CHARACTERIZATION OF HUMAN CARCINOMA T & TN AUTOANTIGENS

人类癌症的特征

• 批准号: 3165842
• 负责人: GEORG F SPRINGER
• 金额: $ 13.27万
• 依托单位: ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-06-01 至 1993-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3165842
• 关键词: adenocarcinoma; affinity chromatography; autoantigens; blood group antigens; breast neoplasms; carbohydrate structure; cellular oncology; gas chromatography mass spectrometry; glycolipids; glycopeptides; glycoprotein structure; glycoproteins; guinea pigs; human tissue; immunochemistry; immunoperoxidase; metastasis; molecular oncology; monoclonal antibody; neoplasm /cancer immunology; neoplastic cell; nuclear magnetic resonance spectroscopy; preneoplastic state; protein sequence; protein structure; tissue /cell culture; tumor antigens

Immunoreactive Thomsen-Friedenreich (T) antigen and its precursor, Tn, occur in approximately 90% of all human (adeno) carcinomas (CAs) and in their metastases. In nonCA tissues, these antigens are occluded and inaccessible to the immune system. T & Tn are the immediate precursors of blood type M,N antigens. T & Tn are the first universal CA-diagnostic markers; they are adhesion molecules and persistent autoantigens, from incipience onward; T & Tn's surface densities are often prognostic. Our proposed structures of the T & Tn epitopes based on stepwise degradation and biosynthesis were later confirmed by organic synthesis. The focus will be on Stage I ductal breast CAs. We plan: Study of fresh surgical specimens of primary, preferably Stage I, CAs and tissue-cultured CAs plus control tissues. Full characterization of CA-T & -Tn's structure, location and integration in carrier molecules, macromolecular conformation and distribution in CA. To define extent of T & Tn epitope-clustering, and to determine I & Tn epitopes' interrelationship in CA using metabolically labeled breast CA cells. Immunohistochemical evaluation (peroxidase, gold- silver) with light (ultra)microscopy of spatial distribution of T & Tn epitopes with our anti-T & -Tn rodent monoclonal (Mo) and human monospecific, polyclonal antibodies (Abs). Quantitation of T & Tn epitopes on breast CA cells and on control tissues by RIA and by computerized image analysis, relating T & Tn densities of CA to their ploidy, and correlation of T & Tn densities to invasiveness and autoimmunogenicity of different breast CAs. Lectin and Ab affinity chromatography (C) to separate CA glycoproteins based on their predominant Tn, T and N & M epitopes. Purification of T- & Tn-active glycopeptides obtained by pronase digestion and their reductively beta-eliminated oligosaccharides (OS) by gel filtration, ion-exchange & high-performance liquid C. To determine OS composition by hydrolysis, enzymatically, colorimetrically and by gas C (GC) of alditol acetates, and structures using glycosidases, permethylation, GC-mass spectrometry (GC/MS) & NMR. Amino acid sequencing will be by subtractive Edman degradation. To isolate guinea pig L-lO hepatoCA Tn-, T- and N- & M-active glycolipids (GLs) on s large scale, and define the carbohydrate structures of the T- & Tn-specific GLs. Study of human CAs and lymphomas for expression and nature of T- & Tn-active GLs; relate findings to degree of malignancy. Immunostaining with our MoAbs to locate and identify active GLs. To analyze OS of active L-lO GLs by GC/MS after trifluoroacetolysis, reduction and permethylation.

免疫反应性Thomsen-Friedenreich（T）抗原及其前体， TN，大约90％的人（Adeno）癌症发生 （CAS）及其转移。 在非CA组织中，这些抗原 免疫系统被阻塞和无法访问。 T＆TN是 血型M，N抗原的直接前体。 T＆TN是 第一个通用CA诊断标记；它们是粘附分子 从开始时，持续的自动抗原； T＆TN的 表面密度通常是预后的。我们提出的结构 基于逐步降解和生物合成的T＆TN表位 后来通过有机合成证实。 重点将开始 阶段I导管乳房。 我们计划：新的手术研究 主要的，优选的I期，CA和组织培养的标本 CAS加控制组织。 CA -T＆-TN的完整表征 载体分子中的结构，位置和整合， 大分子构象和分布 定义 T＆TN表位聚类的范围，并确定I＆TN 使用代谢标记的CA中的表位相互关系 乳房Ca细胞。 免疫组织化学评估（过氧化物酶，金 - 银色），带有t的光（超）显微镜 ＆TN表位与我们的抗T＆-TN啮齿动物单克隆（MO）和 人单特异性，多克隆抗体（ABS）。 定量 乳房Ca细胞和对照组织上的T＆TN表位RIA 通过计算机图像分析，将T＆TN密度相关 CA与它们的拼合物以及T＆TN密度的相关性 不同乳腺Cas的侵入性和自身免疫原性。 凝集素和AB亲和色谱（C）分离Ca 基于其主要TN，T和N＆M表位的糖蛋白。 通过prenase获得的T-和TN活性糖肽的纯化 消化及其还原的β-降低寡糖 （OS）通过凝胶过滤，离子 - 交换和高性能液体C。 通过酶促的水解确定OS组成， 比色法和乙酸醛醇的气C（GC），以及 使用糖苷酶，苄丙二酰化，GC质量光谱法的结构 （GC/MS）和NMR。氨基酸测序将通过减法Edman 降解。分离豚鼠l-lo Hepatoca tn-，t-和n-＆＆ 大规模上的M活性糖脂（GLS），并定义 T和TN特异性GL的碳水化合物结构。 研究 人CA和淋巴瘤，用于T-和TN活性的表达和性质 GLS；将发现与恶性肿瘤程度有关。 免疫染色 我们的摩押可定位和识别活跃的GL。 分析OS 三氟乙酸酯，还原和降低和 苄苄氨基化。

项目成果

Highly active amino-terminal sialoglycopenta-, hexa- and heptapeptides from human erythrocyte NM antigens.

来自人红细胞 NM 抗原的高活性氨基末端唾液酸糖五肽、六肽和七肽。

• DOI: 10.1007/bf00405655
• 发表时间: 1980
• 期刊: Die Naturwissenschaften
• 作者: Springer,GF;Yang,HJ;Mbawa,E;Grohlich,D;Friedenson,B
• 通讯作者: Friedenson,B

Detection of lung- and breast carcinoma by quantitating serum anti-T IgM levels with a sensitive, solid-phase immunoassay.

通过灵敏的固相免疫分析定量血清抗 T IgM 水平来检测肺癌和乳腺癌。

• DOI: 10.1007/bf00480463
• 发表时间: 1982
• 期刊: Die Naturwissenschaften
• 作者: Springer,GF;Desai,PR
• 通讯作者: Desai,PR

Blood group T and Tn antigens are universal, clonal, epithelial cell-adhesive, autoimmunogenic carcinoma markers.

T 血型和 Tn 抗原是通用的、克隆的、上皮细胞粘附的、自身免疫性癌标记物。

• 发表时间: 1983
• 期刊: Progress in clinical and biological research
• 作者: Springer,GF

Cross-reaction of anti-asialoganglioside sera with human blood group N antigen.

抗去唾液酸神经节苷脂血清与人血型 N 抗原的交叉反应。

• DOI: 10.1007/bf01047265
• 发表时间: 1981
• 期刊: Die Naturwissenschaften
• 作者: Springer,GF;Tegtmeyer,H
• 通讯作者: Tegtmeyer,H

Concurrent immunohistochemical staining of tumor-infiltrating lymphocytes and carcinoma-associated T (Thomsen-Friedenreich)/Tn antigens in human breast carcinoma.

对人乳腺癌中肿瘤浸润淋巴细胞和癌相关 T (Thomsen-Friedenreich)/Tn 抗原进行同步免疫组织化学染色。

• DOI: 10.1177/44.2.8609376
• 发表时间: 1996
• 期刊: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
• 作者: Wang,BL;Springer,GF;Kaufman,MW
• 通讯作者: Kaufman,MW