Immunogenicity to B and T cell vaccines in non-human primates (NHPs)

摘要--项目3 项目3的目标是评估非人灵长类(NHP)的免疫策略，目的是 在肝脏中诱导丙型肝炎病毒特异性的bNAbs和TRMS。特别是，我们将评估其规模和 用佐剂免疫丙型肝炎病毒E1/E2抗原诱导的bNab反应的持久性； 表达丙型肝炎病毒的异源病毒载体序贯免疫诱导的T细胞应答 NS3-NS5B嵌合体抗原。我们最近的工作证明了TLR7/8配体3M-052的佐剂能力 有效地刺激强健和持久的NAB反应以及显著长寿的浆细胞 (LLPC)在骨髓中，类似于观察到的活病毒疫苗的反应。因此，在目标1中，我们将 评价3M-052/明矾对丙型肝炎病毒E1/E2特异性高强度和持久性的刺激能力 BNab响应。作为对照，我们将使用一种新型佐剂，由脂质体、TLR4激动剂组成 (单磷酰脂A，MPL)和称为QS-21的皂苷(Lipo/QS-21/MPL)，这是最近显示的 在小鼠体内诱导高水平的丙型肝炎病毒糖蛋白特异性T细胞反应。 目的1：测定3M-052/明矾和Lipo/QS-21/MPL对佐剂抗原的杀伤能力。 对丙型肝炎病毒E1/E2的特异性免疫应答。在这个目标中，我们将检验这样一个假设，即使用 加佐剂的丙型肝炎病毒E1/E2可在NHP中产生强健和持续的NAB应答。 在T细胞的情况下，我们最近的结果表明，疫苗接种诱导了CD8+TRMS和 NAB反应可以协同作用，针对黏膜感染的病毒感染提供增强的保护。 利用单细胞转录图谱，我们证明了粘膜组织中TRMS的重新激活 刺激粘膜髓系细胞中的抗病毒限制因子，从而创造一个限制性的局部环境 用于病毒入侵。我们假设，诱导抗原特异性肝脏TRMS的丙型肝炎疫苗和 Nabs将通过类似的涉及本地天生的机制来提供对丙型肝炎病毒的更好的保护。 肝中髓系细胞和肝细胞的抗病毒反应。 目的2：表征序贯免疫诱导的先天和获得性免疫反应 表达丙型肝炎病毒NS3-NS5B嵌合体抗原的异源病毒载体免疫 用E1/E2和NS3-5抗原加佐剂免疫。我们将按顺序对NHP进行免疫 Ad48和MVA表达嵌合体NS3-5(诱导NS3-5特异性CD8+T细胞)，免疫后 使用可溶性丙型肝炎病毒E1/E2抗原(用于诱导NAB)和可溶性NS3-5抗原(用于促进CD8+T细胞 由病毒载体启动的应答)，给予佐剂。我们将分析先天的和适应性的 在三个分目标中作出回应。

ABSTRACT - PROJECT 3 The goal of Project 3 is to assess in nonhuman primates (NHPs), immunization strategies aimed at inducing HCV-specific bnAbs and TRMs in the liver. In particular, we will assess the magnitude and durability of bnAb responses induced by immunization with HCV E1/E2 antigens administered with adjuvants; and T cell responses induced by sequential immunization with heterologous viral vectors expressing HCV NS3-NS5b mosaic antigens. Our recent work has demonstrated the adjuvant capacity of TLR7/8 ligand 3M-052 to potently stimulate robust and durable nAb responses as well as remarkably long-lived plasma cells (LLPCs) in bone marrow, similar to responses observed with live viral vaccines. Therefore, in Aim 1 we will evaluate the capacity of 3M-052/alum to stimulate a high magnitude and durability of HCV E1/E2 specific bnAb responses. As a comparator, we will use a novel adjuvant, composed of a liposome, a TLR4 agonist (monophosphoryl lipid A, MPL) and the saponin called QS-21, (Lipo/ QS-21/ MPL), which was recently shown to induce a superior HCV glycoprotein-specific T cell response in mice. Aim 1: To determine the capacity of 3M-052/alum and Lipo/QS-21/MPL to adjuvant antigen- specific immune responses to HCV E1/E2. In this aim, we will test the hypothesis that immunization with HCV E1/E2 with adjuvant results in robust and sustained nAb responses in NHPs. In the case of T cells, our recent results demonstrate that vaccination induced CD8+ TRMs and nAb responses can synergize to provide enhanced protection against viral acquisition to mucosal infectioN. Using single cell transcriptional profiling we demonstrated that reactivation of TRMs in mucosal tissues stimulates anti-viral restriction factors in mucosal myeloid cells, thereby creating a restrictive local environment for viral entry. We hypothesize that an HCV vaccine that induces antigen-specific liver TRMs and nAbs will confer superior protection against HCV, through a similar mechanism involving local innate antiviral responses in myeloid cells and hepatocytes in the liver. Aim 2: To characterize the innate and adaptive immune responses induced by sequential immunization with heterologous viral vectors expressing HCV NS3-NS5b mosaic antigens, followed by immunization with E1/E2 and NS3-5 antigens plus adjuvant. We will immunize NHPs sequentially with Ad48 and MVA expressing Mosaic NS3-5, (to induce NS3-5 specific CD8+ T cells), followed by immunization with soluble HCV E1/E2 antigens (to induce nAbs), and soluble NS3-5 antigens (to boost the CD8+ T cell response primed by viral vectors), administered with an adjuvant. We will analyze the innate and adaptive response in three sub-aims.