Assays for acceleration: from fit-for-purpose models to scalable assays of broad systemic and mucosal protection against all strains of Group A Streptococcus

项目总结/摘要 国际上重新出现了研制疫苗的势头，以预防由高度流行性感冒引起的感染。 适应的人类限制性细菌病原体A组链球菌（GAS）。然而，缺乏一个已知的 人对GAS感染的免疫保护与目前体外测定的局限性的相关性， 临床前模型阻碍了有前途的临床前候选疫苗的开发，并威胁到未来的发展。 对GAS疫苗发现和设计的投资。为了克服这一障碍，我们将进行一次跨- 学科合作研究计划，利用高度相关的人类样本，开发一种新的 人类离体组织模型，以发现广泛适用的潜在人类保护相关物， 为适合在临床疫苗试验中部署的实用免疫测定提供信息设计。而不是坚持 由于历史GAS免疫测定的限制，我们明确针对临床应用的首选特征， 支持疫苗开发的免疫原性测定，如实用性、准确性和广泛应用， 相关证候和菌株。 天然气只感染人类，因此我们将采取以人为本的方法，有目的地远离 这些动物模型不能充分代表人类免疫反应的相关复杂性。我们 将应用最先进的免疫学方法来研究来自平行研究的各种人类样本 项目，包括使用我们自己的GAS人类挑战模型评估疫苗的外部资助试验 预防咽炎，这是疫苗开发工作的主要目标适应症。首次出版 来自GAS咽炎人类挑战模型的发现表明，保护可能与强大的 早期粘膜和全身性Th 1炎症反应，与最近的临床前动物模型明确一致 研究结果，并强调疫苗诱导的抗原特异性Th 1应答与保护 多组分疫苗。我们将使用我们的方法描述这种免疫应答的转录组学基础。 现有的人体样本采集。我们将利用我们在人全血刺激试验方面的经验 和组织模型，以建立全血和器官型扁桃体组织模型作为我们的主要研究工具。 这种方法将使我们能够询问离体人类对整个细菌的免疫反应，培养 上清液和疫苗抗原，以发现潜在的系统性和粘膜保护相关性。这些 广泛的努力，以确定免疫途径牵连疫苗诱导的保护将通知设计 一种实用的、可扩展的、与菌株无关的全血试管内测定（类似于细胞因子释放测定）， 以及高通量功能性口腔液粘膜测定，以在即将进行的新型GAS的临床试验中进行测试 疫苗。

PROJECT SUMMARY/ABSTRACT There is renewed international momentum to develop vaccines to prevent infections caused by the highly adapted, human-restricted bacterial pathogen Group A Streptococcus (GAS). However, the lack of a known correlate of human immune protection against GAS infection and the limitations of current in vitro assays and preclinical models have impeded development of promising preclinical vaccine candidates and threatens future investment in GAS vaccine discovery and design. To overcome this roadblock, we will undertake a cross- disciplinary collaborative research program, drawing on highly relevant human samples and developing a new human ex vivo tissue model, to discover broadly applicable potential human correlates of protection that will inform design of practical immunoassays fit for deployment in clinical vaccine trials. Rather than adhering to the constraints of historical GAS immunoassays, we are explicitly targeting preferred characteristics for clinical immunogenicity assays to support vaccine development, such as practicality, accuracy and broad application to relevant syndromes and strains. GAS naturally infects humans only, therefore we will take a human-centred approach, purposefully moving away from animal models that do not adequately represent relevant complexities of the human immune response. We will apply cutting edge immunology approaches to study diverse human samples from parallel research programs, including from externally funded trials using our own GAS human challenge model to evaluate vaccine protection against pharyngitis, the primary target indication for vaccine development efforts. Initial published findings from the GAS pharyngitis human challenge model show that protection may be associated with robust early mucosal and systemic Th1 inflammatory responses, in clear alignment with recent preclinical animal model findings, and highlighting vaccine-induced antigen-specific Th1 responses as a correlate of protection for multicomponent vaccines. We will characterize the transcriptomic basis for this immune response using our existing human sample collection. We will draw on our experience with human whole blood stimulation assays and tissue models to establish whole-blood and organotypic tonsil tissue models as our primary research tools. This approach will allow us to interrogate ex vivo human immune responses to whole bacteria, culture supernatant, and vaccine antigens to discover potential systemic and mucosal correlates of protection. These broad-ranging efforts to identify immune pathways implicated in vaccine-induced protection will inform the design of a practical, scalable, and strain-agnostic whole-blood in-tube assay (analogous to cytokine release assays), and a high-throughput functional oral fluid mucosal assay to test in forthcoming clinical trials of novel GAS vaccines.