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; 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：流感病毒（IAV）是重要的人类病原体，引起季节性流行病和 零星的大流行。新兴的人畜共患影响病毒构成的持续威胁，人类是 在免疫学上，代表了全球的主要关注点。当前影响疫苗会引起狭窄的应变 - 特定的免疫力，对基于鸡蛋的制造过度敏感，生产过程延长，失败 简单地引起对多种IAV抗原（AGS）的强大细胞和体液免疫反应。这 季节性影响疫苗的可变有效性，强调了在早期投资的重要性 对创新疫苗的临床前评估，这些疫苗可能会引起宽度增加的免疫调查 反对新兴病毒/变体。开发普遍影响病毒疫苗的努力，能够提供 针对季节性和大流行亚型的广泛而长期的保护，专注于诱导的免疫 针对高度保守的表位对影响力的病毒Ag（例如主要的茎）的反应 表面糖蛋白血凝素（HA），神经瘤酶（NA）或内核蛋白（NP）。 在此R01中，我们将开发一个创新，优化，普遍的影响器疫苗平台来克服 与当前疫苗相关的问题。通过R21资金生成的初步数据启用了 针对第1组IAV的Lead Leadless具有识别，并证明我们至少可以成功编码 单个表达盒中的两个AGS。现在，我们将基于这些数据和工程师双或三个文章AG 编码我们的铅无头的录音带与NA和/或NP结合使用。我们将扩大和扩大 通过使用融合 - 基于Ag的分子调节方法称为“外泌体 - 播放”，这有助于将AG靶向宿主 - 衍生的细胞外蔬菜，包括体内外泌体。外泌体在调节中起重要作用 免疫力，我们已经证明了外泌体播放可以显着提高A的免疫原性 模型AG由两个不同的腺病毒（AD）矢量平台编码。优化的“佐剂” Ag表达 盒式磁带将被设计成具有人类血清阳性较低的AD矢量疫苗，使体内允许体内 将AG绑定到宿主衍生的外泌体。这可能通过增加对 免疫系统编码的AG。最后，我们将全面评估和表型免疫。 这些通用疫苗在单发方案中的概况，并在致命挑战中测试效率 和异源性IAV。由于它们 易于制造，便宜的成本，恒温化的可能性，免疫原性损失最小 在无冷链条件下，使其成为公平全球分布的理想候选人。因此， 基于AD的R01中描述的普遍影响将适合于大流行库存 准备，并可以在一次镜头后提供“通用”保护。