Project 3: Mechanistic studies and comparisons of vaccines in preclinical models

项目摘要-项目3：临床前疫苗的机理研究和比较 模型 该项目的两个主要目标是：(I)确定免疫学参数(即，CD4+T细胞帮助； 先天性免疫反应)，在小鼠的机制研究中调节抗体反应的广度， 和(2)评估泛肉瘤病毒和泛贝塔冠状病毒候选疫苗，旨在广泛诱导 非人灵长类(NHP)的保护性免疫反应(核心D，非人灵长类，维林格)。 在目标1中，我们将确定先天免疫系统和T滤泡细胞调节的机制 项目2中设计的广泛保护性冠状病毒疫苗诱导的抗体反应的广度 (国王)。我们将在小鼠身上进行机制研究，以确定CD4+T细胞的帮助和特异性 DC亚群在调节疫苗诱导的抗体反应广度中的作用这将产生关于关键问题的见解 确定抗体广度的免疫学参数，并展望未来，为 开发刺激广泛抗体反应的佐剂。在目标2中，我们将在卫生保健计划中进行两项研究，以 评估项目2中的泛肉瘤病毒和泛贝塔冠状病毒候选疫苗，并选择领导 进一步临床前和临床开发的候选者。这些研究将使用临床上相关的佐剂。 以确保最佳的人类翻译。我们将评估NAB的规模、广度和持久性 用AS03和3M-052/明矾配制的纳米颗粒免疫原免疫诱导的反应， 两者都表现出了优越的佐剂效果，就其促进 NHP中的高强度和持久的中和抗体反应。因此我们将直接将AS03与 3M-052/明矾增强NAB反应广度的能力。我们将与项目1合作 (Veesler)和核心B(病毒学，Baric)，C(病毒进化，Bloom)和D来评估免疫原性 以及由每个纳米颗粒疫苗候选引起的反应的保护广度，并定义表位 交叉反应反应的特异性。我们将描述抗原特异性B细胞的动力学 反应，评估浆母细胞和浆细胞频率，并对抗原特异性单个B细胞进行分类和分离 来自免疫的NHP的单抗。这些单抗将由Core A(蛋白质科学， King)，并在项目1和核心B和C中分析了它们的结合和中和宽度、表位 特异性和对逃逸突变的抵抗力。

PROJECT SUMMARY– PROJECT 3: Mechanistic studies and comparisons of vaccines in preclinical models The two primary goals of the Project are to: (i) define the immunological parameters (i.e., CD4+ T cell help; innate immune response) that regulate the breadth of the antibody response in mechanistic studies in mice, and (ii) evaluate pan-sarbecovirus and pan-betacoronavirus vaccine candidates aimed at inducing broadly protective immune responses in nonhuman primates (NHPs) (with Core D, Nonhuman Primates, Villinger). In Aim 1, we will determine the mechanisms by which the innate immune system and T follicular cells regulate the breadth of antibody responses induced by broadly protective coronavirus vaccines designed in Project 2 (King). We will perform mechanistic studies in mice to determine the impact of CD4+ T cell help and specific DC subsets in regulating the breadth of vaccine-elicited Ab responses. This will yield insights about critical immunological parameters that determine antibody breadth and, looking ahead, provide a rational basis for the development of adjuvants that stimulate broad Ab responses. In Aim 2 we will conduct two studies in NHPs to evaluate pan-sarbecovirus and pan-betacoronavirus vaccine candidates from Project 2 and select lead candidates for further preclinical and clinical development. These studies will use clinically relevant adjuvants to ensure optimal translation to humans. We will assess the magnitude, breadth and durability of nAb responses induced by immunization with nanoparticle immunogens formulated with AS03 and 3M-052/alum, both of which have demonstrated superior adjuvant effects in terms of their capacity to promote high-magnitude and durable neutralizing antibody responses in NHPs. We will thus directly compare AS03 with 3M-052/alum for their capacity to enhance the breadth of nAb responses. We will work with Project 1 (Veesler) and Cores B (Virology, Baric), C (Viral Evolution, Bloom) and D to evaluate the immunogenicity and protective breadth of the responses elicited by each nanoparticle vaccine candidate and define the epitope specificities of cross-reactive responses. We will characterize the dynamics of the antigen-specific B cell response, assess plasmablast and plasma cell frequencies, and sort antigen-specific single B cells and isolate monoclonal antibodies from immunized NHPs. These mAbs will be produced by Core A (Protein Sciences, King) and analyzed in Project 1 and Cores B and C for their binding and neutralizing breadth, epitope specificity, and resistance to escape mutations.