ADOPTIVE IMMUNOTHERAPY UTILIZING T CELLS INDUCED BY GENE MODIFIED HUMAN CANCERS

尽管实验已经很好地证实了这种转移 致敏的T淋巴细胞可以介导强大的抗肿瘤作用， 推论过继免疫疗法的原理 动物研究到临床治疗将需要发展 分离和扩增抗肿瘤效应T细胞的创新技术 来自癌症患者的细胞。为此，我们建立了文化 从肿瘤引流或肿瘤引流的淋巴结中提取细胞的方法 (Ln)可被致敏以获得治疗效果。因为这些 细胞在培养前没有表现出明显的抗肿瘤反应，它们 在功能上被称为“前效应器”细胞。一种方法 涉及前效应细胞的体外增敏(IVS) 肿瘤细胞中存在IL-2。另一种方法涉及 抗CD3单抗序贯激活前效应层粘连细胞 然后在低浓度的IL-2中扩张。在预赛中 临床研究，我们已经证明了诱导的可行性 自体肿瘤体内接种疫苗激活的LN细胞 细胞与卡介苗混合后通过体外激活的方法 描述。DTH对自体肿瘤的反应性是由 过继转移活化细胞；肿瘤明显消退 在精选的一组病人中。 领养在临床应用中面临的主要障碍 免疫治疗包括人类相对较差的免疫原性 癌症和肿瘤引起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 经过基因改造以增强宿主对天然免疫的免疫反应 或亲代肿瘤抗原。基因转导肿瘤细胞的实验研究 特异性细胞因子基因导致T细胞介导的排斥反应 这些肿瘤是由它们的同基因宿主造成的。在不同的方法中，我们有 发现同种异体MHC对小鼠肿瘤细胞的转染 I类基因将上调前效应激反应细胞对 免疫原性差的B16BL6黑色素瘤；肿瘤诱导的抑制 前效应细胞的诱导；并可以介导大鼠的退变 通过一种新的体内转基因技术建立肿瘤。这些 观察结果为临床方案的设计提供了动力 检测转基因肿瘤作为疫苗用于治疗的效果 在癌症患者体内诱导前效应层粘连细胞用于后续传代 免疫疗法。目前建议的具体目标是：(1) 进行抗人T细胞过继免疫治疗的临床试验 转基因疫苗诱导CD3/IL-2激活的LN细胞 肿瘤细胞，(2)评估毒性，体内抗肿瘤效果 患者的免疫反应性和体内归巢 激活的细胞，以及(3)鉴定体外抗原 致敏T细胞的特异性，并探讨致敏T细胞 细胞因子和/或抗CD28对T细胞的最佳致敏作用 Mab.

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.