ADOPTIVE IMMUNOTHERAPY UTILIZING T CELLS INDUCED BY GENE MODIFIED HUMAN CANCERS

利用基因修饰人类癌症诱导的 T 细胞进行过继免疫治疗

• 批准号: 5209287
• 负责人: ALFRED E CHANG
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5209287
• 关键词: CD28 molecule; CD3 molecule; MHC class I antigen; clinical trials; cytotoxic T lymphocyte; genetic transduction; host neoplasm interaction; human subject; human therapy evaluation; immunosuppression; interleukin 2; laboratory mouse; monoclonal antibody; neoplasm /cancer genetics; neoplasm /cancer immunotherapy; neoplasm /cancer vaccine; neoplastic cell; tissue /cell culture; transfection; tumor necrosis factor alpha

Although it has been well established experimentally that the transfer of sensitized T lymphocytes can mediate potent antitumor effects, extrapolating the principles of adoptive immunotherapy obtained from animal studies to clinical therapy will require the development of innovative techniques to isolate and propagate antitumor effector T cells from cancer patients. Toward this end, we have established culture methods whereby cells from tumor-draining or tumor primed lymph nodes (LN) can be sensitized to acquire therapeutic efficacy. Since these cells do not demonstrate overt antitumor reactivity before culture, they are functionally referred to as "pre-effector" cells. One method involves the in vitro sensitization (IVS) of pre-effector cells with tumor cells in the presence of IL-2. Another method involves the sequential activation of pre-effector LN cells with anti-CD3 mAb followed by expansion in low concentrations of IL-2. In preliminary clinical studies, we have demonstrated the feasibility of inducing vaccine primed LN cells by the in vivo inoculation of autologous tumor cells admixed with BCG followed by in vitro activation by the methods described. DTH responsiveness to autologous tumor was conferred by the adoptive transfer of activated cells; and tumor regression was evident in a select group of patients. Major obstacles which confront the clinical applications of adoptive immunotherapy include the relatively poor immunogeneicity of human cancers and the potential problems of tumor-induced immunosuppression. Recent observations in animal studies indicate the tumors can be genetically altered to enhance the host immune response against native or parental tumor antigens. The transfection of tumor cells with specific cytokine genes have resulted in T cell mediated rejection of these tumors by their syngeneic hosts. In a different approach, we have found that the transfection of murine tumor cells by an allogeneic MHC class I gene will upregulate pre-effector cell sensitization against the poorly immunogenic B16BL6 melanoma; abrogate tumor-induced suppression of pre-effector cell induction; and can mediate regression of large established tumors by a novel in vivo transfection technique. These observations provide an impetus for the design of a clinical protocol examining the efficacy of gene-modified tumors utilized as a vaccine to induce pre-effector LN cells in cancer patients for subsequent adoptive immunotherapy. The specific aims of the current proposal are: (1) TO conduct a clinical trial of adoptive T cell immunotherapy with anti- CD3/IL-2 activated LN cells induced by the inoculation of gene-modified tumor cells, (2) To assess the toxicity, antitumor efficacy, in vivo immunological reactivity of patients, and in vivo homing of the activated cells, and (3) To characterize the in vitro antigen specificity of the sensitized T cells, and investigate methods of optimal sensitization of T cells utilizing cytokines and/or anti-CD28 mAb.

尽管实验证明， 致敏的T淋巴细胞可以介导有效的抗肿瘤作用， 从以下获得的过继免疫疗法的原理外推 从动物研究到临床治疗需要开发 分离和增殖抗肿瘤效应T创新技术 癌症患者的细胞。为此，我们建立了文化 来自肿瘤引流或肿瘤引发的淋巴结的细胞 (LN)可被敏化以获得治疗功效。由于这些 细胞在培养前没有表现出明显的抗肿瘤反应性， 在功能上称为“前效应”细胞。一种方法 涉及前效应细胞的体外致敏（IVS）， IL-2存在下的肿瘤细胞。另一种方法涉及 用抗-CD 3 mAb顺序激活前效应LN细胞 随后在低浓度IL-2中扩增。初步 临床研究，我们已经证明了诱导的可行性 通过体内接种自体肿瘤的疫苗致敏的LN细胞 细胞与BCG混合，然后通过以下方法体外活化 介绍了DTH对自体肿瘤的反应性由 活化细胞的过继转移;肿瘤消退明显 在一组特定的病人中。 过继免疫疗法临床应用面临的主要障碍 免疫疗法包括人类相对较差的免疫原性 癌症和肿瘤诱导的免疫抑制的潜在问题。 最近在动物研究中的观察表明，肿瘤可以是 基因改变以增强宿主对天然抗体的免疫应答， 或亲本肿瘤抗原。肿瘤细胞的转染 特异性细胞因子基因导致T细胞介导的排斥， 这些肿瘤是由其同基因宿主引起的。在另一种方法中，我们 发现用同种异体MHC转染小鼠肿瘤细胞 I类基因将上调前效应细胞对 免疫原性差的B16 BL 6黑色素瘤;消除肿瘤诱导的抑制 前效应细胞诱导;并可介导大的 通过新的体内转染技术建立肿瘤。这些 观察结果为临床方案的设计提供了动力 研究基因修饰肿瘤作为疫苗的有效性， 在癌症患者中诱导前效应LN细胞用于随后过继 免疫疗法目前建议的具体目标是：（1） 进行过继性T细胞免疫治疗的临床试验， 基因修饰的IL-2诱导LN细胞活化 肿瘤细胞，（2）评估体内毒性、抗肿瘤功效 患者的免疫反应性，和体内归巢的 活化的细胞，和（3）表征体外抗原 致敏T细胞的特异性，并研究方法， 利用细胞因子和/或抗CD 28的T细胞的最佳致敏 mAb.