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

上呼吸道是最初感染呼吸道病毒的地方，如流感。 这个粘膜部位的T细胞和抗体形成局部免疫记忆，并定位为 提供消毒免疫力。尽管已知在预防感染方面的重要性，但粘膜 相对于外周血液，免疫力的研究还不够深入。这导致了一个疲软的 了解粘膜免疫在预防病毒感染中的作用，以及 下一代流感疫苗中可以增强的因素。这项建议旨在 解决阻碍我们理解粘膜的两个主要技术限制 豁免权。首先，研究缺乏经过充分验证和一致应用的工具。 对呼吸道粘膜进行采样。鼻吸是一种微创、耐受性好的 一种采样工具，可产生关于蛋白质介体丰度的高度重复性数据 人类的鼻子。在这里，鼻吸将被用来收集健康人的呼吸道分泌物 对照志愿者并与来自人类感染的生物库样本一起进行分析 挑战有关流感的研究。这第一个目的是试图证明鼻吸附剂作为一种 人类鼻腔采样工具，可以为未来的免疫学和 疫苗效力研究。其次，研究无法确定 粘膜抗体的功能活性。这种无能为力的原因是人们对 全身性免疫，抗体滴度高于粘膜分泌物。现有 血凝抑制、中和和抗体介导等技术 因此，细胞毒性检测要么不够灵敏，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 用假病毒中和试验和Fc-γR-二聚体结合试验研究粘膜活性 抗流感抗体。这些技术非常适合于粘膜样本，因为它们 具有比传统方法更高的敏感性，并且可以很容易地进行修改以研究新的 随着病毒的进化和疫苗的出现，抗原也随之产生。利用人类感染样本 对野生型(H1N1)流感和疫苗病毒(LAIV)样本的挑战研究使 在严格控制的环境中测试这些技术，并分析同质和异质- 感染/接种引起的亚型免疫反应。最后，它的健壮性 将对开发的技术进行测试，以使这些技术能够广泛传播， 使我们能够在未来的流感疫苗研究中了解粘膜免疫。

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.