TCR DNA IMMUNIZATION FOR 2B411 T CELL LYMPHOMA

皮肤T细胞淋巴瘤（CTCL）患者的恶性细胞 是克隆的，并且共享不同的T细胞受体（TCR）。的前提 这个项目是，抗肿瘤免疫反应可以产生， 接种编码TCR基因的TCR-DNA表达质粒 T细胞淋巴瘤中恶性克隆特异性片段。 这 将在已建立的T细胞淋巴瘤鼠模型中进行测试，其中 将2B4.11 T细胞杂交瘤细胞皮下注射到 同源小鼠导致肿瘤生长和存活率降低。 五 一年的指导计划，提出了在小鼠中研究的安全性，有效性， 以及各种旨在增强T细胞DNA免疫疗法的策略 淋巴瘤 R. Tigelaar（细胞免疫学专家）将担任 sponsor和A. Hayday（分子免疫学专家）和J. Brandsma （DNA免疫专家）将担任共同赞助者。 用编码α和β-受体部分的TCR-DNA质粒接种 将测定2B 4 TCR的β链诱导免疫应答的能力。 特异性抗肿瘤（抗-2B4.11）免疫应答，通过 体外增殖和细胞毒性测定。接种的小鼠也将 在体内测试它们抵抗2B4.11肿瘤攻击的能力。 CD 4+和CD 8+细胞对这些反应的相对贡献 将使用体外耗尽的细胞群来确定，以及 体内消耗的小鼠。 直接比较将检验以下方面的相关性： 接种技术（皮内vs.肌内vs.基因枪）和 接种次数（单次与多次）对TCR-DNA 质粒诱导肿瘤免疫和保护，如在 以上分析。 一旦免疫的最佳条件在这个 模型已经确定TCR-DNA质粒将被测试其 治疗先前用2B4.11杂交瘤肿瘤攻击的小鼠的能力。 各种旨在增强免疫力的策略 将检查TCR-DNA质粒免疫的潜力，包括： (1)与含有CpG基序的寡核苷酸共接种，（2）共- 或预先注射细胞因子表达质粒（即GM-CSF、IL-12）， (3)用嵌合TCR/细胞运输蛋白（即， 泛素，LAMP-1）质粒，和（4）用树突状细胞接种 用TCR-DNA质粒体外转化。 成功瞄准 通过TCR-DNA免疫的致病性T细胞群可能导致 CTCL和其他系统性白血病/淋巴瘤的潜在治疗 自身免疫过程，如多发性硬化症和移植 并发症，如GVHD和器官移植排斥。

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.