GANGLIOSIDE GD2 AS TARGET FOR IMMUNOTHERAPY IN MELANOMA

这项建议的总体目标是应用抗独特型（ld） 基于疫苗的方法治疗人类黑色素瘤。 二唾液酸神经节苷脂GD 2在黑素瘤细胞上以高密度表达， 将被用作免疫治疗的靶点 我们将启动第1b阶段 使用命名为1A 7（IgG 1-k）的抗id抗体的临床试验， 在功能上模拟GD 2。 小鼠单克隆抗GD 2抗体14 G2 a被免疫组化。 用作免疫抗体或Ab 1，抗Id 1A 7（或Ab 2） 产生了。 1A 7用于诱导抗GD 2特异性抗体（Ab 1 '） 在老鼠兔子和猴子身上 在这项临床研究中，我们将使用抗ld 1A 7与QS-21佐剂混合以主动免疫GD 2阳性 黑色素瘤患者。 本试验的目的是1）确定 1A 7对患者产生Ab 3（Ab 1 ′）应答的能力的影响 和对自体和/或同种异体特异性的细胞溶解性T淋巴细胞（CTL）， 肿瘤细胞; 2）评估1A 7的毒性; 3）确定1A 7的毒性。 抗ld 1A 7的最佳免疫调节剂量;和4）监测 临床反应。 在Ib期试验完成时，我们将开始 使用最佳免疫调节剂量的II期试验。 我们的目标将 还可以通过使用抗GD 2抗体增强动物模型中的抗GD 2免疫应答， 利用重组DNA技术构建基于1A 7结构的DNA疫苗。 这些DNA疫苗将是质粒和重组牛痘病毒， 1A 7片段在哺乳动物细胞中的表达。 1A 7片段将 由重链可变结构域组成的单链多肽， 和由15个氨基酸的接头连接的轻链。 治疗潜力 这些疫苗将通过测定体液和细胞 小鼠免疫应答并与标准1A 7-QS-21疫苗进行比较 以及抗原疫苗GD 2-KLH加QS-21。 所有这些疫苗 将测试肿瘤保护和已建立肿瘤的治疗， 免疫活性鼠淋巴瘤模型，由表达 GD 2高密度。 这项研究将是一项临床试验的前奏， 黑色素瘤患者使用第二代基于抗Id的DNA疫苗。

The overall objective of this proposal is to apply an anti-idiotype(ld) based vaccine approach for the treatment of human melanoma. Disialoganglioside GD2 is expressed at high density on melanoma cells and will be used as a target for immunotherapy. We will initiate a Phase 1b clinical trial with an anti-id antibody, designated 1A7 (lgG1-k), which functionally mimics GD2. Murine monoclonal anti-GD2 antibody 14G2a was used as the immunizing antibody or Ab1, against which anti-ld 1A7 (or Ab2) was generated. 1A7 was used to induce anti-GD2 specific antibodies (Ab1') in mice, rabbits, and monkeys. In this clinical study, we will use anti-ld 1A7 mixed with the QS-21 adjuvant to actively immunize GD2 positive melanoma patients. The objectives of this trial are 1) to determine the effects of 1A7 on the ability of patients to generate Ab3 (Ab1') responses and cytolytic T lymphocytes (CTL) specific for autologous and/or allogeneic tumor cells; 2) to evaluate the toxicity of 1A7; 3) to determine the optimal immunomodulatory dose of the anti-ld 1A7; and 4) to monitor for clinical responses. At the completion of the Phase lb trial, we will begin a Phase II trial using the optimum immunomodulatory dose. Our goal will also be to enhance in animal models the anti-GD2 immune responses by using DNA vaccines based on the structure of 1A7 by recombinant DNA technology. These DNA vaccines will be plasmids and recombinant vaccinia virus capable of expression of 1A7 fragments in mammalian cells. 1A7 fragments will be a single chain polypeptide consisting of the variable domains of the heavy and light chains linked by a 15 amino acid linker. Therapeutic potential of these vaccines will be evaluated by determining the humoral and cellular immune responses in mice and compared with the standard 1A7-QS-21 vaccine as well s the antigen vaccine GD2-KLH plus QS-21. All of these vaccines will be tested for tumor protection and therapy of established tumors in an immunocompetent murine lymphoma model consisting of EL4 cells which express GD2 at high density. This study will be prelude to a clinical trial for melanoma patients with second generation anti-ld based DNA vaccines.