RNA pulsed Dendritic Cells as Immunotherapy for Melanoma

RNA脉冲树突状细胞作为黑色素瘤的免疫疗法

• 批准号: 6514769
• 负责人: Matthew Frank Kalady
• 金额: $ 0.84万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6514769
• 关键词: RNA; cytotoxic T lymphocyte; dendritic cells; human subject; human therapy evaluation; melanoma; metastasis; neoplasm /cancer immunology; neoplasm /cancer immunotherapy; neoplasm /cancer vaccine; patient oriented research; transfection; tumor antigens; vaccine development

Malignant melanoma remains a formidable challenge to patients, physicians, and scientists. Although surgical resection serves as the primary therapy and is often curative, melanoma recurs both locally and at distant sites. Unfortunately, current adjuvant therapies have not improved the dismal prognosis associated with regional and metastatic disease. This research focuses on the use of immunotherapy to treat micrometastatic and recurrent melanoma. Information gained through this preclinical research will be applied to developing an effective clinical melanoma vaccine. Immunologic rejection of tumor cells is mediated by cytotoxic T-lymphocytes (CTL) which recognize specific tumor- associated antigens (TAA). Antigen presenting cells, such as dendritic cells (DC), have been shown to generate an antigen-immunologic response both in vitro and in vivo after priming with TAA. Current research and clinical trials are underway at this institution loading of DC with tumor lysates and TAA peptides. This research project will attempt to optimize the CTL response against melanoma by methodologically studying the methods used to prime DC with tumor RNA induce unexpectedly potent tumor-specific CTL responses. Specifically, DC will be isolated through leukopheresis from patients with melanoma in whom tumor cell lines can be established. The DC from these patients will be primed with total tumor RNA from the patients' own tumor, or primed with combinations of previously isolated RNA for melanoma-specific TAA such as MART-1, MAGE-3, tumor, or primed with combinations of previously isolated RNA for melanoma-specific TAA such as MART-1, MAGE-3, tumor, or primed with combinations of previously isolated RNA for melanoma-specific TAA such as MART-1, MAGE-3, tyrosinase, and gp100. The primed dendritic cells will be incubated with CTL. These CTL will subsequently be mixed with melanoma cell lines and the percentage of lysed cells will be quantified using a chromium release assay. The antigen that yields the greatest cytoxicity will then by directly tested against the current method of pulsing dendritic cells with protein yields the greater cytotoxicity will then be directly tested against the current method of pulsing dendritic cells with protein lysates and peptides. Data gained from this preclinical work will guide future uses of vaccine development in the clinic.

恶性黑色素瘤对患者、医生和科学家来说仍然是一个巨大的挑战。尽管手术切除是主要治疗方法并且通常可以治愈，但黑色素瘤会在局部和远处复发。不幸的是，目前的辅助治疗并没有改善与局部和转移性疾病相关的不良预后。这项研究的重点是使用免疫疗法来治疗微转移和复发性黑色素瘤。通过这项临床前研究获得的信息将用于开发有效的临床黑色素瘤疫苗。肿瘤细胞的免疫排斥是由识别特定肿瘤相关抗原（TAA）的细胞毒性T淋巴细胞（CTL）介导的。抗原呈递细胞，如树突状细胞 (DC)，已被证明在用 TAA 引发后在体外和体内产生抗原免疫反应。该机构目前正在进行用肿瘤裂解物和 TAA 肽加载 DC 的研究和临床试验。该研究项目将尝试通过方法学研究用肿瘤 RNA 启动 DC 的方法来优化针对黑色素瘤的 CTL 反应，从而诱导出乎意料的有效的肿瘤特异性 CTL 反应。具体来说，DC将通过白细胞去除术从可以建立肿瘤细胞系的黑色素瘤患者中分离出来。来自这些患者的DC将用来自患者自身肿瘤的总肿瘤RNA引发，或用先前分离的用于黑色素瘤特异性TAA（例如MART-1、MAGE-3、肿瘤）的RNA组合引发，或用先前分离的用于黑色素瘤特异性TAA（例如MART-1、MAGE-3、肿瘤）的RNA组合引发，或用先前分离的用于黑色素瘤特异性TAA（例如）的RNA组合引发。 MART-1、MAGE-3、酪氨酸酶和 gp100。引发的树突状细胞将与 CTL 一起孵育。这些 CTL 随后将与黑色素瘤细胞系混合，并使用铬释放测定对裂解细胞的百分比进行定量。然后将产生最大细胞毒性的抗原直接针对当前用蛋白质脉冲树突细胞的方法进行测试，产生更大的细胞毒性然后将直接针对当前用蛋白质裂解物和肽脉冲树突细胞的方法进行测试。从这项临床前工作中获得的数据将指导未来疫苗开发在临床中的使用。