Vaccinating at Mucosal Surfaces with Nanoparticle Conjugated Antigen and Adjuvant

用纳米颗粒缀合抗原和佐剂在粘膜表面进行疫苗接种

• 批准号: 8974276
• 负责人: SEBASTIAN JOYCE
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8974276
• 关键词: Adjuvant; Antibodies; Antigen-Presenting Cells; Antigens; Attenuated; B-Lymphocytes; Biological Warfare; CD8B1 gene; Cells; Communicable Diseases; Coupled; Data; Dendritic Cells; Development; Ectromelia; Engineering; Epitopes; Formulation; Galactosylceramides; Generations; Glycolipids; Granzyme; HLA-A gene; Health; Human; Immune; Immune system; Immunity; Immunization; Indigenous; Infectious Agent; Infectious Ectromelia; Interferons; Interleukin-2; Kinetics; Learning; Life; Lipids; Lung; Mediating; Memory; Methods; Microbe; Modeling; Modified Vaccinia Virus Ankara; Mouse Pox Virus; Mus; Natural Killer Cells; Peritoneal; Personal Satisfaction; Plague; Poxviridae; Primary Infection; Process; Protein Subunits; Proteins; Proteomics; Role; Route; Smallpox; Smallpox Vaccine; Smallpox Viruses; Subunit Vaccines; Surface; T cell response; T-Lymphocyte; T-Lymphocyte Epitopes; T-Lymphocyte Subsets; TNF gene; Testing; Transgenic Mice; Unemployment; Vaccinated; Vaccination; Vaccine Design; Vaccines; Vaccinia virus; Vaccinium; Validation; Variant; Veterans; Viral Proteins; Virulent; Virus; Work; antigen processing; antimicrobial; arm; base; clinical practice; design; exhaust; expectation; humanized mouse; immunogenicity; in vivo; infectious disease model; innovation; insight; microbial; mouse model; mucosal site; mucosal vaccine; nanoparticle; next generation; novel; pathogen; protective efficacy; respiratory; response; vaccination schedule; vaccination strategy; vaccine development; vaccine efficacy

DESCRIPTION (provided by applicant): The objective of this revised VA Merit Proposal is to design CD8+ T cell-targeted pathogen- free (subunit) vaccines against infectious diseases and to devise an efficacious method for their delivery to mucosal sites, which mark the major port of pathogen entry. Most currently used vaccines work by antibody-mediated neutralization and/or opsonization. Despite the proven critical role for CD8+ T cells in protection against infectious diseases caused by intracellular microbes, effective vaccines that target this T cell subset currently do not exist. This is because naturally processed and presented CD8+ T cell epitopes and those that confer protective immunity remain unknown. To fill this gap, using the most successful smallpox vaccine as a model, we first discovered and characterized numerous HLA- A*02;01 and -B*07;01 class I restricted, naturally processed CD8+ T cell-epitopes derived from vaccinia virus (VacV) that are also conserved in variola virus-the agent of smallpox. We then found that prime-boost vaccination of mice with engineered VacV proteins containing such epitopes in combination with an NKT cell-targeted adjuvant �- galactosylceramide (�GC)-which rapidly activates NKT cells and potently trans-activates dendritic cells as well as natural killer, T and B cells-elicited robut, functional CD8+ T cell responses. These CD8+ T cell responses protected mice from lethal respiratory poxvirus challenge. Guided by these findings, we will test the central hypothesis that, "Nanoparticle- based mucosal delivery of microbial protein subunit vaccines formulated with an optimal adjuvant confers protective immunity against lethal respiratory challenge by targeting naturally processed CD8+ T cell epitopes". Our approaches to test the central hypothesis are: (a) to prepare nanoparticles (np) coupled with �GC (GC-np) or the IFNy-inducing GC variant, flurobenzyl-acyl-�GC (FBzGC-np), and to characterize the mechanism of their action; (b) to determine the quality and robustness of pulmonary CD8+ T cell responses to immunization with nanoparticles that will co-deliver coupled recombinant VacV subunit(s)- np and GC-np or FBzGC-np; and (c) to elucidate the protective potential of such a vaccine strategy against lethal respiratory challenge with heterotypic (VacV-WR strain) and homotypic (Ectromelia) poxvirus models. The approaches proposed herein are feasible because our new data revealed that rVV-np+GC-np elicited better protective response in humanized mice upon intranasal vaccination than intra-peritoneal immunization. Hence, we have proposed innovative approaches to develop microbe-free, mucosal vaccines containing naturally processed antigens and NKT cell-targeted adjuvant, which when delivered on nanoparticle carriers, mediate protection through CD8+ T cells. Such mucosal vaccine formulation targets previously unemployed, yet greatly potent arms of the immune system critical for protection against intracellular infectious disease agents-viz., the innate-like NKT cells and the adaptive CD8+ T cells. Because the vaccine will be tested in both heterotypic and homotypic pathogen-host models and a humanized mouse model, we expect to gain novel insights relevant to the design of next generation pathogen-free vaccines. Such vaccines will have the potential to shift clinical practice paradigms, especially against infectious diseases that directly impact the health and well-being of this World's peoples, and, hence, will benefit Veterans as well.

描述（由申请人提供）： 修订后的 VA 优秀提案的目的是设计针对传染病的 CD8+ T 细胞靶向无病原体（亚单位）疫苗，并设计一种有效的方法将其递送至粘膜部位，粘膜部位是病原体进入的主要端口。目前使用的大多数疫苗通过抗体介导的中和作用和/或调理作用发挥作用。尽管已证明 CD8+ T 细胞在预防细胞内微生物引起的传染病方面发挥着关键作用，但目前尚不存在针对该 T 细胞亚群的有效疫苗。这是因为 自然加工和呈递的 CD8+ T 细胞表位以及那些赋予保护性免疫的表位仍然未知。为了填补这一空白，我们以最成功的天花疫苗为模型，首先发现并表征了许多 HLA-A*02;01 和 -B*07;01 I 类限制性、自然加工的 CD8+ T 细胞表位，这些表位源自痘苗病毒 (VacV)，这些表位在天花病毒天花病毒中也保守。然后我们发现，用含有此类表位的工程化 VacV 蛋白与 NKT 细胞靶向佐剂半乳糖神经酰胺 (GC) 相结合，对小鼠进行初免加强疫苗接种，该佐剂可快速激活 NKT 细胞并有效反式激活树突状细胞以及自然杀伤细胞、T 细胞和 B 细胞，从而引发强大的功能性 CD8+ T 细胞反应。这些 CD8+ T 细胞反应保护小鼠免受致命的呼吸道痘病毒的攻击。在这些发现的指导下，我们将检验中心假设： “基于纳米粒子的粘膜递送微生物蛋白亚单位疫苗，采用最佳佐剂配制，通过靶向自然加工的 CD8+ T 细胞表位，赋予针对致命呼吸道挑战的保护性免疫力”。我们测试中心假设的方法是：（a）制备与αGC（GC-np）或IFNγ诱导GC变体氟苄基-酰基-αGC（FBzGC-np）偶联的纳米颗粒（np），并表征其作用机制； (b) 确定肺 CD8+ T 细胞对纳米颗粒免疫反应的质量和稳健性，纳米颗粒将共同递送偶联的重组 VacV 亚基-np 和 GC-np 或 FBzGC-np； (c) 阐明这种疫苗策略对异型（VacV-WR 株）和同型（Ectromelia）痘病毒模型的致命性呼吸道攻击的保护潜力。本文提出的方法是可行的，因为我们的新数据显示，与腹膜内免疫相比，rVV-np+GC-np 在人源化小鼠中鼻内疫苗接种后引起更好的保护反应。因此，我们提出了创新方法来开发含有天然加工抗原和 NKT 细胞靶向佐剂的无微生物粘膜疫苗，当将其递送到纳米颗粒载体上时，通过 CD8+ T 细胞介导保护。这种粘膜疫苗制剂的目标是以前未使用但非常有效的免疫系统臂，对于防御细胞内传染病病原体（即先天样 NKT 细胞和适应性 CD8+ T 细胞）至关重要。由于该疫苗将在异型和同型病原体宿主模型以及人源化小鼠模型中进行测试，因此我们期望获得与下一代无病原体疫苗设计相关的新见解。此类疫苗将有可能改变临床实践范式，特别是针对直接影响世界人民健康和福祉的传染病，因此也将使退伍军人受益。