Understanding Human B Cell Responses to Influenza to Improve the Influenza

我们有令人信服的新数据表明，在不同的情况下提供流感疫苗可以诱导长期B细胞记忆流感的显着变化。这一重要的概念证明表明，可以根据需要重新训练人类B细胞反应，以提高疫苗功效。在这个项目中，我们将探索这些观察的机制。收集到的原理将对改进流感疫苗的开发非常重要。首先，在目的1中，我们将探索B细胞记忆和免疫血清在重新暴露于变异流感毒株后将抗体应答重定向至特定表位中的作用。其次，在目标2中，我们将确定在首次暴露于2009年流感大流行株时，是否像对血凝素的反应一样，广泛交叉反应的B细胞的优势被激活以对抗神经氨酸酶。我们还将使用新的方法来确定先前观察到的在人群中的偏向性流感表位靶向的稳定性。最后，我们已经确定了我们认为是"免疫记忆"的B细胞分化，我们假设这种分化以高数量持续数月而不是永久性的，并且由于在流行病期间极有可能发生的再暴露而提供了立即的保护。我们认为，这些记忆细胞，当诱导相关或不相关的菌株接种疫苗，可以在很大程度上影响在不久的将来发生的变异流感病毒株的反应，在原始抗原sin（OAS）的方式。事实上，这些细胞可能解释了我们在队列中的一些但不是所有成员中观察到的戏剧性变化，导致反应具有更大的广度或更多的菌株特异性反应。我们将探索这些细胞的表型和功能。此外，通过确定来自该群体的B细胞的特异性，我们将直接确定这些记忆细胞在再次暴露于不同流感毒株时的影响。了解B细胞记忆的类型和阶段将有助于更好地靶向流感疫苗反应。最后，在这些研究的背景下，我们将开发新的技术，如我们的"表位阴影"的方法，使用我们的新的mAb微阵列平台，以允许快速和全球筛选单克隆和多克隆（血清）表位靶向。我们还将产生一大批抗流感的人单克隆抗体。这些抗体可以识别对开发广泛保护性疫苗重要的表位，或者mAb本身可能具有治疗潜力。

We have compelling new data demonstrating that providing the influenza vaccine in different contexts can induce dramatic shifts in long-term B cell memory to influenza. This important proof of concept suggests that human B cell responses can be re-trained as needed to improve vaccine efficacy. In this project we will explore the mechanism of these observations. The principles gleaned will be important in the development of an improved influenza vaccine. First, in Aim 1, we will explore the role of B cell memory and immune sera in redirecting antibody responses to particular epitopes upon re-exposure to variant influenza strains. Secondly, in Aim 2 we will determine if, like the response to hemagglutinin, a predominance of broadly cross-reactive B cells were activated against neuraminidase upon first exposure to the 2009 pandemic strain of influenza. We will also use novel approaches to determine how stable previously observed biased influenza epitope targeting in human populations is. Finally, we have identified what we believe is an "epidemic-memory" B cell differentiation that we hypothesize persists at high numbers for months rather than permanently, and provides immediate protection due to the highly likely re-exposure that will occur during an epidemic. We believe that these memory cells, when induced by vaccination with relevant or irrelevant strains, can substantially impact responses to variant influenza strains occurring in the near future in an original-antigenic sin (OAS) fashion. In fact, these cells may account for the dramatic shifts that we observed for some but not all members of our cohort, leading to responses with either greater breadth or more strain-specific responses. We will explore the phenotype and function of these cells. Further, by determining the specificity of the B cells from this population we will directly determine the impact of these memory cells upon re-exposure to a divergent influenza strain. Understanding the types and stages of B cell memory will allow better targeting of the influenza vaccine response. Finally, in the context of these studies we will develop new technologies such as our "epitope-shadowing" approach using our novel mAb microarray platform to allow rapid and global screening of both monoclonal and polyclonal (serum) epitope targeting. We will also generate a large panel of human mAbs against influenza. These antibodies may identify epitopes important for developing a broadly protective vaccine or the mAbs themselves may have therapeutic potential.