IMMUNIZATION OF MELANOMA PATIENTS WITH GANGLIOSIDES

在黑色素瘤上表达的三种细胞表面抗原现在可以在 纯化后的神经节苷脂GD2、GD3和GM2。这三个都是 神经外胚层不同细胞亚群的分化抗原 血统。已知GD2和GM2在人类中都具有潜在的免疫原性 因为它们已经被各种人类血清所鉴定。临床部 GD3的相关性已被炎症反应和主要 抗GD3单抗治疗3例黑色素瘤患者的临床疗效 抗体(R24)。然而，我们一直无法持续发展 人或小鼠使用全细胞或细胞的免疫原性疫苗 表达这些神经节苷脂的裂解物。因此，我们探索了 纯化的GM2和GD2疫苗对小鼠的免疫效果及鉴定 持续诱导抗体反应的方法。的成功之处 这些小鼠试验鼓励我们继续进行类似的试验 II期黑色素瘤患者。在我们最初的3次试验中，6名患者 仅用GM2免疫产生抗体，但11例患者中有5例免疫 GM2+卡介苗或GM2+明尼苏达沙门氏菌。突变体R595已经产生了 抗体与GM2反应(中位滴度320)。这些血清可以调节 补体依赖性细胞毒作用对人黑色素瘤和星形细胞瘤细胞的影响 与人类相辅相成。ITLC证实，他们的反应仅限于 GM2。这里提出的方法是(并将继续)基于 在老鼠身上进行的这些研究的结果。在初审中，我们 将改变GM2加卡介苗的剂量、给药程序和给药途径 或R595或两者合用，以进一步提高抗GM2应答率。后续 试验将测试其他疫苗，如含有GM2和GM2的脂质体 与牛血清白蛋白或KLH共价连接。第二期黑色素瘤患者将 在淋巴清扫后或之前和之后接种疫苗 它们的血清学反应性和迟发性超敏反应 相关神经节苷脂检测。免疫淋巴结术中切除 将被用来生产人类的单抗。引发争议的方法 GD2和GD3将使用一致的血清学或DTH反应， 并随后与三种神经节苷脂一起。疫苗生产 II期患者的一致反应将用于以下患者 可测量的疾病，以衡量它们对免疫反应的影响 更严重的疾病。

Three cell surface antigens expressed on melanomas are now available in purified form, the gangliosides GD2, GD3 and GM2. All three serve as differentiation antigens for different subsets of cells of neuroectodermal lineage. Both GD2 and GM2 are known to be potentially immunogenic in man as they have been identified by a variety of human sera. The clinical relevance of GD3 has been demonstrated by inflammatory reactions and major clinical responses in 3 melanoma patients treated with anti-GD3 monoclonal antibody (R24). We have however, been unable to develop consistently immunogenic vaccines in man or in the mouse using whole cells or cell lysates expressing these gangliosides. Consequently, we have explored the effect of purified GM2 and GD2 vaccines in the mouse and have identified approaches that consistently induce an antibody response. The success of these murine trials has encouraged us to proceed with similar trials in Stage II melanoma patients. In our initial 3 trials, none of 6 patients immunized with GM2 alone produced antibody but 5 of 11 patients immunized with GM2 plus BCG or GM2 plus Salmonella minn. mutant R595 have produced antibody reactive with GM2 (median titer 320). These sera mediate complement dependent cytotoxicity on human melanoma and astrocytoma cells with human complement. ITLC confirmed that they react exclusively with GM2. The approaches proposed here are (and will continue to be) based on the results of these ongoing studies in the mouse. In intitial trials we will change the dose, schedule and route of administration of GM2 plus BCG or R595 or both to further increase the anti GM2 response rate. Subsequent trials will test other vaccines such as liposomes containing GM2 and GM2 covalently attached to BSA or KLH. Stage II melanoma patients will be vaccinated after, or before and after, lymphadenectomy in small groups and their serological reactivity and delayed hypersensitivity reactions to the relevant gangliosides tested. The immunized lymph nodes removed at surgery will be used to produce human monoclonal antibodies. Approaches provoking a consistent serological or DTH responses will be used with GD2 and GD3, and subsequently with the three gangliosides together. Vaccines producing a consistent response in Stage II patients will be used in patients with measurable disease to gauge their effect on immune responses in the face of more advanced disease.