Regulation of vaccine-induced anti-fungal Th17 cells

项目摘要/摘要 传染病疫苗被誉为过去十年来公共卫生领域最伟大的成就 上个世纪。这一竞争更新将研究一种能引起细胞免疫反应的新型佐剂 需要更好的疫苗。缺乏适当的佐剂是开发一种安全和 有效预防真菌病原体感染的疫苗，这是医学和医疗领域尚未得到满足的需求 公共卫生。尽管严重真菌感染率不断上升，但目前还没有针对真菌的疫苗商业化。 可用。虽然目前针对传染病的疫苗优先诱导抗体的产生， 它们对宿主防御真菌的贡献是有限的。细胞免疫是解决疾病的关键 真菌感染。疫苗诱导的实验模型对真菌的耐药性需要Th17和Th1 产生IL-17和干扰素-γ并部分通过招募和激活吞噬细胞来介导保护的细胞 增强对真菌的杀灭作用。在上一个资金周期中，我们发现C型凝集素对真菌的识别 受体(CLR)Dectin-2是保护Th17和Th1细胞分化所必需的 真菌。通过生化提纯和质量光谱分析，鉴定了一种新的天然产物。 Dectin-2的真菌配基，糖蛋白Blastomyces Eng3(Bl-Eng3)。 在这项应用中，我们建议对作为疫苗佐剂的Bl-Eng3进行表征和功能测试 对抗真菌。我们假设Bl-Eng3激活树突状细胞，驱动抗原分化 (AG)特异性Th17和Th1细胞，从而促进对真菌的保护性细胞免疫 感染。我们还假设蛋白质骨架和糖链修饰是配体活性的原因。 而Bl-Eng3将单独或与其他CLR配体一起成为疫苗的有效佐剂 对抗真菌。我们提供了强有力的初步数据来支持我们的假设。使用Dectin-2报告细胞， 我们已经建立了一个体外筛选系统来分析蛋白质骨架和它的作用 糖基化Dectin-2配体活性和体内过继转移系统描述Dectin-2配体活性的影响 配基佐剂对单纯抗原特异性的CD4+T细胞向保护性Th17和Th1细胞分化的影响。 我们的方法提供了一个强大的补充策略，将确定糖蛋白的配体部分 目的1，评估抗原提呈细胞(APC)和抗原特异性T细胞成熟和启动的佐剂作用 在Aim 2中，并在实验模型中确定受体与其他CLR的协同作用和保护效果 我们的工作将提供所需的新见解，以促进 APC成熟及保护性Th17和Th1细胞分化。这些知识将提供基础 开发和设计新的疫苗策略，以对抗真菌和其他需要 用于宿主防御的细胞免疫。

Project Summary/Abstract Vaccines against infectious diseases have been hailed as the greatest achievement in public health over the last century. This competing renewal will investigate a novel adjuvant that elicits cellular immune responses needed for better vaccines. The lack of an appropriate adjuvant is one major barrier to developing a safe and effective vaccine against infections with fungal pathogens, which represents an unmet need in medicine and public health. Despite the rising rates of severe fungal infections, no vaccines against fungi are commercially available. While current vaccines against infectious diseases preferentially induce production of antibodies, their contribution to host defense against fungi is limited. Cellular immunity is essential for the resolution of fungal infections. Vaccine-induced resistance against fungi in experimental models requires Th17 and Th1 cells that produce IL-17 and IFN-γ and mediate protection in part by recruiting and activating phagocytes to augment killing of fungi. In the last funding cycle, we discovered that fungal recognition by the C-type lectin receptor (CLR) Dectin-2 is required for the differentiation of protective Th17 and Th1 cells against dimorphic fungi. Through biochemical purification and mass spectrophotometry analysis, we identified a novel bona-fide fungal ligand for Dectin-2, the glycoprotein Blastomyces Eng3 (Bl-Eng3). In this application, we propose to characterize and functionally test Bl-Eng3 as an adjuvant for vaccination against fungi. We hypothesize that Bl-Eng3 activates dendritic cells, drives differentiation of antigen (Ag)-specific Th17 and Th1 cells, and thereby promotes protective cellular immunity against fungal infection. We also posit that the protein backbone and glycan modifications account for ligand activity and that Bl-Eng3 will be an effective adjuvant alone or together with other CLR ligands for vaccines against fungi. We provide strong preliminary data to support our hypotheses. Using Dectin-2 reporter cells, we have established an in vitro screening system to dissect the contributions of the protein backbone and its glycosylation to Dectin-2 ligand activity and an in vivo adoptive transfer system to delineate the effects of the ligand adjuvant on the differentiation of naïve antigen-specific CD4+ T-cells into protective Th17 and Th1 cells. Our approach offers a powerful complimentary strategy that will identify the ligand moiety of the glycoprotein in aim 1, assess adjuvancy for the maturation and priming of Ag-presenting cells (APCs) and Ag-specific T cells in aim 2, and determine receptor collaboration with other CLRs and protective efficacy in experimental models of pulmonary and systemic fungal infections in aim 3. Our work will provide new insight needed to promote the maturation of APCs and differentiation of protective Th17 and Th1 cells. This knowledge will provide the basis for developing and designing new vaccine strategies against fungi, and other pathogenic microbes that require cellular immunity for host defense.