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 零星的流行病。新出现的人畜共患流感病毒构成的持续威胁，对此人类是 免疫幼稚，代表了一个主要的全球关切。目前的流感疫苗引发了狭窄的、株系- 特定免疫力，过度依赖以鸡蛋为基础的制造，生产过程延长，并失败 同时诱导对多种IAV抗原的细胞和体液免疫反应。这个 季节性流感疫苗的可变效力，突显了在早期投资的重要性 可引起更广泛免疫反应的创新疫苗的临床前评估 抵御新出现的病毒/变种。努力开发一种通用流感病毒疫苗，能够提供 针对季节性和大流行亚型的广泛和持久的保护，重点是诱导免疫 针对流感病毒AGS上高度保守的表位的反应，如主要 表面糖蛋白血凝素(HA)、神经氨酸酶(NA)或内部核蛋白(NP)。 在本次R01中，我们将开发一个创新、优化、通用的流感疫苗平台，以克服 与当前疫苗相关的问题。通过R21基金产生的初步数据使 鉴定了第1组IAV的无头铅HAS，并证明我们至少可以成功编码 在一个表达盒中有两个AGs。我们现在将在这些数据的基础上设计出双顺反子或三顺反子银 编码我们的无头铅的磁带与NA和/或NP结合在一起。我们将扩大和扩大 免疫抑制的HA茎，或长期被忽视的NA的免疫识别，通过使用融合- 以银为基础的分子佐剂方法被称为外切体展示，它促进了AGS对宿主的靶向。 体内衍生的胞外囊泡，包括外切小体。外切体在细胞周期调控中发挥重要作用。 免疫，我们已经证明，外显体展示可以显著提高免疫原性 模型Ag由两个不同的腺病毒(Ad)载体平台编码。优化的、“辅助性”的抗原表达 盒式疫苗将被改造成在人体内具有低血清阳性反应率的Ad载体疫苗，允许在体内 Ag与宿主来源的外切体的连接。这可以增强免疫反应，通过增加对 免疫系统编码的抗原。最后，我们将对免疫进行全面的评估和表型 这些通用疫苗在单针方案中的概况，以及在与异源疫苗的致死攻击中的测试效果 和异亚型IAV。在冠状病毒大流行期间，腺病毒载体已经上升到突出地位，因为它们 易于制造，成本低廉，热稳定的可能性，免疫原性损失最小 在无冷链的条件下，使它们成为公平全球分配的理想候选者。因此， 本R01中描述的基于AD的通用流感疫苗将适合储备以备大流行 防备，并可在一次射击后提供“普遍”保护。