Combining innovative molecular adjuvanting approaches with novel adenoviral vector delivery to generate a universal influenza vaccine

将创新的分子佐剂方法与新型腺病毒载体递送相结合以产生通用流感疫苗

• 批准号: 10519005
• 负责人: Lynda Coughlan
• 金额: $ 52.66万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10519005
• 关键词: Address; Adenovirus Vector; Adjuvant; Adoptive Transfer; Antibodies; Antibody Response; Antigen Targeting; Antigens; B-Lymphocytes; Benchmarking; Bronchoalveolar Lavage; CD3 Antigens; CD8B1 gene; CMV promoter; Cells; Cellular Immunity; Clinical; Cold Chains; Complement; DNA cassette; Data; Data Engineering; Dose; Effectiveness; Engineering; Enzyme-Linked Immunosorbent Assay; Epidemic; Epitopes; Female; Funding; Hemagglutinin; Human; Humoral Immunities; Immune; Immune Sera; Immune response; Immune system; Immunity; Immunization; Immunoglobulin A; Immunoglobulin G; Immunologics; In Vitro; Individual; Inflammation; Influenza A virus; Intramuscular; Lead; Lung; Lymphocyte; Measures; Mediating; Membrane Glycoproteins; Modeling; Molecular; Morbidity - disease rate; Mucous Membrane; Mus; Neuraminidase; Nucleoproteins; Outcome; PTPRC gene; Peptides; Phenotype; Phylogenetic Analysis; Play; Population; Process; Production; Proteins; Publishing; Pulmonary Pathology; Readiness; Regimen; Regulation; Research; Role; Route; Safety; Seroprevalences; Serum; Spleen; Stains; System; T-Lymphocyte; Testing; Ultracentrifugation; Vaccinated; Vaccination; Vaccine Antigen; Vaccines; Validation; Variant; Viral; Viral Antigens; Virus; Work; Zoonoses; antigen-specific T cells; base; cesium chloride; comparative; cost; cost effective; cross reactivity; cytokine; delivery vehicle; draining lymph node; efficacy testing; egg; enhanced green fluorescent protein; exosome; extracellular vesicles; homologous recombination; human pathogen; immunogenic; immunogenicity; in vivo; influenza virus vaccine; influenzavirus; innovation; innovative technologies; lung histology; male; neutralizing antibody; novel; pandemic coronavirus; pandemic disease; pandemic preparedness; particle; pathogen; pre-clinical; preclinical evaluation; respiratory pathogen; response; seasonal influenza; universal influenza vaccine; universal vaccine; vaccine evaluation; vaccine platform; vaccine safety; vector; vector vaccine

SUMMARY: Influenza A viruses (IAVs) are important human pathogens, which cause seasonal epidemics and sporadic pandemics. The ongoing threat posed by emerging zoonotic influenza viruses, for which humans are immunologically naïve, represents a major global concern. Current influenza vaccines elicit narrow, strain- specific immunity, are overly reliant on egg-based manufacturing, have a prolonged production process, and fail to elicit robust cellular and humoral immune responses to multiple IAV antigens (Ags) simultaneously. The variable effectiveness of seasonal influenza vaccines, has highlighted the importance of investing in the early pre-clinical evaluation of innovative vaccines which could elicit immune responses with increased breadth against emerging viruses/variants. Efforts to develop a universal influenza virus vaccine, capable of providing broad and long-lived protection against seasonal and pandemic subtypes, are focused on inducing immune responses directed towards highly conserved epitopes on influenza virus Ags such as the stalk of the major surface glycoprotein hemagglutinin (HA), the neuraminidase (NA) or the internal nucleoprotein (NP). In this R01, we will develop an innovative, optimized, universal influenza vaccine platform to overcome issues associated with current vaccines. Preliminary data generated through R21 funding enabled the identification of lead headless HAs for group 1 IAVs, and demonstrated that we can successfully encode at least two Ags in a single expression cassette. We will now build upon these data and engineer bi- or tri-cistronic Ag cassettes encoding our lead headless HAs in combination with NA and/or NP. We will augment and broaden immune recognition of the immunosubdominant HA stalk, or long overlooked NA, by using employing a fusion- Ag based molecular adjuvanting approach called “exosome-display”, which facilitates targeting of Ags to host- derived extracellular vesicles including exosomes in vivo. Exosomes play important roles in the regulation of immunity, and we have demonstrated that exosome-display can dramatically increase the immunogenicity of a model Ag encoded by two distinct adenoviral (Ad) vector platforms. Optimized, “adjuvanted” Ag expression cassettes will be engineered into Ad vectored vaccines with low seroprevalence in humans, allowing in vivo tethering of Ag to host-derived exosomes. This could potentiate immune responses by increasing recognition of the encoded Ag by the immune system. Finally, we will comprehensively evaluate and phenotype the immune profile of these universal vaccines in single-shot regimens, and test efficacy in lethal challenge with heterologous and heterosubtypic IAVs. Ad vectors have risen to prominence during the coronavirus pandemic, due to their ease of manufacturing, cheap cost, the possibility for thermostabilization with minimal losses to immunogenicity under cold-chain free conditions, making them ideal candidates for equitable global distribution. Therefore, the Ad-based universal influenza vaccines described in this R01 would be suited to stockpiling for pandemic preparedness, and could provide “universal” protection following a single shot.

摘要：甲型流感病毒（IAV）是重要的人类病原体，可引起季节性流行和 零星流行病。新出现的人畜共患流感病毒所构成的持续威胁，人类正在面临这种威胁 免疫学上的幼稚，代表了全球的主要关注。目前的流感疫苗只能引发狭隘的、毒株- 特异性免疫，过度依赖鸡蛋制造，生产过程漫长，并且失败 同时引发针对多种 IAV 抗原 (Ag) 的强大细胞和体液免疫反应。这 季节性流感疫苗的有效性参差不齐，凸显了早期投资的重要性 创新疫苗的临床前评估，可引发更广泛的免疫反应 对抗新出现的病毒/变种。努力开发通用流感病毒疫苗，能够提供 针对季节性和大流行亚型的广泛而长期的保护，重点是诱导免疫 针对流感病毒抗原上高度保守的表位（例如主要的茎）的反应 表面糖蛋白血凝素 (HA)、神经氨酸酶 (NA) 或内部核蛋白 (NP)。 在这次R01中，我们将开发一个创新的、优化的、通用的流感疫苗平台来克服 与当前疫苗相关的问题。通过 R21 资助生成的初步数据使 识别第 1 组 IAV 的主要无头 HA，并证明我们至少可以成功编码 单个表达盒中的两个 Ag。我们现在将基于这些数据并设计双顺反子或三顺反子 Ag 编码我们的领先无头 HA 与 NA 和/或 NP 组合的盒。我们将扩大和扩大 通过使用融合-免疫显性HA茎或长期被忽视的NA的免疫识别 基于 Ag 的分子佐剂方法称为“外泌体展示”，该方法有助于将 Ag 靶向宿主 衍生的细胞外囊泡，包括体内的外泌体。外泌体在调节中发挥重要作用 免疫，我们已经证明外泌体展示可以显着提高免疫原性 由两个不同的腺病毒 (Ad) 载体平台编码的模型 Ag。优化的“辅助”Ag 表达 盒将被设计成在人类中血清阳性率较低的 Ad 载体疫苗，从而允许在体内 Ag 与宿主衍生的外泌体的束缚。这可以通过增加对以下物质的认识来增强免疫反应 免疫系统编码的Ag​​。最后，我们将对免疫进行综合评估和表型分析。 这些通用疫苗在单次注射方案中的概况，并测试异源疫苗致死攻击的功效 和异亚型 IAV。广告载体在冠状病毒大流行期间变得越来越重要，因为它们 易于制造、成本低廉、热稳定的可能性且免疫原性损失最小 在无冷链条件下，使它们成为公平全球分配的理想选择。因此， 本 R01 中描述的基于广告的通用流感疫苗适合为大流行而储存 准备，并可以在一次射击后提供“普遍”保护。