Dendritic Cell Targeting Enhances Flavivirus Vaccine Efficacy

树突状细胞靶向增强黄病毒疫苗功效

• 批准号: 7500651
• 负责人: Gregg N. Milligan
• 金额: $ 22.1万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7500651
• 关键词: Address; Animals; Antibodies; Antibody Formation; Antigens; Antiviral Agents; Appearance; B-Lymphocytes; Biological Assay; Brefeldin A; Categories; Cell Line; Cells; Confocal Microscopy; Cytolysis; Data; Dendritic Cells; Dengue; Detection; Development; Disease; Enzyme-Linked Immunosorbent Assay; Equilibrium; Exposure to; Family; Flavivirus; Flavivirus Infections; Frozen Sections; Genes; Genome; Goals; Hand; Harvest; Human; Immune; Immune response; Immune system; Immunity; Immunohistochemistry; In Situ Hybridization; In Vitro; Infection; Interferon-alpha; Interferons; Kinetics; Knockout Mice; Knowledge; Langerhans cell; Lead; Life; Link; Measures; Messenger RNA; Mus; Names; Nature; Numbers; Pathway interactions; Production; Property; Protein Biosynthesis; RNA; Recruitment Activity; Research; Research Project Grants; Role; Signal Pathway; Site; Skin; Surface; System; T-Cell Activation; T-Lymphocyte; Technology; Testing; Tissues; Vaccinated; Vaccination; Vaccines; Viral; Viral Antigens; Virus; Virus-like particle; West Nile virus; base; cell mediated lymphocytolysis test; cytokine; improved; in vivo; insight; lymph nodes; macrophage; man; migration; monocyte; novel vaccines; particle; receptor; receptor binding; research study; response; tetramethylrhodamine isothiocyanate; uptake; vaccine development; vaccine efficacy

Description (provided by applicant): Viruses in the flavivirus family populate all three of NIAID's Category A, B, and C lists of viruses. Vaccines are needed for multiple flavivirus diseases, most notably dengue, and detailed knowledge of how flaviviruses interact with the innate immune system is critical for vaccine development. To address this need, we have developed a system to produce flavivirus virus-like particles (VLPs). VLPs are packaged, gene-deleted flaviviruses capable of initiating a replication cycle in cells in culture. VLPs are identical to normal flaviviruses in many aspects of infection, namely: receptor binding, uptake, RNA release, and RNA and nonstructural protein synthesis. However, VLP-infected cells do not produce functional virus capable of spreading to other cells. In addition to VLPs, we have produced a second type of flavivirus particle named RepliVAX. RepliVAX displays the non-spreading properties of VLPs, but in addition to encoding the viral products made by VLPs, RepliVAX also produces a sub-viral particle (SVP) with demonstrated capacity to induce antiviral immunity in man. RepliVAX is remarkably potent in mice and is being developed as a vaccine. Mice inoculated in the footpad with VLPs produce high levels of IFNa, in contrast to mice inoculated with UV-inactivated VLPs, which do not produce any IFNa. Draining lymph nodes (LN) harvested from VLP- inoculated mice contain WNV antigen and genome, and high levels of IFNa mRNA. These in vivo results suggest that VLPs are targeted to the draining LN where they induce IFNa synthesis. In vitro, VLP (and WNV) infection of cell lines derived from non-immune system cells induces production of a different type I IFN subtype (IFN¿), and induction is dependent on viral replication. IFN induction by WNV infection of human monocyte-derived dendritic cells (mDCs) is also dependent on viral replication, but the IFN produced by mDCs is of the IFNa subtype. On the other hand, human plasmacytoid DCs (pDCs), induce high levels of IFNa in response to exposure to live or inactivated WNV. Intriguingly, WNV does not productively infect pDCs. We hypothesize that WNV infects DCs (likely Langerhans cells) in the periphery, leading to their migration to the LN where they activate pDCs and recruit T and B cells. Further, we hypothesize that effective engagement of DC-centered innate signaling pathways contributes to the potency of our RepliVAX vaccine. To address these hypotheses, we will identify the precise nature of RepliVAX-infected cells in LN, determine if these cells (or neighboring cells) are responsible for the IFN production, and examine the effect of activation of these cells on the immune response to RepliVAX administration. These studies will provide important insights into links between innate and adaptive immunity that lead to the production of protective immunity to flavivirus infection and vaccination.

描述（由适用提供）：Flavivirus家族中的病毒填充了Niaid的A类，B和C的所有三个病毒列表。多种黄病毒疾病（最著名的是登革热）需要疫苗，并且有关黄酮病毒如何与先天免疫系统相互作用的详细知识对于疫苗开发至关重要。为了满足这一需求，我们开发了一种系统来生产类似黄病毒病毒的颗粒（VLP）。 VLP是包装的，基因删除的黄病毒，能够在培养中引发细胞的复制周期。在感染的许多方面，VLP与正常的黄病毒相同，即受体结合，摄取，RNA释放以及RNA和Nostructural蛋白合成。但是，VLP感染的细胞不会产生能够扩散到其他细胞的功能病毒。除了VLP之外，我们还产生了第二种名为“ deplivax”的黄病毒粒子。 REPLIVAX显示了VLP的非膨胀特性，但是除了编码VLP制造的病毒产品外，还原还会产生一个次病粒子（SVP），具有诱导人类抗病性免疫学的能力。在小鼠中，复制具有极大的潜力，并且正在作为一种疫苗开发。与紫外线灭活的VLP接种的小鼠相比，用VLP接种的小鼠会产生高水平的IFNA，而这不会产生任何IFNA。从含有WNV抗原和基因组的VLP接种小鼠以及高水平的IFNA mRNA收获的排水淋巴结（LN）。这些体内结果表明，VLP针对的是影响IFNA合成的排水LN。在体外，源自非免疫系统细胞的细胞系的VLP（和WNV）感染会影响不同类型IFN亚型的产生（IFN¿），并且诱导取决于病毒复制。 WNV感染人类单核细胞衍生的树突状细胞（MDC）的IFN诱导也取决于病毒复制，但是MDCS产生的IFN是IFNA亚型的。另一方面，人浆细胞类动物DC（PDC）会诱导高水平的IFNA，以响应暴露于活或灭活的WNV。有趣的是，WNV不会有效地感染PDC。我们假设WNV感染DC（可能是Langerhans细胞）在周围，导致它们迁移到LN，并在其中激活PDC并募集T和B细胞。此外，我们假设以DC为中心的先天信号通路的有效参与有助于我们的复制疫苗的效力。为了解决这些假设，我们将确定LN中重建细胞的确切性质，确定这些细胞（或相邻细胞）是否负责IFN的产生，并检查这些细胞激活对重复给药的免疫响应的激活影响。这些研究将提供有关先天性和适应性免疫学之间联系的重要见解，从而导致免疫学生产给黄病毒感染和疫苗接种。