SPECIFIC ACTIVE IMMUMOTHERAPY OF HUMAN MELANOMA

我们将制备新鲜人黑色素瘤细胞的正丁醇提取物， 用于主动特异性免疫治疗的同种异体“疫苗”。 的 每种提取物中的黑色素瘤相关抗原将通过一组 小鼠和人单克隆抗体，并通过结合定量 我们开发的抑制程序。 人单克隆抗体将 从人-小鼠和人-人融合中不断发展， 黑色素瘤患者的局部淋巴结细胞作为一个伴侣， 非分泌型小鼠（M5）骨髓瘤或人B或骨髓瘤细胞系作为免疫抑制剂。 另一个合伙人。 这些人类单克隆抗体将使我们能够 确定被患者识别为“外来”的表位的存在， 丁醇提取物用作免疫原。 毒性和免疫 3种抗原提取物的标准池的效果将在 在转移性黑色素瘤患者中进行的正式I期试验。 两倍 将在每组中进行升级，共3个水平， 剂量，有和没有低剂量环磷酰胺作为增效剂， 免疫力 除了注意标准毒性，我们还将测量 抑制性T细胞通过各种互补方法，以确定 这种潜在有害后果的可能发生， 次免疫 几种测试抗体黑色素瘤，包括免疫 粘附、蛋白A混合红细胞吸附和两项嗜细胞性试验 抗体（LAI和ADCC）。 我们还开发了一种新的 基于单克隆抗体竞争性抑制的滴定方法 结合，灵敏度为1-2 ng抗体，以测量大多数 敏感地检测患者的血清抗体反应。 延迟 超敏反应将通过皮肤试验与黑色素瘤池进行研究 萃取物 我们还将探讨有限稀释分析培养T 细胞作为检测细胞溶解性抗黑色素瘤T细胞的新手段， 细胞介导的免疫的其它测试不一致地可靠。 该提案的发展方面，提高竞争力， 结合，有限稀释分析和抑制细胞方法，事实上， 这是整个研究的重要基础。 这么近 监测的临床试验可能有助于确定最重要的抗原， 包括在“疫苗”中，导致它们在未来研究中有选择地使用， 特异性主动免疫治疗黑色素瘤。

We will prepare 1-butanol extracts of fresh human melanoma cells to use as an allogeneic "vaccine" for active specific immunotherapy. The melanoma-associated antigens in each extract will be detected by a panel of mouse and human monoclonal antibodies and quantitated by a binding inhibition procedure we have developed. Human monoclonal antibodies will be continually developed from human-mouse and human-human fusions, with regional lymph node cells from melanoma patients as one partner and a nonsecretory mouse (M5) myeloma or a human B or myeloma cell line as the other partner. These human monoclonal antibodies will enable us to determine the presence of epitopes recognized by patients as "foreign", in the butanol extracts used as immunogens. The toxicity and immunological effects of a standard pool of 3 antigenic extracts will be tested in a formal Phase I trial, with metastatic melanoma patients. Two-fold escalations will be performed in each group for a total of 3 levels of dosage, with and without low-dose cyclophosphamide as a potentiator of immunity. Besides noting standard toxicity, we will also measure suppressor T cells by various complementary methods, to determine the possible occurence of this potentially harmful consequence of immunization. Several tests for antibodies to melanoma including immune adherence, protein A mixed hemadsorption and two tests for cytophilic antibodies (LAI and ADCC), will be performed. We have also developed a new titration method based on competitive inhibition of monoclonal antibody binding, with a sensitivity of 1-2 ng of antibody, to measure most sensitively the serum antibody response in the patients. Delayed hypersensitivity will be studied by skin testing with the pool of melanoma extracts. We will also explore limiting dilution analysis of cultivated T cells as a new means of detecting cytolytic antimelanoma T cells, since other tests of cell-mediated immunity have been inconsistently reliable. The developmental aspects of this proposal, with improvement of competitive binding, limiting dilution analysis and suppressor cell methods, in fact constitute an important rationale for the entire study. This closely monitored clinical trial may help to define the most important antigens to include in a "vaccine", leading to their selective use in future studies of specific active immunotherapy for melanoma.