EXPANSION OF ANTI-TUMOR T CELLS FROM TUMOR-BEARING HOSTS

来自荷瘤宿主的抗肿瘤 T 细胞的扩增

• 批准号: 3192027
• 负责人: HARRY D BEAR
• 金额: $ 18.94万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-15 至 1994-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3192027
• 关键词: CD3 molecule; breast neoplasms; calcium flux; cellular oncology; cimetidine; clone cells; cyclophosphamide; cytotoxic T lymphocyte; disease /disorder model; genetic markers; host neoplasm interaction; human subject; human tissue; immunomodulators; interleukin 2; laboratory mouse; leukocyte activation /transformation; neoplasm /cancer immunology; neoplasm /cancer immunotherapy; neoplasm /cancer surgery; neoplasm /cancer transplantation; phorbols; protein kinase C; suppressor T lymphocyte; tissue /cell culture; transforming growth factors; tumor antigens; tumor necrosis factor alpha

For effective adoptive immunotherapy of cancer, tumor-specific T lymphocytes must be obtained from tumor-bearing patients, activated in vitro and expanded to large numbers, but tumor-induced suppressor cells and the unavailability of tumor antigen may critically limit the expansion of tumor-reactive T cells. In murine tumor models, we have found that T cells from tumor-draining lymph nodes (DLN) are more readily expanded than spleen cells from the same hosts, and that these cells have therapeutic activity in vivo. Low dose cyclophosphamide in vivo and an analogue (mafosfamide) used in vitro can inhibit the ability of suppressor cells to limit cytotoxic T lymphocyte (CTL) growth, without adversely affecting CTL activation. CTL from syngeneic murine P815 tumor-bearing host (TBH) spleens, which would otherwise not grow, could be expanded after in vitro treatment with mafosfamide. Using pharmacologic manipulation of the pathways of T cell signal transduction, via activation of protein kinase C and increasing intracellular Ca++, in place of antigen we could expand T cells which were cytotoxic in vitro and therapeutic in vivo. We have also derived tumor-specific CTL clones from TBH mice which not only depend upon both antigen and exogenous interleukin 2 (IL-2) for growth, but also can attain a "resting" state similar to memory CTL from TBHS. From this resting state, we can distinctly separate the antigen-dependent and IL-2- dependent stages of T cell activation. Thus, we have a unique opportunity to examine in detail the regulation of T cell activation in a tumor- specific, non-transformed and regulable CTL clone. These cloned CTL also have therapeutic activity in vivo and will serve as a model on which to base rational manipulations of lymphocytes from TBH which will maximize T cell growth and preserve anti-tumor activity. Moreover, the above methods of suppressor cell inhibition and pharmacologic T cell activation are applicable to DLN from human patients with breast cancer, but the responsiveness of cells from individual nodes is highly variable. This project aims to: (1) Determine how different methods of T cell activation can be used to expand CTL clones and to delineate the detailed cellular and molecular mechanisms by which regulatory cytokines (IL-2, IL-4 and TGF-Beta) affect the growth of cloned CTL; (2) Determine whether similar methods, combined with inhibition of Ts, function by mafosfamide, can be applied to the expansion of bulk cultures of T cells obtained directly from TBH mice, so that the yield of cells is maximized and anti- tumor activity is maintained; (3) Test the T cells obtained by different, strategies of expansion for therapeutic efficacy in vivo; and (4) Determine how to identify, isolate and expand T cells with potential anti- tumor activity from the DLN of human breast cancer patients, using similar strategies, including mafosfamide treatment and pharmacologic activation of T cells. The proposed studies should lead to a clearer. understanding of tumor-specific CTL activation, regulation and growth and to more rational and more effective strategies for clinical adoptive immunotherapy of cancer.

对于癌症的有效过继免疫治疗，肿瘤特异性 T 淋巴细胞必须从携带肿瘤的患者中获得，并在 体外并大量扩增，但肿瘤诱导的抑制细胞 肿瘤抗原的缺乏可能会严重限制扩张 肿瘤反应性 T 细胞。 在小鼠肿瘤模型中，我们发现 T 来自肿瘤引流淋巴结（DLN）的细胞比来自肿瘤引流淋巴结（DLN）的细胞更容易扩增 来自同一宿主的脾细胞，并且这些细胞具有治疗作用 体内活性。 体内低剂量环磷酰胺及其类似物 （mafosfamide）在体外使用可以抑制抑制细胞的能力 限制细胞毒性 T 淋巴细胞 (CTL) 的生长，而不会对 CTL 产生不利影响 激活。 来自同基因小鼠 P815 荷瘤宿主 (TBH) 的 CTL 脾脏原本不会生长，但在体外培养后可以扩大 用mafosfamide治疗。 利用药理操作 通过激活蛋白激酶，T 细胞信号转导途径 C 和增加细胞内 Ca++，我们可以扩展代替抗原 T细胞在体外具有细胞毒性，在体内具有治疗作用。 我们有 还从 TBH 小鼠中衍生出肿瘤特异性 CTL 克隆，这不仅取决于 依靠抗原和外源白细胞介素 2 (IL-2) 进行生长，而且 可以达到类似于 TBHS 的记忆 CTL 的“休息”状态。 从这里 静息状态下，我们可以清楚地区分抗原依赖性和IL-2- T 细胞激活的依赖阶段。 因此，我们有一个独特的机会 详细检查肿瘤中 T 细胞激活的调节 特异性、非转化且可调节的 CTL 克隆。 这些克隆的 CTL 还 具有体内治疗活性，并将作为模型 对来自 TBH 的淋巴细胞进行基本合理的操作，这将使 T 最大化 细胞生长并保持抗肿瘤活性。 另外，上述方法 抑制细胞抑制和药理 T 细胞激活的作用 适用于人类乳腺癌患者的 DLN，但 来自各个节点的细胞的响应性变化很大。 该项目旨在： (1) 确定 T 细胞的不同方法如何 激活可用于扩展 CTL 克隆并描绘详细的 调节细胞因子（IL-2、 IL-4和TGF-Beta)影响克隆CTL的生长； (2) 判断是否 类似的方法，结合 mafosfamide 抑制 Ts 的功能， 可应用于获得的 T 细胞批量培养物的扩增 直接来自TBH小鼠，使细胞产量最大化并抗- 维持肿瘤活性； (3) 测试不同获得的T细胞， 扩大体内治疗功效的策略；和（4） 确定如何识别、分离和扩增具有潜在抗-T细胞 使用类似的方法从人类乳腺癌患者的 DLN 中获取肿瘤活性 策略，包括马磷酰胺治疗和药理激活 T 细胞。 建议的研究应该会带来更清晰的结果。理解 肿瘤特异性 CTL 激活、调节和生长以及更多 临床过继免疫治疗的合理且更有效的策略 癌症。