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，引起季节性流行， 零星的流行病新出现的人畜共患流感病毒造成的持续威胁， 免疫学上的幼稚，代表了一个主要的全球关注。目前的流感疫苗引起狭窄的，菌株- 特异性免疫，过度依赖于鸡蛋为基础的制造，有一个长期的生产过程， 同时引发对多种IAV抗原（Ag）的强细胞和体液免疫应答。的 季节性流感疫苗的有效性不一，突出了投资于早期 临床前评价可引起更广泛免疫反应的创新疫苗 针对新出现的病毒/变种。努力开发一种通用流感病毒疫苗， 针对季节性和大流行亚型的广泛和长期保护，重点是诱导免疫 针对流感病毒Ag上的高度保守表位的应答，例如主要的 表面糖蛋白血凝素（HA）、神经氨酸酶（NA）或内部核蛋白（NP）。 在R 01中，我们将开发一种创新的、优化的、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资金产生的初步数据使 鉴定第1组IAV的前导无头HA，并证明我们至少可以成功编码 两个Ag在单个表达盒中。我们现在将建立在这些数据和工程师双或三顺反子银 与NA和/或NP组合的编码我们的铅无头HA的盒。我们将扩大和扩大 免疫亚显性HA茎或长期被忽视的NA的免疫识别，通过使用融合- 基于Ag的分子佐剂方法称为“外泌体展示”，其促进Ag靶向宿主， 衍生的细胞外囊泡，包括体内的外来体。外泌体在调节细胞凋亡中起重要作用。 我们已经证明，外泌体展示可以显着增加免疫原性， 模型Ag由两种不同的腺病毒（Ad）载体平台编码。优化的“佐剂化”Ag表达 盒将被工程化到Ad载体的疫苗中，在人类中具有低血清阳性率， Ag与宿主来源的外泌体的束缚。这可以通过增加对免疫系统的识别来增强免疫反应。 被免疫系统编码的抗原最后，我们将综合评估和表型的免疫 这些通用疫苗在单次注射方案中的特征，以及在用异源性疫苗致死性攻击中的测试功效。 和异亚型IAV。在冠状病毒大流行期间，广告载体由于其 易于制造，成本低廉，热稳定性的可能性，免疫原性的损失最小 在无冷链条件下，使其成为公平全球分销的理想候选产品。因此 本R 01中描述的基于广告的通用流感疫苗将适合于大流行的储备 它可以在一次射击后提供“普遍”保护。