IMMUNIZATION OF MELANOMA PATIENTS WITH GANGLIOSIDES

四种神经节苷脂，GM 2，GD 2，GD 3和9-0-乙酰基GD 3，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， 选择作为免疫控制的潜在目标后， 次免疫 其中，GM 2是最具免疫原性的，但与GM 2的反应性 在很大程度上仅限于IgM抗体， 与GM 2作为T细胞非依赖性抗原的作用一致。 因此，我们在小鼠中筛选了大量更新的 增加或替代T细胞辅助的方法。 三个最 有前途的方法已被选为辅助临床试验， AJCC III期黑色素瘤患者：一种有效的新型巨噬细胞激活剂和B 称为蛋白体的细胞有丝分裂原，作为单独或 与其他佐剂或有丝分裂原混合;高浓度共价连接 神经节苷脂对KLH的表位密度，用于诱导T细胞辅助;和 使用具有结合位点的抗独特型抗体， GD 3和mel-CSPG的免疫原性表位的内部图像。 免疫后，将用以下方法监测抗体应答： 培养的黑色素瘤细胞的ELISA、免疫染色和测定，以及DTH 针对免疫原和相关表位的皮肤试验应答将 测定了 我们的具体目标是： 1. 确定并完善最有效地诱导 针对这5种抗原中至少3种的抗体和/或DTH。 2. 构建并测试基于多价疫苗的免疫原性 根据目标1中的发现，并在剂量、途径 和时间表。 3. 在更大的实验室中测试这种明显具有免疫原性的多价疫苗。 在患有可测量疾病的患者中进行治疗试验。

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.