应 N-乙酰基-d-半乳糖氨基酰基-α-O-(GalNAc-α-O-)结构。 随后我们证明了完全、特异性脱唾液酸化的 O RB​​C 和 T 从其分离的抗原，具有表达 Tn 的 GalNAc-α-O-\groupings 用普通人多克隆抗 Tn 测定的特异性 抗体。 这一发现似乎需要修订 一般认为，罕见的自然发生的 Tn 是 “3-β-d-半乳糖基转移酶的选择性缺陷。” 相反， 导致 Tn 的体细胞突变可能具有多效性效应 涉及唾液酸转移酶和半乳糖基转移酶。 T RBC衍生的T抗原 （人工制品）因此类似于自然细胞表面结构 分化良好的 CA，我们最近证明其 T 多于 Tn； 对于间变性 CA 来说，情况通常正好相反。 14 个单克隆中的四个 别人给予的抗人CA抗体结果是抗Tn；这 表示 Tn 在 CA 中的导入。 与普遍看法相反，我们发现 人类胚胎（6至12周龄， 耐受前阶段）。 在几内亚对健康人红细胞进行生化研究 猪 L-10 hepatoCA，以及人乳腺 CA 培养物（4 x 10 11 细胞具有 现已生长）产生糖肽（GP）和糖脂（GL）级分 具有 T、Tn、N 和 M 活性。 从红细胞中我们获得了均质的 活性 NH 2 -末端五-、六-和七-GP（约 2 kDa） 具有 T 特异性簇和更大的 T 和 Tn 活性 GP（大约 12 kDa）。 从 L-10 CA 细胞中，我们获得了 T 和 Tn 活性中性 糖脂大于四己糖基神经酰胺且 N 和 M 具有活性 神经节苷脂；用A. ureafaciens唾液酸酶处理破坏更大 超过 75% 的 N 和 M 活性，并产生间苯二酚阴性 组分，色谱上与 asialo-G M1 类似，具有 T 活性。我们 鉴定出膜和细胞质中的 T-\ 和 Tn 特异性糖蛋白 培养的人乳腺 CA 细胞的部分并部分纯化这些 通过DEAE-纤维素分级分离和Sephacryl凝胶过滤来分离抗原 S-200。 T活性物质有Gal和GalNAc（摩尔比约为 1.5:1）。 Tn 活性 GP 部分富含末端 GalNAc、Ser 和 Thr 在两者中都很突出。 (AG)