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（CA）相关的TN抗原 除了T。我们发现TN是T的直接前体。 通用的人类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经常预后的情况下有力 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 对体液抗T反应很重要。 在体外模型中，我们显示了 t- \和TN特异性簇非常参与Ca粘附 壁纸组织，这是CA入侵的主要步骤之一。 使用 les缩的人类O红细胞（T RBC）我们制备了大鼠杂交瘤 产生单克隆抗T，另外产生单克隆的抗T 抗TN抗体与末端反应 n-乙酰基-D-摩乳氨基氨基烷酰基-O-（galnac-alpha-o-）结构。 随后，我们证明了完全，特别是脱透了的O rbc和t 抗原从中分离出来，具有表达Tn的Galnac-Alpha-o- \分组 用普通人多隆抗TN确定的特异性 抗体。 这一发现似乎需要修改 一般观点，罕见的天然发生的TN是A的结果 “ 3-d-d-d-galactosylsylansferase酶的选择性缺陷。” 而是 导致TN的体细胞突变可能具有多效效应 涉及辅助酶和半乳糖基转移酶。 T rbc衍生的T抗原 （伪影）因此类似于自然细胞表面结构 分化良好的CAS，我们最近显示出比TN多的T； 对于塑料CAS，相反的情况通常是正确的。 14个单克隆中的四个 其他其他人给出的抗人Ca抗体被证明是抗TN。这 表示TN在CA中的进口。 与一般信念相反，我们检测到 人类胚胎中的免疫反应性T和TN表位（6至12周龄） 体现阶段）。 关于健康人RBC的生化研究，几内亚 猪L-10肝癌，在人类乳腺CA培养上（4 x 10 11个细胞具有 现在种植）导致糖肽（GP）和糖脂（GL）分数 与T，TN，N和M活性。 从加拿大皇家银行，我们获得了同质的 主动NH 2-末端五角杆，六角形和Hepta-GPS（约2 kDa） 具有T特异性簇和较大的TN和TN主动GPS（约12个 KDA）。 从L-10 CA细胞中，我们获得了T和TN主动中性 大于四亚己糖基神经酰胺和N和M活性的糖脂 神经节;用A. ureafaciens唾液酸酶治疗更大的 其N和M活动的75％，并引起了共生醇阴性 分量，色谱与ASIALO-G M1相似，具有T活性。我们 在膜和细胞质中鉴定的T- \和TN特异性糖蛋白 培养的人类乳腺CA细胞的分数并部分纯化这些 通过DEAE-纤维素分馏和凝胶过滤的抗原在Sephacryl上进行过滤 S-200。 T活动材料具有GAL和GALNAC（摩尔比大约 1.5：1）。 TN活性GP馏分富含终端Galnac，Ser和Ser和 THR在两者中都是突出的。 （AG）