Globally Appropriate Genome Reduced Killed Whole Bacterial HIV Vaccines

全球适用的基因组减少灭活全细菌 HIV 疫苗

基本信息

批准号：
10672822
负责人：
Steven L. Zeichner
金额：
$ 56.53万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-13 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10672822
关键词：
2019-nCoV Adjuvant Animal Model Antibodies Antibody Binding Sites Antibody Formation Antigens Bacteria Bacterial Vaccines Binding Binding Sites Biological Assay Biomedical Engineering Biomedical Research Cells Cellular Immunity Chimeric Proteins Clinical Cold Chains Combined Vaccines Consensus Coronavirus Cryopreservation DNA DNA biosynthesis Data Development Disease Dose Engineering Enzyme-Linked Immunosorbent Assay Epidemic Escherichia coli Escherichia coli Vaccines Exhibits Family suidae Funding Future Genome Goals HIV HIV Antigens HIV vaccine HIV-1 HIV-1 vaccine Hand Human Immune response Immunize Inactivated Vaccines Industrialization International Knowledge Letters Manufacturer Medical Membrane Modeling Monitor Monoclonal Antibodies Mus Peptide Vaccines Peptides Phase Plasmids Positioning Attribute Preventive vaccine Production Proteins Recombinants Recording of previous events Research Route Safety Structure Study models Surface System T cell response Technology Transfer Testing Training Translating Vaccinated Vaccination Vaccine Antigen Vaccine Production Vaccines Viral Antigens Viral Envelope Proteins Viral Fusion Proteins Virus Virus Diseases Whole Cell Vaccine Work antigen-specific T cells cell mediated immune response clinical efficacy cost design early phase clinical trial forgiveness high reward high risk immunogenicity improved in vivo evaluation innovation manufacturing capabilities meetings monomer mouse model neutralizing antibody neutralizing monoclonal antibodies nonhuman primate novel novel strategies novel vaccines porcine epidemic diarrhea virus prophylactic response safety assessment skills success synthetic biology vaccine candidate vaccine development vaccine immunogenicity vaccine platform vaccine response

项目摘要

A broad consensus regarding HIV vaccine development calls for engineering antigens that elicit production of antibodies like the known Broadly Neutralizing (BN) Monoclonal Antibodies (MAbs). Two Env BN MAb binding sites, the Membrane Proximal External Region (MPER) and Fusion Peptide (FP), are attractive targets because they are linear peptides. We developed a new, low cost, globally appropriate vaccine platform: Killed Whole Cell (KWC) Genome-Reduced E. coli (grEc), with vaccine antigens expressed on bacterial surfaces using Gram- autotransporters. Using synthetic biology, testable vaccine candidates can be made in ~3 wks for ~$50 each. Vaccines made with the platform will cost ~$1/dose, can be produced in existing factories globally, and have forgiving cold chain requirements. We made coronavirus FP vaccines using the KWC grEc platform and showed clinical efficacy in an animal model. We propose to make KWC grEc HIV vaccines targeting MPER and FP. Our preliminary data show that we can express HIV MPER and FP Ags with this approach and elicit HIV neutralizing sera in mice using an MPER-derived Ag. We hypothesize that the KWC grEc platform, targeting MPER and FP, will yield safe, effective, low cost, globally appropriate HIV vaccines. In PHASE 1, Aim 1, we will synthesize DNAs encoding MPER and FP Ags, employing bioengineering strategies to enhance Ag exposure and antigenicity. We will clone these DNAs into our expression plasmid and transform into grEc to make KWC candidate vaccines. Using highly neutralizing and relatively non-neutralizing MAbs, we will test candidate vaccines, selecting those that show the best difference in binding neutralizing vs. less neutralizing MAbs for in vivo testing. In Aim 2, we will vaccinate mice to assess vaccine immunogenicity, comparing the different candidate vaccines, alone and in combination, to identify the best vaccines and dose/route/adjuvant using ELISAs, ELIspot assays, and PhenoSense neutralization assays against a representative panel of viruses. Decision Gate to progress to Phase 2: Production of a single or combination vaccine that elicits sera in vaccinated mice yielding an IC50 >100 in the PhenoSense neutralization assay for ≥75% of mice immunized, for ≥75% of HIV-1 Env reference strains, while sera from control immunized mice (mice immunized with bacteria not expressing viral antigen) show no neutralization above baseline control. In PHASE 2 we will, in Aim 3, conduct a non-human primate (NHP) model study to assess the safety, immunogenicity, and ability to elicit a neutralizing Ab response by the vaccines. We expect that our vaccines will induce specific HIV Ag-binding Abs, HIV antigen-specific T cell responses, and BN neutralizing Ab responses in NHP, implying that the vaccine will be safe and effective in humans. KWC vaccines have a long history, are inexpensive to make, and can be produced globally in existing facilities, we anticipate that our work can be quickly translated into safe and effective, inexpensive, globally appropriate, prophylactic HIV vaccines.

关于引起生产的工程抗原的HIV疫苗开发的广泛共识诸如已知的广泛中和（BN）单克隆抗体（mAb）之类的抗体。两个env bn mab 结合位点，膜近端外部区域（MPER）和融合肽（FP）是有吸引力的目标因为它们是线性肽。我们开发了一个新的，低成本的全球疫苗平台：被杀全细胞（KWC）基因组还原大肠杆菌（GREC），疫苗抗原在细菌表面表达使用革兰氏阴性菌。使用合成生物学，可以在〜3 wks中进行可测试的疫苗候选物每个〜$ 50。该平台制造的疫苗的价格为约1美元/剂量，可以在全球现有工厂中生产，并有宽容的冷链要求。我们使用KWC GREC平台制作了冠状病毒FP疫苗并在动物模型中显示出临床效率。我们建议将KWC GREC HIV疫苗靶向 MPER和FP。我们的初步数据表明，我们可以通过这种方法表达HIV MPER和FP AGS 使用MPER衍生的AG引起小鼠中和血清的HIV。我们假设KWC GREC平台，针对MPER和FP，将产生安全，有效，低成本，全球适合的HIV疫苗。在第1阶段，AIM 1中，我们将使用生物工程来合成编码MPER和FP AGS的DNA 增强农业暴露和抗原性的策略。我们将这些DNA克隆到我们的表达质粒中，并且转变为GREC以制造kWC候选疫苗。使用高度中和和相对非中和 mabs，我们将测试候选疫苗，选择那些在结合中和Vs中显示最佳差异的疫苗。用于体内测试的中和mAb较少。在AIM 2中，我们将接种疫苗以评估疫苗免疫原性，比较单独和组合不同候选疫苗，以识别最好的疫苗和使用ELISA，ELISPOT分析和现象识别协议测定法的剂量/路线/佐剂病毒的代表面板。进行第2阶段的决策门：产生单一或组合疫苗，引起血清中的血清接种疫苗的小鼠在现象识别神经化测定中产生IC50> 100的疫苗，以≥75％的免疫小鼠免疫，对于≥75％的HIV-1 Env参考菌株，而对照免疫抑制小鼠的血清（小鼠免疫抑制未表达病毒抗原的细菌在基线控制上没有任何谈判。在第2阶段，我们将在AIM 3中进行非人类灵长类动物（NHP）模型研究，以评估安全性，免疫原性和引起疫苗中和AB反应的能力。我们希望我们的疫苗将诱导特定的HIV AG结合ABS，HIV抗原特异性T细胞反应和BN中和AB NHP的反应意味着疫苗在人类中将是安全有效的。 KWC疫苗有很长的历史，制造便宜，可以在现有设施中在全球生产，我们预计我们的工作可以快速转化为安全有效，廉价，适合全球性的预防性HIV疫苗。