MELANOMA ANTIGENS INDENTIFIED FROM GMCSF VACCINATION

从 GMCSF 疫苗接种中鉴定出黑色素瘤抗原

• 批准号: 6522454
• 负责人: FRANK S HODI
• 金额: $ 8.74万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2003-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6522454
• 关键词: T lymphocyte; autologous transplantation; clinical research; colony stimulating factor; human subject; melanoma; neoplasm /cancer transplantation; neoplasm /cancer vaccine; neoplastic cell; tumor antigens; vaccine development

The ability to manipulate genes encoding potential immunomodulators such as cytokines and tumor antigens has opened a new era of research attempts. Poorly immunogenic murine tumor model systems demonstrate that vaccination with irradiated tumor cells engineered to secrete granulocyte-macrophage colony stimulating (GM-CSF) generates potent, specific, and long-lasting immunity. To test this strategy in patients with advanced melanoma, the laboratory of the applicants performed a clinical trial of vaccination with lethally irradiated, autologous melanoma cells expressing human GM-CSF. Vaccination elicits a striking infiltrate of pre-existing metastatic lesions with large numbers of lymphocytes, plasma cells, macrophages, and eosinophils, resulting in tumor destruction, fibrosis, and edema. Consistent production of humoral and cellular immune responses is exemplified by tumor specific cytotoxicity and cytokine production characterizing tumor-infiltrating lymphocytes, and post-vaccination sera containing IgG antibodies that recognize intracellular and cell surface melanoma determinants. These results provide a solid foundation for undertaking a molecular analysis of the melanoma antigens stimulating specific T and B cell responses in vaccinated patients. A number of candidate antigens have been identified from initial screening with autologous sera of a cDNA expression library constructed from a patient's tumor cell line. The first revealed a novel product with structural similarities to MUC-1, a member of a family of immunogenic cell surface molecules expressed in a variety of cancers. This antibody-based strategy will likely be productive for identifying a number of melanoma antigens. The applicants have designed several approaches to characterize immune responses against each candidate antigen. Humoral responses will be evaluated by immunoblotting lysates of CRIP packaging cell lines transfected with MFG based retroviral vectors expressing each candidate. Autologous EBV transformed lymphoblasts transfected with MFG retroviruses will be used to determine T cell cytotoxicity, proliferation, and cytokine production in response to each candidate antigen. Expression patterns will be determined by Northern analyses and RT-PCR from a variety of tumors and their normal tissue counterparts. Attempts by numerous investigators to utilize previously identified antigens in vaccine strategies demonstrate an ability to develop significant immune responses against these antigens. The analysis of the applicants should contribute to the understanding of the relative importance of common versus unique targets in anti-tumor immunity. Further identification of immunogenic antigens in melanoma offers promise for future therapies.

操纵编码潜在免疫调节剂基因的能力 随着细胞因子和肿瘤抗原的发展，新的研究时代 尝试。 免疫原性鼠肿瘤模型系统证明了 用辐照的肿瘤细胞进行疫苗接种，以分泌为分泌 粒细胞巨噬细胞刺激（GM-CSF）产生有效的 特异性和持久的免疫力。 在患者中测试这种策略 与晚期黑色素瘤一起，申请人的实验室进行了 疫苗接种的临床试验，用致命的自体辐照 表达人GM-CSF的黑色素瘤细胞。 疫苗接种引起了惊人的 浸润已有大量转移性病变 淋巴细胞，浆细胞，巨噬细胞和嗜酸性粒细胞，导致 肿瘤破坏，纤维化和水肿。稳定的体液产生 细胞免疫反应以肿瘤特异性为例 细胞毒性和细胞因子的产生，表征肿瘤浸润 淋巴细胞和疫苗后血清中含有IgG抗体 识别细胞内和细胞表面黑色素瘤决定因素。 这些 结果为进行分子分析提供了坚实的基础 黑色素瘤抗原刺激特定T和B细胞反应的抗原 接种疫苗的患者。 许多候选抗原已经 从cDNA的自体血清初始筛选中鉴定 表达式文库由患者的肿瘤细胞系构建。 这 首先揭示了一种与MUC-1结构相似的新型产品， 在以表达的免疫细胞表面分子家族的成员 各种癌症。 这种基于抗体的策略可能是 鉴定许多黑色素瘤抗原的生产力。 这 申请人设计了多种特征免疫的方法 针对每个候选抗原的反应。 体液反应将是 通过CRIP包装细胞系的免疫印迹裂解物评估 用表达每个候选者的基于MFG的逆转录病毒载体转染。 自体EBV转化了用MFG转染的淋巴细胞 逆转录病毒将用于确定T细胞细胞毒性， 响应每个候选者的增殖和细胞因子产生 抗原。 表达模式将通过北方分析确定 来自多种肿瘤及其正常组织的RT-PCR 同行。 众多调查人员以前使用的尝试 疫苗策略中确定的抗原证明了一种能力 针对这些抗原产生明显的免疫反应。 这 对申请人的分析应有助于理解 抗肿瘤中共同目标与独特目标的相对重要性 免疫。 黑色素瘤中免疫原性抗原的进一步鉴定 为将来的疗法提供希望。