Active Immune Therapy ot Leukemia Associated Antigens and Gene Therapy

用用编码鼠CD 154的腺病毒（Ad-154）转导的自体CLL细胞治疗的患者， CD 154）经历了白血病细胞计数和淋巴结大小的急性下降， 影响，包括白血病反应性T细胞数量增加，抗CLL抗体的产生， 疾病进展明显减缓或停止。接受治疗的患者中，旁观者未转染的 CLL细胞，反映了体内CD 154的活化。我们发现，CD 154激活诱导了一种新的免疫应答。 CLL细胞中的程序化系列事件，其最初被瞬时保护，然后被致敏。 通过连接诱导的外源性死亡受体（例如CD 95和DR 5）、细胞毒性药物或 抑制细胞凋亡抑制剂（IAP）蛋白的新药物（见项目2）。对这些药物敏感 似乎与CD 154激活诱导促凋亡分子表达的能力有关 投标通过p53独立，c-Abl依赖的机制，这显然涉及α亚型的 第73页。这为Ad-CD 154基因治疗可能避免对p53的依赖性提出了令人兴奋的前景 同时诱导抗白血病细胞和抗体免疫反应。后者 已经使我们能够鉴定一种新的CLL相关抗原Ror 1，它可能用于治疗 检测Ad-CD 154基因治疗的活性或用于该疾病的免疫治疗的疫苗。 对于未来的临床研究，我们开发了一种“人源化”CD 154（命名为ISF 35），其可以稳定地 在CLL质膜上高水平表达。本项目的具体目标如下：（1） 继续分析白血病细胞计数急性下降的机制 在用自体Ad-CD 154转导的或ISF 35转导的CLL细胞治疗的患者中观察到;（2） 检查针对新鉴定的CLL相关抗原Ror 1的细胞免疫应答;（3）评估 Ad-CD 154基因治疗诱导的抗白血病抗体的特异性和生物学活性， 特别强调抗Rorl自身抗体。(4)开发Project中生成的E mu-TCL 1小鼠模型 1评价旨在最大化Ad-CD 154基因治疗（单独或联合）活性的参数 与抗白血病药物或基于免疫的治疗策略组合。

Patients treated with autologous CLL cells transduced with an adenovirus encoding murine CD154 (Ad- CD154) experienced acute declines in leukemia cell counts and lymph node size and long-term desirable effects, including increased numbers of leukemia-reactive T cells, generation of anti-CLL antibodies, and apparent slowing or halting of disease progression. Treated patients had changes in bystander nontransfected CLL cells, reflecting activation by CD154 in vivo. We found that CD154-activation induced a programmed series of events in CLL cells, which initially were transiently protected and then sensitized to apoptosis induced by ligation of induced extrinsic death receptors (e.g. CD95 and DR5), cytotoxic drugs, or novel agents that inhibit the inhibitor of apoptosis (IAP) proteins (see project 2). Sensitivity to these agents appears associated with the capacity of CD154-activation to induce expression of the pro-apoptotic molecule Bid through a p53-independent, c-Abl-dependent mechanism, which apparently involves the alpha isoform of p73. This raises the exciting prospect that Ad-CD154 gene therapy may circumvent the dependency on p53 of anti-leukemia drugs, while inducing anti-leukemia cellular and antibody immune responses. The latter already have allowed us to identify a novel CLL-associated antigen, Ror1, which potentially could be used in assays to monitor the activity of Ad-CD154 gene therapy or in vaccines for immune therapy of this disease. For future clinical studies, we developed a "humanized" CD154 (designated ISF35) that can be stably expressed at high-levels on the CLL plasma membrane. This project has the following specific aims: (1) Continue ongoing analyses of the mechanism(s) responsible for the acute fall in leukemia-cell counts observed in patients treated with autologous Ad-CD154-transduced, or ISF35-transduced, CLL cells; (2) Examine cellular immune responses against a newly-identified CLL-associated antigen, Ror1; (3) Evaluate the specificity and biologic activity of anti-leukemia antibodies induced by Ad-CD154 gene therapy, with special emphasis on anti-Rorl autoantibodies. (4) Develop the E mu-TCL1 mouse model generated in Project 1 to evaluate parameters intended to maximize the activity of Ad-CD154 gene therapy, either alone or in combination with anti-leukemia drugs or immune-based treatment strategies.