TCR DNA IMMUNIZATION FOR 2B411 T CELL LYMPHOMA

皮肤T细胞淋巴瘤(CTCL)患者的恶性细胞 都是克隆性的，共享一个独特的T细胞受体(TCR)。的前提是 这个项目是抗肿瘤免疫反应可以通过以下方式产生 用编码TCR基因的TCR-DNA表达载体接种 T细胞淋巴瘤中恶性克隆的特异片段。这 将在已建立的T细胞淋巴瘤小鼠模型中进行测试 皮下注射2B4.11 T细胞杂交瘤细胞 同基因小鼠会导致肿瘤生长和存活率下降。A五 年指导计划被提出在小鼠身上研究安全性、有效性、 以及旨在加强T细胞DNA免疫治疗的各种策略 淋巴瘤。R.Tigelaar(细胞免疫学专家)将担任 赞助商，A.Hayday(分子免疫学专家)，J.Brandsma (DNA免疫专家)将担任共同发起人。 用编码部分α和α基因的TCR-DNA质粒接种 2B4 TCR的β链将被检测其诱导能力 由In决定的特异性抗肿瘤(抗2B4.11)免疫反应 体外增殖和细胞毒性试验。接种的小鼠也会 在体内测试其抵抗2B4.11肿瘤挑战的能力。 CD4+和CD8+细胞在这些反应中的相对贡献 将使用体外耗尽的细胞群以及 体内耗尽的小鼠。直接比较将分析 接种技术(皮内注射、肌肉注射、基因枪)和 接种次数(单次接种与多次接种)对TCR-DNA能力的影响 用来诱导肿瘤免疫和保护作用的质粒 以上化验。一旦在这个地方免疫的最佳条件 模型已经确定TCR-DNA质粒将被测试其 治疗之前挑战2B4.11杂交瘤的小鼠的能力。 旨在加强免疫治疗的各种策略 将研究TCR-DNA质粒免疫的潜力，包括： (1)共接种含CpG基序的寡核苷酸，(2)共接种 或预先注射细胞因子表达质粒(即GM-CSF、IL-12)， (3)接种嵌合TCR/细胞运输蛋白(即 泛素、LAMP-1)质粒，以及(4)树突状细胞接种 用TCR-DNA质粒进行体外转化。成功锁定目标 经TCR-DNA免疫的T细胞致病群体可能导致 CTCL和其他白血病/淋巴瘤的潜在治疗方法 自身免疫过程，如多发性硬化症和移植 GVHD和器官移植排斥反应等并发症。

The malignant cells from a patient with cutaneous T cell lymphoma (CTCL) are clonal and share a distinct T cell receptor (TCR). The premise of this project is that anti-tumor immune responses can be generated by inoculation with TCR-DNA expression plasmids which encode the TCR gene segments specific for the malignant clone in a T cell lymphoma. This will be tested in an established murine model of T cell lymphoma where the subcutaneous injection of the 2B4.11 T cell hybridoma cells into syngeneic mice results in tumor growth and decreased survival. A five year mentored program is proposed to study in mice the safety, efficacy, and various strategies designed to enhance DNA immunotherapy for T cell lymphoma. R. Tigelaar (expert in cellular immunology) will serve as sponsor, and A. Hayday (expert in molecular immunology), and J. Brandsma (expert in DNA immunization) will serve as cosponsors. Inoculations with TCR-DNA plasmids encoding portions of the alpha and beta chain of the 2B4 TCR will be assayed for their ability to elicit specific anti-tumor (anti-2B4.11) immune responses, as determined by in vitro proliferation and cytotoxicity assays. Inoculated mice will also be tested in vivo for their ability to resist 2B4.11 tumor challenge. The relative contributions of CD4+ and CD8+ cells to these responses will be determined using in vitro depleted cell populations, as well as in vivo depleted mice. Direct comparisons will assay the relevance of inoculation technique (intradermal vs. intramuscular vs. gene gun) and number of inoculations (single vs. multiple) on the ability of TCR-DNA plasmids to induce tumor immunity and protection as measured in the above assays. Once the optimal conditions for immunization in this model have been determined TCR-DNA plasmids will be tested for their ability to treat mice previously challenged with 2B4.11 hybridoma tumor. A variety of strategies designed to enhance the immunotherapeutic potential of TCR-DNA plasmid immunization will be examined, including: (1) co-inoculation with oligonucleotides containing CpG motifs, (2) co- or pre-injection with cytokine expression plasmids (i.e. GM-CSF, IL-12), (3) inoculation with chimeric TCR/cell-trafficking protein (i.e. ubiquitin, LAMP-1) plasmids, and (4) inoculation with dendritic cells transformed in vitro with TCR-DNA plasmids. Successfully targeting pathogenic populations of T cells via TCR-DNA immunization may lead to potential treatments for CTCL and other leukemias/lymphomas, systemic autoimmune processes such as multiple sclerosis, and transplant complications such as GVHD and organ transplant rejection.