IMMUNIZATION OF MELANOMA PATIENTS WITH GANGLIOSIDES

三种在黑色素瘤上表达的细胞表面抗原现在可在 纯化形式的神经节苷脂GD2、GD3和GM2。 三是作为 神经外胚层不同细胞亚群的分化抗原 脉 已知GD2和GM2在人体内具有潜在免疫原性 因为它们已经被各种人类血清所鉴定。 临床 GD3的相关性已经通过炎症反应和主要的 3例接受抗GD3单克隆抗体治疗的黑色素瘤患者的临床反应 抗体（R24）。 然而，我们一直无法持续发展 在人或小鼠中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，我们探讨了 纯化的GM2和GD2疫苗在小鼠中的作用，并已确定 持续诱导抗体反应的方法。 的成功 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个试验中，没有6个病人 用GM_2单独免疫的11例患者中有5例产生抗体， 用GM 2加BCG或GM 2加沙门氏菌突变体R595产生了 与GM2反应的抗体（中位滴度320）。 这些血清介导 补体依赖性对人黑色素瘤和星形细胞瘤细胞的细胞毒作用 人的补充。 ITLC证实，它们只与 GM 2。这里提出的方法是（并将继续）基于 这些正在进行的小鼠研究的结果。 在初步试验中， 将改变GM2加BCG的剂量、时间表和给药途径 或R595或两者以进一步增加抗GM 2应答率。 后续 试验将测试其他疫苗，如含有GM 2和GM 2的脂质体 共价连接至BSA或KLH。 II期黑色素瘤患者将 在淋巴结切除术后或前后接种疫苗， 他们的血清学反应和迟发型超敏反应， 相关神经节苷脂测试。 手术时切除的免疫淋巴结 将用于生产人类单克隆抗体。 方法挑衅 GD2和GD3将使用一致的血清学或DTH应答， 随后与三种神经节苷脂一起。 疫苗生产 II期患者的一致缓解将用于以下患者： 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.