ANALYSIS OF E7 SPECIFIC T CELLS USING MHC-1GG MOLECULES

使用 MHC-1GG 分子分析 E7 特异性 T 细胞

• 批准号: 6511314
• 负责人: DREW M. PARDOLL
• 金额: $ 32.7万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-15 至 2003-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6511314
• 关键词: MHC class I antigen; MHC class II antigen; antineoplastics; biopsy; cervix neoplasms; chimeric proteins; clinical research; cytokine; cytotoxic T lymphocyte; dimer; drug screening /evaluation; female; flow cytometry; helper T lymphocyte; human papillomavirus; human subject; human therapy evaluation; immune tolerance /unresponsiveness; immunocytochemistry; immunoglobulin G; neoplasm /cancer immunotherapy; neoplasm /cancer vaccine; pharmacokinetics; tissue /cell culture; virus antigen

DESCRIPTION: (Applicant's Abstract) This application seeks to evaluate a novel vaccine approach involving targeting the HPV-16 E7 antigen to the MHC class II processing pathway. One of the most important endeavors in evaluating therapeutic vaccines is the measurement of induced in vivo immune responses relevant to anti-tumor efficacy. Recent studies call into question the sensitivity of standard bulk and limiting dilution CTL analyses in estimating the true frequency and functional status of antigen specific T cells. An exciting new approach to directly visualize antigen specific T cells has been the use of multimeric MHC+ peptide complexes that bind stably and specifically to antigen specific T cells. The applicant has developed a simple and versatile approach to MHC-peptide multimerization by linking MHC molecules genetically to a dimeric IgG scaffold. These peptide-MHC-IgG chimeras have been shown to bind stably and specifically to antigen-specific CD8+ and CD4+ T cells. He proposes to utilize these reagents to evaluate the in vivo dynamics and functional status of E7 specific T cells in patients receiving the LAMP targeted E7 vaccines. Specifically, he proposes to: 1) Develop stable CD8+ T cell lines and clones specific for immunodominant HLA-A2 restricted E7 peptides. 2) Develop and test E7-HLA-A2-IgG dimers for three identified immunodominant HLA-A2 restricted E7 peptides. 3) Evaluate the in vivo dynamics of E7+A2 specific CD8+ T cell responses in vaccinated HLA-A2+ patients utilizing the E7-HLA-A2-IgG chimeras developed in Specific Aim 2. 4) Develop analogous E7-MHC II-IgG chimeras capable of detecting CD4+ T cells specific for E7 peptides presented by the common HLA Class II alleles DR1 and DR4. Ultimately, these analyses will test the hypothesis that vaccination of patients expressing HPV 16 E7+ SIL or cervical cancer with LAMP-targeted E7 vaccines increases the numbers of activated E7 specific T cells in peripheral blood and at the site of disease.

描述：(申请者摘要)本申请旨在评估一部小说 将HPV-16 E7抗原靶向MHC II类的疫苗方法 加工途径。评估中最重要的努力之一 治疗性疫苗是对体内诱导的免疫反应的测量 与抗肿瘤疗效相关。最近的研究使人们对 标准体积和极限稀释度CTL分析在估算中的灵敏度 抗原特异性T细胞的真实频率和功能状态。一个 一种令人兴奋的直接显示抗原特异性T细胞的新方法已经出现 稳定和特异结合的多聚体MHC+肽复合体的应用 抗原特异性T细胞。申请人开发了一种简单而通用的 通过将MHC分子基因连接到MHC分子实现MHC多肽多聚化的研究 一种二聚体免疫球蛋白支架。这些多肽-MHC-免疫球蛋白嵌合体已被证明与 稳定和特异地结合抗原特异性的CD8+和CD4+T细胞。他提议 利用这些试剂评价体内动力学和功能 接受LAMP靶向E7的患者的E7特异性T细胞状态 疫苗。具体地说，他建议：1)建立稳定的CD8+T细胞系，并 针对免疫优势的人类白细胞抗原A2限制性E7多肽的克隆。2)发展 并检测E7-HLAA2-Ig G二聚体对三种已确定的免疫优势的HLAA2 限制性E7多肽。3)评价E7+A2特异性CD8+细胞的体内动力学 利用E7-HLAA2-Ig G免疫接种HLAA2+患者的T细胞反应 为特定目的而开发的嵌合体2)形成类似的E7-MHC II-Ig G 能检测E7多肽特异性CD4+T细胞的嵌合体 由常见的人类白细胞抗原II类等位基因DR1和DR4决定。最终，这些分析将 测试HPV16E7+sIL或表达HPV16E7+sIL的患者接种疫苗的假设 使用LAMP靶向E7疫苗的宫颈癌患者数量增加 激活外周血和发病部位的E7特异性T细胞。