Vaccine targeting HIV sites of vulnerability

针对艾滋病毒易感部位的疫苗

• 批准号: 10512063
• 负责人: Catarina E Hioe
• 金额: --
• 依托单位: JAMES J PETERS VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10512063
• 关键词: 2019-nCoV; AIDS prevention; Adherence; Anti-Retroviral Agents; Antibodies; Antibody Formation; Antibody Response; Antigens; Biomanufacturing; COVID-19; Caring; Cells; Clinical Trials; Collaborations; Complement Activation; DNA; Data; Development; Disease; Epitopes; Escherichia coli; Future; Glycoproteins; Goals; Good Manufacturing Process; HIV; HIV Envelope Protein gp120; HIV Infections; HIV envelope protein; HIV vaccine; Healthcare Systems; Home; Human; Immune; Immune response; Immunization; Immunize; Immunodominant Epitopes; Immunoglobulin G; In Vitro; Individual; Infection; Laboratories; Lead; Legal patent; Macaca mulatta; Mediating; Medical center; Methods; Monkeys; Monoclonal Antibodies; Oryctolagus cuniculus; Outcome; Pathway interactions; Phase; Phase I Clinical Trials; Plasma; Plasmids; Production; Protein Analysis; Proteins; Provider; Publishing; RNA Splicing; Recombinants; Regimen; Research; Risk; SIV; Scaffolding Protein; Site; Supervision; Testing; Transfection; United States Department of Veterans Affairs; United States Dept. of Health and Human Services; Universities; Vaccination; Vaccine Clinical Trial; Vaccines; Vagina; Validation; Veterans; Veterans Health Administration; Viral Physiology; Virion; Virus; Virus Diseases; Wisconsin; antibody-dependent cell cytotoxicity; antibody-dependent cellular phagocytosis; cross reactivity; design; economic impact; efficacy clinical trial; env Gene Products; glycosylation; immunogenic; immunogenicity; improved; in vivo; medical schools; monomer; nonhuman primate; novel; novel vaccines; pandemic disease; pathogen; plasmid DNA; prevent; protein purification; prototype; rational design; scaffold; simian human immunodeficiency virus; standard of care; vaccine candidate; vaccine efficacy; vaccine strategy; vaccine trial; vaccine-induced antibodies; virus envelope

Project Summary/Abstract Effective HIV vaccines are not yet available. Of the phase 2b/3 vaccine trials, only the Thai RV144 trial showed efficacy (31%, p=0.04), and high levels of antibodies (Abs) against the V1V2 domain of HIV envelope (Env) were found to be the only primary immune correlate of reduced virus acquisition. Studies of vaccines in SIV or SHIV-challenged monkeys have since recapitulated these findings. To improve upon the RV144 vaccine, we have designed V1V2-targeted vaccine candidates and identified the most immunogenic: V1V2/A244-2J9C, a protein with V1V2 of CRF_01.AE strain A244 spliced into a bacterial trimeric protein scaffold, 2J9C. With our unique vaccine strategy centered on targeting V1V2 using novel recombinant subunit immunogens, we have demonstrated the capacity to induce an Ab response in plasma and vaginal secretion that was focused on V1V2 and diverted from other more immunodominant sites on Env. The elicited Abs displayed cross-reactivity with strains from multiple clades, durability of 1-2 years after the last boost, and antiviral functions including Ab- dependent cellular phagocytosis (ADCP), Ab-dependent cellular cytotoxicity (ADCC), and complement activation—activities that are not readily achieved by immunization with intact gp120. V1V2/A244-2J9C DNA has also been shown as an effective prime, focusing the Ab response on the specific V2 region that accounted for reduced infection in RV144. Altogether these data provide evidence supporting the validation of our lead vaccine candidate and vaccination approach. A US patent has been issued for our V1V2-scaffold designs, with the VA as one of the assignees. This application proposes to produce the lead immunogen V1V2/A244-2J9C under cGMP (current Good Manufacturing Practice) as both DNA plasmid and protein, and test its optimized delivery in order to pave the way toward a human phase I clinical trial. To accomplish this goal, four specific aims are proposed. Aim 1 is to generate a master bank of E. coli transformed with the V1V2/A244-2J9C-expressing DNA plasmid. In Aim 2 we will test the cGMP-grade V1V2/A244-2J9C-encoding DNA for protein production in transiently transfected 293T cells and for immunogenicity in rabbits. In vitro protein analysis will include expression efficiency, mass, oligomerization, glycosylation, stability, and reactivity with a panel of monoclonals Abs (mAbs). Aim 3 is to produce a master bank of HEK293T GnTi-/- cells and a pilot batch of V1V2/A244-2J9C protein using the plasmid from Aim 1. Finally, Aim 4 will test purification methods and conduct in vitro analysis for V1V2/A244-2J9C protein from Aim 3 and then perform in vivo rabbit immunogenicity testing with V1V2/A244-2J9C DNA and protein. The cGMP production will be done by Waisman BioManufacturing, associated with the University of Wisconsin-Madison, under the supervision of Drs. Carl A. Ross and Brian M. Dattilo. In vitro immunogen analysis, protein purification, rabbit vaccination, and immune assessment will be performed in the laboratory of Dr. Catarina Hioe (PI, James J. Peters VA Medical Center, JJP VAMC) in collaboration with Dr. Susan Zolla-Pazner (Mount Sinai School of Medicine, MSSM). The V1V2/A244-2J9C DNA and protein immunogens will be the first of their kind to move toward clinical trials. An effective vaccine to prevent HIV infection and/or disease is an essential portion of the Strategic National Vaccine Plan of the Departments of Health and Human Services and Veterans Affairs. An HIV vaccine is invaluable to protect Veterans who are at risk at home and abroad. This vaccine could as well serve as a prototype for vaccines against other diseases, like COVID19, where a focused Ab response to specific epitopes is requisite for protection.

项目概要/摘要 目前还没有有效的艾滋病毒疫苗。在2b/3期疫苗试验中，只有泰国RV144试验 显示出功效（31%，p=0.04），并且针对 HIV 包膜 V1V2 结构域的抗体 (Abs) 水平很高 (Env)被发现是减少病毒获得的唯一主要免疫相关因素。疫苗研究 此后，SIV 或受到 SHIV 攻击的猴子也重复了这些发现。为了改进 RV144 疫苗， 我们设计了针对 V1V2 的候选疫苗并确定了最具免疫原性的疫苗：V1V2/A244-2J9C， 具有 CRF_01.AE 菌株 A244 的 V1V2 的蛋白质剪接到细菌三聚体蛋白质支架 2J9C 中。与我们的 以使用新型重组亚基免疫原针对 V1V2 为中心的独特疫苗策略，我们有 证明了在血浆和阴道分泌物中诱导针对 V1V2 的抗体反应的能力 并从 Env 上其他更具免疫优势的位点转移。引发的 Abs 显示出与 来自多个分支的菌株，最后一次加强后 1-2 年的持久性，以及包括 Ab- 在内的抗病毒功能 依赖性细胞吞噬作用 (ADCP)、抗体依赖性细胞毒性 (ADCC) 和补体 激活——用完整的 gp120 免疫不容易实现的活性。 V1V2/A244-2J9C DNA 有 也被证明是一种有效的引物，将抗体反应集中在特定的 V2 区域上，从而解释了 RV144 感染减少。总而言之，这些数据提供了支持我们的先导疫苗验证的证据 候选人和疫苗接种方法。我们的 V1V2 支架设计已获得美国专利，VA 作为受让人之一。 本申请拟在cGMP（现行良好生产规范）下生产先导免疫原V1V2/A244-2J9C 制造实践）作为DNA质粒和蛋白质，并测试其优化的交付，以铺平 迈向人体 I 期临床试验的道路。为了实现这一目标，提出了四个具体目标。目标 1 是 生成用 V1V2/A244-2J9C 表达 DNA 质粒转化的大肠杆菌主库。在目标 2 中，我们 将测试 cGMP 级 V1V2/A244-2J9C 编码 DNA 在瞬时转染的 293T 中的蛋白质生产 细胞和兔子的免疫原性。体外蛋白质分析将包括表达效率、质量、 寡聚化、糖基化、稳定性以及与一组单克隆抗体 (mAb) 的反应性。目标 3 是 使用该质粒产生 HEK293T GnTi-/- 细胞的主库和 V1V2/A244-2J9C 蛋白的中试批次 来自目标1。最后，目标4将测试纯化方法并对V1V2/A244-2J9C蛋白进行体外分析 来自目标 3，然后使用 V1V2/A244-2J9C DNA 和蛋白质进行体内兔免疫原性测试。 cGMP 生产将由与威斯曼大学合作的 Waisman BioManufacturing 完成 威斯康星-麦迪逊，在博士的监督下。卡尔·A·罗斯和布莱恩·M·达蒂洛。体外免疫原分析， 蛋白质纯化、兔子疫苗接种和免疫评估将在 Dr. 的实验室进行。 Catarina Hioe（PI、James J. Peters VA 医疗中心、JJP VAMC）与 Susan Zolla-Pazner 博士合作 （西奈山医学院，MSSM）。 V1V2/A244-2J9C DNA和蛋白质免疫原将是同类中第一个走向临床的免疫原 试验。预防艾滋病毒感染和/或疾病的有效疫苗是国家战略的重要组成部分 卫生与公众服务部和退伍军人事务部的疫苗计划。 HIV 疫苗是 对于保护国内外处于危险中的退伍军人来说是无价的。这种疫苗也可以作为 针对其他疾病（如 COVID19）的疫苗原型，其中针对特定表位的集中抗体反应 是保护所必需的。