Identification of Immune Selected Breast Cancer Antigens

免疫选择乳腺癌抗原的鉴定

• 批准号: 7119620
• 负责人: Kevin P. Claffey
• 金额: $ 14.16万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-06 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7119620
• 关键词: B lymphocyte; antigen antibody reaction; antitumor antibody; binding proteins; biomarker; breast neoplasms; female; gene mutation; genetic library; high throughput technology; human tissue; humoral immunity; immune response; leukocyte activation /transformation; liquid chromatography mass spectrometry; lymph nodes; lymphocyte proliferation; method development; neoplasm /cancer immunology; recombinant proteins; tumor antigens

DESCRIPTION (provided by applicant): Immune-dependent responses are selective in defining non-self or aberrant antigen presentation. Unfortunately, long-term antibody production to human cancer antigens is limited. In an innovative and novel approach, we have developed a method to utilize primary immune reactions in tumor draining lymph nodes to provide the means to define biologically active tumor antigens originating from breast cancers. This novel approach combines a series of steps to coordinate the construction of low complexity antibody cDNA libraries and protein production that are used to identify tumor antigens using sensitive antibody microscale "antigen-trap" assays followed by LC-MS/MS antigen identification. Tumor antigens identified can then be verified as potential tumor antigens using biochemical, immunological and molecular methodologies. The methodologies applied are medium throughput platform based designed to rapidly evaluate matched lymph node and tumor samples from the same patient. This project applies innovative technologies that demonstrate that: a) tumor draining lymph nodes are immuno-reactive to aberrant breast cancer antigens and produce antigen-dependent somatic hypermutation in proliferative B-cell germinal centers, b) antigen binding domains of somatic hypermutated antibodies synthesized as recombinant VH and/or VHVl/Vk ScFv proteins can specifically recognize and identify breast cancer antigens, and c) antigens identified can be verified as diagnostic for breast cancer sub-phenotypes. This R21 application proposes to expand and refine our methodologies to: 1) determine the diversity and effectiveness of immune-selection of tumor antigens in a larger patient population, 2) expand our ability to produce antibody molecules with appropriate structure and antigen binding, and 3) develop methodologies to incorporate antibody proteins synthesized into highly sensitive assays that can screen primary cancer, histological material and/or biological fluids necessary to evaluate the potential of antigens as diagnostic biomarkers.

描述（由申请人提供）： 免疫依赖性反应在定义非自身或异常抗原呈递方面具有选择性。不幸的是，针对人类癌症抗原的长期抗体生产是有限的。通过一种创新和新颖的方法，我们开发了一种利用肿瘤引流淋巴结中的初级免疫反应的方法，以提供定义源自乳腺癌的生物活性肿瘤抗原的方法。这种新颖的方法结合了一系列步骤来协调低复杂性抗体 cDNA 文库的构建和蛋白质生产，这些步骤用于使用敏感的抗体微量“抗原捕获”测定来鉴定肿瘤抗原，然后进行 LC-MS/MS 抗原鉴定。然后可以使用生化、免疫学和分子方法将鉴定出的肿瘤抗原验证为潜在的肿瘤抗原。所采用的方法是基于中等通量平台的，旨在快速评估来自同一患者的匹配淋巴结和肿瘤样本。该项目应用创新技术证明：a) 肿瘤引流淋巴结对异常乳腺癌抗原具有免疫反应性，并在增殖性 B 细胞生发中心产生抗原依赖性体细胞超突变，b) 合成的体细胞超突变抗体的抗原结合域重组VH和/或VHV1/Vk ScFv蛋白可以特异性识别和鉴定乳腺癌抗原，并且c)所鉴定的抗原可以被验证为乳腺癌亚表型的诊断。该 R21 申请建议扩展和完善我们的方法，以：1) 确定更大患者群体中肿瘤抗原免疫选择的多样性和有效性，2) 扩大我们生产具有适当结构和抗原结合的抗体分子的能力，以及 3 ）开发方法，将合成的抗体蛋白纳入高度灵敏的检测中，以筛选原发性癌症、组织学材料和/或评估抗原作为诊断生物标志物潜力所需的生物液体。

项目成果

Identification of novel tumor antigens with patient-derived immune-selected antibodies.

使用患者来源的免疫选择抗体鉴定新型肿瘤抗原。

• 发表时间: 2009-02
• 作者: Rodriguez;Sparkowski, Jason;Keough, Martin P;Phoenix, Kathryn N;Vumbaca, Frank;Han, David K;Gundelfinger, Eckart D;Beesley, Philip;Claffey, Kevin P
• 通讯作者: Claffey, Kevin P