BREAST EPITHELIAL ANTIGENS AS TARGETS FOR RIA

参加此次活动的两个项目的总体目标是发展 并测试新的基于单抗的乳房治疗策略 癌症。该项目将探索乳房的分子和细胞生物学。 最好的上皮抗原(BEAs)及其表位结构 放射免疫治疗(RLT)的乳腺肿瘤定位靶点。 项目2旨在识别乳房识别的人类B淋巴细胞 肿瘤在其人类宿主中，产生由 通过建立稳定的细胞系来产生这些细胞 基因工程，并开发放射免疫治疗的新策略 通过人性化和碎片化现有的MoAbs翻译成 临床试验。 该项目的具体目标是： 1.分离和鉴定乳房粘蛋白的细胞相关形式和 BA46，并使用 我们已经开发出的移动抗体。 2.人MoAbs鉴定的BEA的分离和鉴定 在项目2中开发。区分细胞相关形式和分泌形式。 3.分离并测序这些人识别的BEA的cDNA 用于确定最适合作为靶点的表位结构的抗体 好的。 4.确定与细胞相关的BEA的最佳分子结构 以RIT为靶点，通过表位映射，构建嵌合重组 抗原和寡糖的分析，并通过体外和体内 (2)临床前评估。 某些抗乳房粘蛋白和46 kDa乳房抗原的单抗(BA46) 在RIT中有效。因为乳房的加工过程发生了变化 串联重复结构域中的一些表位优先 在乳腺癌中表达，因此某些识别这些的MoAbs 通常情况下，隐蔽表位在成像中比其他表位更有效 乳腺癌患者的转移。另外，对于BA46抗原，一些MoAbs 在临床前RIT研究中，抗它药比其他药物更有效。这 可能与该抗原可能的自分泌/旁分泌功能有关 参与细胞相互作用，以及不同的表位结构域 分子。项目2中制备的人MoAbs可识别表位 这后两个抗原，如果是这样的话，将确定准确的表位。 如果发现新的抗原，将对其进行分离和鉴定， 并对它们的cDNA进行克隆和测序。表位的分析将由 表位定位、寡糖分析和重组生产 抗原及其片段作为嵌合蛋白。分子 表位的特征将与治疗相关 体外和体内的有效性(项目2)临床前研究， 目的为放射免疫治疗选择最佳的靶位表位和单抗。

The overall goal of the two projects participating in this is to develop and test new monoclonal antibody based strategies for treatment of breast cancer. This project will explore the molecular and cell biology of breast epithelial antigens (BEAs) and their epitope structures that are the best targets for breast tumor localization for radioimmunotherapy (RlT). Project 2 aims to identify human B-lymphocytes recognized by the breast tumor in its human host, produce human monoclonal antibodies secreted by these cells by establishing stable cell lines producing them through genetic engineering, and to develop new strategies for radioimmunotherapy by humanization and fragmentation of existing MoAbs for translation into clinical trials. The SPECIFIC AIMS of this project are: 1. Isolate and characterize cell-associated forms of the breast mucin and BA46 and distinguish them from secreted forms using the collection of MoAbs that we have already developed. 2. Isolate and characterize the BEA identified by the human MoAbs developed in Project 2. Distinguish cell-associated and secreted forms. 3. Isolate and sequence the cDNAs for the BEA recognized by these human MoAbs for determining the epitope structures that are the best target for RIT. 4. Determine molecular structures of cell-associated BEA that are the best target for RIT, by epitope mapping, construction of chimeric recombinant antigens and oligosaccharide analysis, and by in vitro and in vivo (Project 2) preclinical evaluation. Certain MoAbs against the breast mucin and a 46 kDa breast antigen (BA46) are effective in RIT. Because of the altered processing of the breast mucin some epitopes In the tandem repeat domain are preferentially expressed in breast carcinomas, and thus certain MoAbs recognizing these normally cryptic epitopes are more effective than others in imaging metastases in breast cancer patients. Also, for BA46 antigen, some MoAbs against it are more effective than others in preclinical RIT studies. This may be related to this antigen's possible autocrine/paracrine function and involvement in cell interaction, and the different epitope domains of the molecule. The human MoAbs prepared in Project 2 may identify epitopes on these latter two antigens, If so, the precise epitopes will be determined. If new antigens are identified they will be isolated and characterized, and their cDNAs cloned and sequenced. The epitopes will be analyzed by epitope mapping, oligosaccharide analysis, and production of recombinant antigens and their fragments as chimeric proteins. Molecular characteristics of the epitopes will be correlated with therapeutic effectiveness in in vitro and in vivo (Project 2) preclinical studies, in order to select the best target epitope and MoAb for radioimmunotherapy.