TCR TRANSDUCED CD4 T CELLS FOR ADOPTIVE IMMUNOTHERAPY

在我们最初的研究中，我们首先描述了通过逆转录病毒转导带有MHC I类限制性TCR基因的正常T细胞产生MHC I类限制性CD4+ T细胞的能力。我们随后发现，如果TCR具有足够的亲和力，所产生的MHC 1类限制性CD4+ T细胞可以识别肿瘤细胞表达的生理水平的抗原。因此，这些新型T细胞可以通过在肿瘤病变中启动宿主免疫反应来增强抗肿瘤免疫反应。它们还能促进过继转移的CD8+ T细胞的持久性和功能。然而，对于体内TCR转导CD4+ T细胞的生物学特性及其对体内体外CD8+ T细胞的影响尚不清楚。我们有初步的数据显示，这种新种群实际上抑制了CD8+ T细胞的启动，这与它们预期的功能相反。该项目的目标是更好地了解MHC 1类限制性CD4+ T细胞在抗肿瘤免疫中的作用。我们的中心假设是MHC I类受限，TCR转导的CD4+ T细胞可以通过CD8+ T细胞增强抗肿瘤免疫反应。我们预测这将通过诱导它们成为能够在体外将DC许可给初始CD8+ T细胞的强效Th细胞来实现。我们进一步预测MHC I类受限，TCR转导的CD4+ T细胞可以在体内促进TCR转导的CD8+ T细胞的持久性和功能。这些假设/预测将通过小鼠和人类CD4+ T细胞转导表达TIL 13831 TCR的组合进行测试。这些TCR转导的CD4+ T细胞识别HLA-A2呈递的酪氨酸酶368-376表位，将与正常小鼠或人类的CD4+ T细胞分泌细胞因子的能力进行比较，允许DC启动/激活初始和TCR转导的CD8+ T细胞，并介导体内肿瘤消退。

In our original studies, we first described the ability to generate MHC class I restricted CD4+ T cells by retrovirally transduced normal T cells with MHC class I restricted TCR genes. We subsequently showed that if the TCR had sufficient affinity, the resulting MHC class 1 restricted CD4+ T cells could recognize physiologic levels of antigen expressed by tumor cells. Therefore, these novels T cells could augment the anti-tumor immune response by helping to prime the host immune response in tumor lesions. They could also promote the persistence and function of adoptively transferred CD8+ T cells. However, nothing is known about the biology of TCR transduced CD4+ T cells in vivo and their impact on the CD8+ T cells in vitro or in vivo. We have preliminary data that shows this novel population actually inhibits CD8+ T cell priming which would be contrary to their desired function. The goal of this project is to acquire a better understanding ofthe role of MHC class 1 restricted CD4+ T cells in anti-tumor immunity. Our central hypothesis is that MHC class I restricted, TCR transduced CD4+ T cells can be made to augment the antitumor immune response by CD8+ T cells. We predict this will occur by inducing them to become potent Th cells capable of licensing DC to prime CD8+ T cells in vitro. We further predict that MHC class I restricted, TCR transduced CD4+ T cells can be made promote the persistence and function of TCR transduced CD8+ T cells in vivo. These hypotheses/predictions will be tested using a combination of mouse and human CD4+ T cells transduced to express the TIL 13831 TCR. These TCR transduced CD4+ T cells, which recognize the tyrosinase:368-376 epitope presented by HLA-A2, will be compared to their normal mouse or human counterparts for their ability secrete cytokines, license DC to prime/activate naive and TCR transduced CD8+ T cells, and mediate tumor regression in vivo.