IMMUNIZATION OF MELANOMA PATIENTS WITH GANGLIOSIDES

四种神经节苷脂GM2、GD2、GD3和9-0-乙酰GD3，以及高 分子量黑色素瘤硫酸软骨素蛋白多糖表位 (MEL-CSPG)是黑色素瘤的主要细胞膜成分，我们 已被选定为免疫控制的潜在目标 免疫接种。其中，GM2的免疫原性最强，但与GM2的反应性最强 在很大程度上被限制在IgM抗体的模式中 与GM2作为T细胞非依赖性抗原的作用一致。 因此，我们在鼠标中筛选了大量较新的 增强或替换T细胞帮助的方法。最多的三个 有希望的方法已被选为辅助临床试验#年 AJCC III期黑色素瘤患者：一种有效的新巨噬细胞激活剂和B 细胞有丝分裂原，称为蛋白质小体，作为佐剂/载体单独使用或 与其他佐剂或有丝分裂原混合；高共价连接 神经节苷脂对KLH的表位密度用于诱导T细胞辅助；以及 抗独特型抗体的结合部位是 GD3和MEL-CSPG免疫原性表位的内部图像。 在免疫后，抗体反应将被监测 培养黑色素瘤细胞的ELISA、免疫染色和检测，以及迟发型变态反应 针对免疫原和相关表位的皮肤测试反应将是 量过了。我们的具体目标是： 1.确定和完善最有效的诱导方法 针对这5种抗原中至少3种的抗体和/或迟发型变态反应。 2.构建多价疫苗并检测其免疫原性 关于目标1中的发现，并根据剂量、路线对其进行优化 和日程安排。 3.在更大范围内测试这种明显具有免疫原性的多价疫苗 对患有可测量疾病的患者进行治疗试验。

Four gangliosides, GM2, GD2, GD3 and 9-0-acetyl GD3, and the high molecular weight melanoma chondroitin sulfate proteoglycan epitope (mel-CSPG) are prominent cell membrane components of melanoma that we have selected as potential targets for immunological control after active immunization. Of these, GM2 is most immunogenic, but reactivity with GM2 has been largely restricted to IgM antibodies in a pattern most consistent with GM2 acting as a T cell independent antigen. Consequently, we have screened in the mouse, a large number of newer methods for augmenting or replacing T cell help. The three most promising approaches have been selected for adjuvant clinical trials in AJCC Stage III melanoma patients: a potent new macrophage activator and B cell mitogen termed proteosomes, as adjuvant/vehicle used either alone or mixed with other adjuvants or mitogens; covalent attachment at high epitope density of ganglioside to KLH for induction of T cell help; and the use of anti-idiotypic antibodies with binding sites which are internal images of the immunogenic epitopes of GD3 and mel-CSPG. following immunization, the antibody response will be monitored with ELISAs, immune stains and assays on cultured melanoma cells, and the DTH skin test response against the immunogen and related epitopes will be measured. Our specific aims are: 1. Identify and perfect the approaches which most effectively induce antibodies and/or DTH against at least 3 of these 5 antigens. 2. Construct and test the immunogenicity of a polyvalent vaccine based on the findings in Aim 1 and optimize it with regard to dose, route and schedule. 3. Test this demonstrably immunogenic polyvalent vaccine in a larger therapy trial in patients with measurable disease.