Long-term effects of developmental exposure to a mixture of thyroid disruptors associated with hydrofracking on T cell development and antimicrobial immunity

这项探索性研究提案解决了人们对内分泌干扰物威胁日益增长的担忧。 (EDC)非常规石油和天然气开采(UOG)过程中释放的水污染物对人类健康的影响。 具体地说，这些研究集中在4种UOG化学物质与甲状腺激素(TH)的混合物如何扰乱 活动影响免疫系统，到目前为止还没有系统的评估。我们已经开发出一种 可靠、灵敏和经济高效的两栖动物模型系统，特别适合于 研究早期接触UOG毒物的水性混合物对晚年免疫力的影响。 我们提出了一种比较生物学的方法来定义非洲爪哇胚胎后如何暴露于 具有TH活性的UOG水污染物的代表性混合物影响T细胞发育和 在晚年削弱抗菌素免疫反应。我们的理论基础是(1)几个EDC的混合 朝着一个荷尔蒙轴运动将使我们能够解决导致急性和持久的细胞机制 这种化学混合物对T细胞发育和功能的影响；(2)尽管TH轴影响 免疫系统方面，对干扰物对免疫的长期生物学影响仍知之甚少；以及 (3)非洲爪鼠是一种可靠的、特征广泛的、公认的围产儿模型 TH轴的内分泌学和生物医学研究，包括TH信号对TH轴的依赖 变态过程中成人型T细胞免疫分化的研究有利于实现的其他特征 我们的目标是向外发展蝌蚪，它们不受母体的影响，很容易接触到 在胚胎后发育期间暴露的风险，以及大规模暴露蝌蚪的成本效益 数字传给急救中心。来检验这样一种假设，即发育过程中暴露在TH干扰物混合物中 与UOG相关的改变T细胞发育，导致长期免疫紊乱 动态平衡和较差的抗菌免疫力，我们将确定：(1) UOG-EDCs的急性效应 蝌蚪T细胞分化与免疫动态平衡 ，通过定义蝌蚪暴露对 用流式细胞术、组织学和转录学研究动态平衡的淋巴细胞群； 长期影响 TH干扰物混合物对发育过程中免疫稳态和抗微生物免疫的影响 蜕变后的坚持 ，通过测量在稳定状态下的变化和对RANAV FV3的响应 (类似于痘病毒，如痘苗，一种强大的CD8 T细胞诱导剂)和海洋分枝杆菌(一种病原体 对于青蛙和人类，是一种强大的CD4T细胞诱导剂)；以及(3) 4个UOG-EDC中每一个的TH干扰活性 单独及其潜在的相加和协同作用。 来自感染两个人的创新趋同发现 非洲爪哇不同类型病原体(痘样FV3和分枝杆菌)的TH轴和免疫系统 与哺乳动物的显著保守将提供关于影响的新的和令人信服的证据 UOG-EDCs对人体健康相关免疫系统功能的影响。

This exploratory research proposal addresses growing concerns about the threats of endocrine disruptor (EDC) water pollutants released during unconventional oil and gas extraction (UOG) to human health. Specifically, these studies focus on how a mixture of 4 UOG chemicals with thyroid hormone (TH) disrupting activity affect the immune system, for which there is no systematic assessment to date. We have developed a reliable, sensitive and cost-effective model system in the amphibian Xenopus that is uniquely suited to investigate the effects of early life exposure to waterborne mixtures of UOG toxicants on immunity later in life. We propose a comparative biology approach in Xenopus to define how postembryonic exposure to a representative mixture of UOG water contaminants with TH activity impacts T cell development and weaken antimicrobial immune response later in life. Our rationale is that (1) a mixture of several EDCs acting toward one hormone axis will allow us to address cellular mechanism leading to acute and persistent effects of this mixture of chemicals on T cell development and function; (2) Although TH axis affects the immune system, little is still known about the long-term biological impacts of TH disruptors on immunity; and (3) Xenopus is a reliable, extensively characterized and widely recognized model of human perinatal endocrinology and biomedical research on the TH axis, including dependence of TH signaling for the differentiation of adult-type T cell immunity during metamorphosis. Other features advantageous for achieving our goal are the external development of tadpoles, which are free of maternal influence and easily accessible to exposure during postembryonic development, and the cost effectiveness of exposing tadpoles in large numbers to EDCs. To test the hypothesis that developmental exposure to a mixture of TH-disruptors associated with UOG alters T cell development, resulting in long-term perturbation of immune homeostasis and poorer antimicrobial immunity, we will determine: (1) acute effects of UOG-EDCs on tadpole T cell differentiation and immune homeostasis , by defining the impact of tadpole exposure on homeostatic lymphocyte populations using flow cytometry, histology and transcriptomics; (2) long-term effects on immune homeostasis and anti-microbial immunity of developmental exposure to a mixture of TH-disruptors persisting after metamorphosis , by measuring changes at steady state and in response to the ranavirus FV3 (similar to poxviruses such as vaccinia, a strong CD8 T cell inducer) and Mycobacterium marinum (a pathogen of frogs and humans, a strong CD4 T cell inducer); and (3) TH-disruptor activity of each of the 4 UOG-EDC alone and their potential additive and synergistic actions. Innovative convergent findings from infection with two different types of pathogens (pox-like FV3 and Mycobacteria) in Xenopus whose TH axis and immune system are remarkably conserved with those of mammals will provide novel and compelling evidence regarding effects of UOG-EDCs on the immune system function relevant to human health.