Use of aAPC for melanoma adoptive immunotherapy

aAPC 用于黑色素瘤过继免疫治疗

• 批准号: 6810352
• 负责人: JONATHAN P SCHNECK
• 金额: $ 33.52万
• 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 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6810352
• 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; neoplasm /cancer therapy; 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）：虽然收养免疫疗法有望成为恶性黑色素瘤的一种治疗方法，但由于缺乏可再现且经济的方法来产生治疗数量的抗原特异性CTL，这阻碍了发展。我们的初步数据表明，通过将HLA-Ig和抗CD28耦合到珠子制造的人造抗原呈递细胞（AAPC）可以可靠地诱导和扩展从健康的供体到各种不同抗原的抗原特异性供体。这增加了HLA-Ig可用于替代基于标准的树突状细胞的基于抗原特异性CTL的非标准细胞的扩展。因此，存在使用基于HLA-Ig的AAPC作为临床级黑色素瘤特异性T细胞诱导，扩展和激活转移性黑色素瘤的可行方法。 在这项研究中，我们建议证明基于HLA-Ig的AAPC在诱导和扩展转移性黑色素瘤患者的抗肿瘤特异性CTL方面的功能疗效。具体而言，我们建议通过优化AAPC结构和刺激持续时间来优化AAPC刺激MART-1特异性CTL。 T细胞激活需要通过T细胞受体（信号1）和通过共刺激分子（信号2）进行信号的组合，例如B7 CD28的参与度。 AAPC的各种配方的功效将确定信号1与信号2以及不同类型的信号2的可变比率。将研究体外扩展的MART1特异性CTL，以在鼠模型中进行体内功能，包括人/SCID模型，以及使用采用转移的CTL的鼠A2-转基因小鼠。 CTL处理的功效可能会通过转移针对多个抗原表位的CTL种群来增强。因此，我们将分析诱导/扩展针对与黑色素瘤相关的A2限制亚辅助表位GPL00，NY-ESO-1和酪氨酸酶的CTL的能力。通过还具有黑色素瘤特异性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II类HLA的生物素化的AAPC，并生成基于II-LG类的AAPC，以刺激黑色素瘤特异性II类限制的CD4 T细胞，并分析在体内模型中具有黑色素瘤特异性CD8和CD4 T细胞的重要性。这些研究将有助于评估AAPC作为用于治疗转移性黑色素瘤的收养免疫治疗方法的潜在方法。