Use of aAPC for melanoma adoptive immunotherapy

aAPC 用于黑色素瘤过继免疫治疗

• 批准号: 7099559
• 负责人: JONATHAN P SCHNECK
• 金额: $ 32.73万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7099559
• 关键词: CD28 molecule; CD4 molecule; MHC class II antigen; SCID mouse; antibody; biological signal transduction; clinical research; cytotoxic T lymphocyte; genetically modified animals; helper T lymphocyte; human subject; immunofluorescence technique; immunologic substance development /preparation; laboratory mouse; leukocyte activation /transformation; melanocyte; melanoma; neoplasm /cancer immunotherapy; passive immunization; therapy design /development; tumor antigens

DESCRIPTION (provided by applicant): While adoptive immunotherapy holds promise as a treatment for malignant melanoma, development has been impeded by the lack of a reproducible and economical method for generating therapeutic numbers of antigen-specific CTL. Our preliminary data demonstrate that an artificial Antigen-Presenting Cell (aAPC), made by coupling HLA-Ig and anti-CD28 to beads, can reliably induce and expand antigen-specific CTL from healthy donors to a variety of different antigens. This raises the possibility that HLA-Ig can be used to replace standard dendritic cell based ex vivo expansion of antigen-specific CTL. Therefore the potential exists to use HLA-Ig-based aAPC as a viable method for induction, expansion and activation of clinical grade melanoma-specific T cells in the treatment of metastatic melanoma. In this study we propose to demonstrate functional efficacy of HLA-Ig based aAPC for inducing and expanding anti-tumor specific CTL from patients with metastatic melanoma. Specifically we propose to optimize aAPC stimulation of Mart-1 specific CTL by optimizing aAPC structure and the duration of stimulation. T cell activation requires delivery of a combination of signals through the T cell receptor (Signal 1) and through co-stimulatory molecules (Signal 2) such as engagement of CD28 by B7. The efficacy of the various formulations of aAPC, with variable ratios of signal 1 to signal 2 as well as different type of signal 2, will be determined. In vitro expanded Mart1-specific CTL will be studied for in vivo function in murine models, including a human/SCID model and possibly the murine A2-transgenic mice using adoptively transferred CTL. Efficacy of CTL treatments is likely to be augmented by transferring CTL populations directed at multiple antigenic epitopes. Therefore we will analyze the ability to induce/expand CTL specific for the melanoma associated, A2-restricted subdominant epitopes gpl00, NY-ESO-1 and tyrosinase. Effective immunotherapy will be augmented by also having melanoma-specific CD4 T cells. Our preliminary data indicates that using biotinylated class II-based aAPC or autologous DC, we can generate gpl00 melanoma specific CD4 cells. We propose to further develop the biotinylated class II HLA-based aAPC and to generate class II-lg-based aAPC for stimulation of melanoma specific class II-restricted CD4 T cells and analyze the importance of having both melanoma specific CD8 and CD4 T cells in the in vivo models. These studies will help evaluate the role of aAPC as a potential approach to adoptive immunotherapeutic for the treatment of metastatic melanoma.

描述（由申请人提供）：虽然过继免疫疗法有望成为恶性黑色素瘤的治疗方法，但由于缺乏可重复且经济的方法来产生治疗数量的抗原特异性 CTL，其发展受到阻碍。我们的初步数据表明，通过将 HLA-Ig 和抗 CD28 与珠子偶联而制成的人工抗原呈递细胞 (aAPC) 可以可靠地诱导和扩展来自健康供体的抗原特异性 CTL 到各种不同的抗原。这提出了 HLA-Ig 可用于替代基于标准树突状细胞的抗原特异性 CTL 离体扩增的可能性。因此，有可能使用基于 HLA-Ig 的 aAPC 作为诱导、扩增和激活临床级黑色素瘤特异性 T 细胞来治疗转移性黑色素瘤的可行方法。 在这项研究中，我们打算证明基于 HLA-Ig 的 aAPC 在诱导和扩大转移性黑色素瘤患者的抗肿瘤特异性 CTL 方面的功能功效。具体来说，我们建议通过优化 aAPC 结构和刺激持续时间来优化 Mart-1 特异性 CTL 的 aAPC 刺激。 T 细胞激活需要通过 T 细胞受体（信号 1）和共刺激分子（信号 2）传递信号组合，例如 B7 与 CD28 的结合。将确定aAPC各种制剂的功效，其中信号1与信号2的比率可变以及信号2的类型不同。将在小鼠模型中研究体外扩增的 Mart1 特异性 CTL 的体内功能，包括人类/SCID 模型以及可能使用过继转移 CTL 的小鼠 A2 转基因小鼠。通过转移针对多个抗原表位的 CTL 群体，CTL 治疗的功效可能会得到增强。因此，我们将分析诱导/扩展对黑色素瘤相关的A2限制性亚优势表位gp100、NY-ESO-1和酪氨酸酶特异的CTL的能力。黑色素瘤特异性 CD4 T 细胞将增强有效的免疫疗法。我们的初步数据表明，使用生物素化的基于 II 类的 aAPC 或自体 DC，我们可以生成 gpl00 黑色素瘤特异性 CD4 细胞。我们建议进一步开发基于生物素化的 II 类 HLA 的 aAPC，并生成基于 II 类-lg 的 aAPC，用于刺激黑色素瘤特异性 II 类限制性 CD4 T 细胞，并分析黑色素瘤特异性 CD8 和 CD4 T 细胞在体内模型。这些研究将有助于评估 aAPC 作为治疗转移性黑色素瘤的过继免疫疗法的潜在方法的作用。