Immunity in the Nose to inFLUENza: Correlates of Efficacy (INFLUENCE)

上呼吸道是呼吸道病毒（如流感）的初始感染部位。 在这个粘膜部位的T细胞和抗体形成局部免疫记忆， 提供杀菌免疫力。尽管这种已知的重要性，在预防感染，粘膜 相对于其外周血对应物，免疫性研究不足。这导致了一个弱 了解粘膜免疫对预防病毒感染的贡献，以及 这些因素可以在下一代流感疫苗中得到增强。这项建议旨在 解决了阻碍我们理解粘膜的两个主要技术限制 免疫力首先，研究缺乏经过充分验证和一致应用的工具 呼吸道粘膜取样鼻吸附是一种微创，耐受性好， 采样工具，产生高度可重复的数据，丰富的蛋白质介质中， 人的鼻子在这里，鼻吸附将用于收集健康人的呼吸道分泌物， 对照志愿者，并与人类感染产生的生物库样本一起分析 流感的挑战研究。该第一个目的试图证明鼻吸附作为一种生物活性剂的效用。 人类鼻腔采样工具，可以为未来的免疫学带来更高水平的一致性， 疫苗效力研究。其次，研究无法确定 粘膜抗体的功能活性。这种无能是由于研究的重点是 全身免疫，其中抗体滴度高于粘膜分泌物。现有 血凝抑制、中和和抗体介导的 因此，细胞毒性测定要么不够灵敏，要么没有优化用于 粘膜样本。为了解决这一技术差距，本提案旨在开发使用 假病毒中和和Fcγ R-二聚体结合试验，以研究粘膜 抗流感抗体。这些技术非常适合粘膜样品，因为它们 比传统方法具有更高的灵敏度，可以很容易地进行修改，以研究新的 抗原随着病毒的进化和疫苗的出现。利用人类感染的样本 用野生型（H1N1）流感病毒和疫苗病毒（LAIV）样本进行的攻毒研究使 在严格控制的环境中测试这些技术，并对同性恋和异性恋进行分析， 由感染/疫苗接种引起的亚型免疫应答。最后， 将对开发的技术进行测试，以便能够广泛传播这些技术， 使得能够在未来的流感疫苗研究中理解粘膜免疫。

The upper respiratory tract is the site of initial infection with respiratory viruses, such as influenza. T cells and antibodies at this mucosal site form local immune memory and are positioned to provide sterilizing immunity. Despite this known importance in preventing infections, mucosal immunity is understudied relative to its peripheral blood counterpart. This has resulted in a weak understanding of the contribution of mucosal immunity to preventing viral infection, and those factors that could be enhanced in next-generation influenza vaccines. This proposal aims to address two major technological limitations that have hindered our understanding of mucosal immunity. Firstly, studies have suffered from a lack of well validated and consistently applied tools for sampling the respiratory mucosa. Nasosorption is a minimally invasive, well tolerated, sampling tool that yields highly reproducible data on the abundance of protein mediators in the human nose. Here, nasosorption will be employed to collect respiratory secretions from healthy control volunteers and analyzed alongside biobanked samples arising from human infection challenge studies of influenza. This first aim seeks to demonstrate the utility of nasosorption as a human nasal sampling tool that can bring a greater level of consistency to future immunology and vaccine efficacy studies. Secondly, studies have suffered from an inability to determine the functional activity of mucosal antibodies. This inability has arisen from the focus of research on systemic immunity, where antibody titres are higher than in mucosal secretions. Existing technologies such as haemagglutination inhibition, neutralization, and antibody-mediated cytotoxicity assays are therefore either insufficiently sensitive or not optimized for use with mucosal samples. To address this technology gap, this proposal aims to develop the use of pseudovirus neutralization and FcγR-dimer binding assays to study the activity of mucosal antibodies against influenza. These technologies are ideally suited to mucosal samples as they have greater sensitivity than classical approaches and can be readily modified to study new antigens as viruses evolve and vaccines emerge. Utilizing samples from human infection challenge studies with wild-type (H1N1) influenza and vaccine-virus (LAIV) samples enables the testing of these technologies in a tightly controlled setting and analysis of the homo- and hetero- subtypic immune responses resulting from infection/vaccination. Finally, the robustness of the developed technologies will be tested to enable widespread dissemination of the techniques, enabling the understanding of mucosal immunity in future studies of influenza vaccines.