Defining the role of T-bet and Eomes in CD8 T cell tolerance versus immunity

定义 T-bet 和 Eomes 在 CD8 T 细胞耐受与免疫中的作用

• 批准号: 8693610
• 负责人: Stephanie Renae Jackson
• 金额: $ 4.77万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8693610
• 关键词: Adoptive Immunotherapy; Boxing; CD8B1 gene; CXCR3 gene; Clinical; Data; Disease; Environment; Fostering; Functional disorder; Gene Targeting; Genes; Genetic; Human; Immune response; Immune system; Immunity; Immunotherapy; In complete remission; Individual; Interferons; Knock-out; Knockout Mice; Lead; Listeria monocytogenes; Malignant - descriptor; Malignant Neoplasms; Mediating; Molecular; Molecular Target; Mus; Pathway interactions; Patients; Peripheral; Phenotype; Process; Production; Proteins; Role; Signal Transduction; Specificity; T cell differentiation; T cell regulation; T cell response; T cell therapy; T-Lymphocyte; T-bet protein; Time; Transcript; Transgenic Mice; Translations; Treatment Efficacy; Tumor Immunity; Vaccinated; Vaccination; Vaccines; autoreactive T cell; base; cancer cell; cancer immunotherapy; cancer therapy; cell type; cytokine; design; granzyme B; improved; in vivo; insight; leukemia; mouse model; perforin; peripheral tolerance; pre-clinical; prevent; public health relevance; research study; response; tool; trafficking; transcription factor; tumor; tumor progression

DESCRIPTION (provided by applicant): Adoptive T cell immunotherapy strives to harness the power and specificity of the immune system to cure cancer in human patients. This approach relies on the transfer of tumor-reactive CD8+ T cells into patients, but the anticipated clinical benefits have not been achieved due to the induction of T cell tolerance. Peripheral tolerance is a multifaceted process, characterized by compromised proliferation, impaired effector function, and deletion of autoreactive T cells. Efforts to overcome tolerance for improved immunotherapy have been hampered because the T cell intrinsic pathways that regulate whether tumor/self-reactive T cells are directed toward tolerance versus immunity have yet to be defined. Using a pre-clinical mouse model of T cell tolerance, we found that Listeria monocytogenes vaccination at the time of adoptive CD8+ T cell transfer prevents tolerance. Specifically, transferred tumor/self-reactive T cells acquired effector function, displayed increased in vivo persistence, and provided a survival benefit to tumor-bearing hosts receiving adoptive T cell immunotherapy. T cell rescue corresponded with expression of the transcription factors T-bet and Eomesodermin (Eomes), both of which failed to be induced under tolerizing conditions in the absence of vaccination. T-bet and Eomes have a defined role in dictating type-I (IFN-? mediated) immune responses. Aside from their role in promoting IFN-? expression, these transcription factors also influence expression of several other genes that encode effector molecules important for CD8+ T cell trafficking or control of infected or malignant cells, includin CXCR3, granzyme B, perforin, and FasL. Our data suggest that T-bet and Eomes may be fundamental to the decision of T cell immunity versus tolerance, and as such, we propose to decipher the role of these transcription factors in this important fate decision. To this end, we wll utilize mice with genetic modulation of T-bet and Eomes in peripheral tumor/self-reactive CD8+ T cells as tools to dissect the role of these transcription factors during induction of T cell tolerance and under conditions of vaccine-mediated T cell rescue. Specific Aim I will focus on the individual and collective contributions of T-bet and Eomes to vaccine-mediated rescue of peripheral CD8+ T cell tolerance. We will evaluate effector cytokine production, T cell persistence, and transcript and protein production for T-bet or Eomes target genes in T cells rendered unable to upregulate T-bet, Eomes, or both; or T cells with constitutive T-bet expression. Specific Aim II will then focus on characterizing the specific contributions of these molecules to providing anti-tumor immunity within a tolerizing environment. Survival of tumor-bearing hosts will be used as a readout for therapeutic efficacy of adoptive immunotherapy with CD8+ T cells rendered genetically unable to express T-bet, Eomes or both; or T cells with constitutive T-bet expression. Identifying the contributions of T-bet and Eomes to T cell dysfunction during CD8+ T cell tolerance and adoptive immunotherapy may lead to improvements in T cell-based therapies for patients with cancer.

描述（由申请人提供）：产物T细胞免疫疗法旨在利用免疫系统治愈人类患者的癌症的功能和特异性。这种方法依赖于将肿瘤反应性CD8+ T细胞转移到患者中，但是由于T细胞耐受性的诱导，预期的临床益处尚未实现。外围耐受性是一个多方面的过程，其特征在于增殖受损，效应子功能受损和自动反应性T细胞的缺失。克服对改善免疫疗法的耐受性的努力受到阻碍，因为调节肿瘤/自我反应性T细胞是否针对耐受性与免疫力的T细胞内在途径尚未定义。使用T细胞耐受性的临床前小鼠模型，我们发现在收养CD8+ T细胞转移时，单核细胞增生李斯特菌疫苗接种可阻止耐受性。具体而言，转移的肿瘤/自我反应性T细胞获得了效应子功能，显示体内持久性增加，并为接受过继的T细胞免疫疗法的承重宿主提供了生存益处。 T细胞救援与转录因子T-bet和Eomesodermin（EOMES）的表达相对应，在没有疫苗接种的情况下，这两者都无法在耐受条件下诱导。 T-BET和EOME在决定I型（IFN-？介导的）免疫反应中具有明确的作用。除了他们在促进IFN-的角色外？表达，这些转录因子还会影响其他几种对CD8+ T细胞运输或对感染或恶性细胞的控制重要的效应分子的表达，包括CXCR3，Granzyme B，Perforin和Fasl。我们的数据表明，T-BET和EOMES可能是T细胞免疫与公差的决策基础，因此，我们建议破译这些转录因子在这一重要命运决策中的作用。为此，我们将T-BET和EOME的遗传调节的小鼠在周围肿瘤/自反应性CD8+ T细胞中用作剖析这些转录因子在诱导T细胞耐受性和疫苗介导的T细胞救援的条件下的作用的工具。具体目的我将重点关注T-BET和EOMES对疫苗介导的外围CD8+ T细胞耐受性的营救的个体和集体贡献。我们将评估效应子细胞因子的产生，T细胞的持久性以及T-BET或EOMES靶基因的T细胞中无法上调T-BET，EOMES或两者兼而有之的T-BET或EOMES靶基因的转录和蛋白质的产生。或具有组成型T-bet表达的T细胞。然后，特定的目标II将集中于表征这些分子在耐受性环境中提供抗肿瘤免疫的特定贡献。含肿瘤宿主的存活率将用作用CD8+ T细胞产生的遗传学上无法表达T-bet，eomes或两者两者的CD8+ T细胞的治疗功效的读数。或具有组成型T-bet表达的T细胞。在CD8+ T细胞耐受性和收养免疫疗法期间，鉴定T-Bet和Eomes对T细胞功能障碍的贡献可能会改善基于T细胞的癌症患者的T细胞疗法。