Mechanisms of Vaccine Immunity against Coccidioidomycosis

摘要 我们将开发一种活体减毒的∆cps1(空)球虫Posadasii株来接种球孢子虫病疫苗，以及 阐明肺上皮细胞协调诱导持久T细胞免疫抵抗感染的机制。 宿主对吸入病原体的抵抗力被认为存在于肺组织驻留记忆(TRM)细胞和粘膜中 免疫，但鲜为人知的真菌疫苗是否或如何创造常驻T细胞免疫在肺粘膜。 我们的初步数据显示，粘膜疫苗可以诱导保护性的球虫内切葡聚糖酶2(C-Eng2)特异性 疫苗接种后，肺脏上皮细胞需要动员髓系细胞和淋巴样细胞。 这一基质和造血细胞对接种疫苗的反应网络尚未被研究，这是本项目的主题。 为了确定在肺粘膜引起保护性免疫所需的细胞和分子机制，我们创造了 创新工具：i)一种新的保护性抗原C-Eng2和多肽：MHCII四聚体，用于跟踪和计数保护性CD4+ 小鼠C57BL6中的T细胞；II)上皮细胞及其产物能够有条件地缺失的转基因小鼠 分析它们在免疫生成中的作用；以及(Iii)分离和培养人肺上皮细胞的新系统 将结果从老鼠转化为人类。我们推测，∆cps1疫苗孢子的粘膜递送促进了肿瘤的发育。 保护的TRM细胞的开放和肺上皮协调持久的保护性免疫。我们的目标是： 1.探讨肺内天然免疫细胞对疫苗识别和应答的初始阶段。我们将检验这一假设 肺上皮及其产物调节识别和抑制生长的先天免疫细胞的动员 ∆cps1疫苗孢子，并捕获它们，将它们的多肽呈现给引流淋巴结中的幼稚T细胞。 2.阐明肺上皮细胞对∆cps1疫苗先天免疫应答的细胞内信号转导途径。 我们将检验这一假说，即肺上皮细胞内的钙和钙调神经磷酸酶依赖的信号事件有助于 球虫对宿主炎症反应的Dectin-1依赖性。并在老鼠和人类身上接种疫苗。 3.明确在肺粘膜接种疫苗引起的持久T细胞免疫的机制。我们将检验这一假设 肺上皮形成保护性C-Eng2特异性CD4+的表型、功能和转录特征 ∆-cps-1疫苗孢子经口给药后的T细胞。 我们的工作将开发一种高效的球孢子菌病疫苗来阐明肺免疫的机制。 在粘膜给药后。目前对疫苗诱导保护性免疫的机制知之甚少。 肺粘膜，以及肺上皮如何调节疫苗孢子的早期识别和抑制以及疫苗的产生 保护性T细胞。球孢子菌病疫苗接种是一个重要的公共卫生问题，也是一个重大的未得到满足的医学问题。 需要。我们期待着对疫苗诱导粘膜耐药的机制基础的新见解，以及 疫苗耐药性的免疫相关性将推动球孢子菌病预防措施的发展。

ABSTRACT We will exploit a live attenuated ∆cps1 (null) strain of Coccidiodes posadasii to vaccinate against coccidiodomycosis, and elucidate the mechanics by which the lung epithelium coordinates the induction of durable T cell immunity against infection. Host resistance against inhaled pathogens is thought to reside with lung tissue-resident memory (TRM) cells and mucosal immunity, but little is known about whether or how a fungal vaccine creates resident T cell immunity at the lung mucosa. Our preliminary data reveal that mucosal vaccination elicits protective, Coccidioides endoglucanase 2 (C-Eng2) specific CD4+ T cells and that the lung epithelium is required to mobilize myeloid and lymphoid cells in response to the vaccine. This network of stromal and hematopoetic cell responses to vaccination has not been studied and is the subject of this project. To define cellular and molecular mechanisms required to elicit protective immunity at the lung mucosa, we have created innovative tools: i) a novel protective antigen C-Eng2 and peptide:MHCII tetramers to track and enumerate protective CD4+ T cells in C57BL6 in mice; ii) transgenic mice in which epithelial cells and their products are able to be conditionally deleted to dissect their role in generation of immunity; and (iii) a novel system of isolating and culturing human lung epithelium to translate results from mice to humans. We hypothesize that the mucosal delivery of ∆cps1 vaccine spores promotes devel- opment of protective TRM cells and that the lung epithelium coordinates durable protective immunity. We aim to: 1. Probe the intial stages of vaccine recognition and response by innate immune cells in lung. We will test the hypothesis that lung epithelium and their products regulate mobilization of innate immune cells that recognize and restrain growth of ∆cps1 vaccine spores, and also captures them to present their peptides to naïve T cells in the draining lymph nodes. 2. Elucidate intracellular signaling pathway during lung epithelial cell regulation of innate response to ∆cps1 vaccine. We will test the hypothesis that Ca++ and calcineurin dependent signaling events within the lung epithelium contribute to the dectin-1 dependency of host inflammatory responses to Coccidiodes spp. and vaccine in both mice and humans. 3. Define mechanisms of durable T cell immunity elicited by vaccination at the lung mucosa. We will test the hypothesis that lung epithelium shapes the phenotypic, functional and transcriptomic features of protective C-Eng2-specific CD4+ T cells following intransal administration ∆cps1 vaccine spores . Our work will exploit a highly effective vaccine against coccidioidomycosis to elucidate the mechanisms of lung immunity following mucosal administration. Little is known about the mechanisms of vaccine-induced protective immunity at the lung mucosa, and how the lung epithelium regulates the early recognition and restraint of vaccine spores and generation of protective T cells. Vaccination against coccidioidomycosis, an important public health problem, is a major unmet medical need. We anticipate that new insight into the mechanistic underpinnings of vaccine induced mucosal resistance, and the immune correlates of vaccine resistance will advance the development of preventive measures against coccidioidomycosis.