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.

项目摘要/摘要 有效的HIV疫苗尚未可用。在2B/3期疫苗试验中，仅泰国RV144试验 显示效率（31％，p = 0.04）和针对HIV包膜V1V2域的高水平抗体（ABS） 发现（ENV）是降低病毒获取的唯一主要免疫相关性。疫苗的研究 此后，SIV或SHIV挑战的猴子概括了这些发现。为了改善RV144疫苗， 我们已经设计了V1V2靶向的疫苗候选物，并确定了最免疫原性：V1V2/A244-2J9C， 具有crf_01.ae菌株A244的V1V2的蛋白质剪接到三聚蛋白支架中的蛋白质，2J9C。与我们的 独特的疫苗策略以使用新型重组亚基免疫原为靶向V1V2的唯一疫苗策略，我们有 证明了在V1V2上诱导血浆和阴道分泌中AB反应的能力 并从Env上的其他更为免疫主导地点转移。引起的ABS显示与 来自多个进化枝的菌株，最后提升后1 - 2年的耐用性，以及包括AB-的抗病毒功能 依赖性细胞吞噬作用（ADCP），AB依赖性细胞毒性（ADCC）和完成 激活 - 通过完整的GP120免疫不容易实现的活性。 V1V2/A244-2J9C DNA具有 还显示为有效的素数，将AB响应集中在考虑的特定V2区域上 RV144中的感染降低。这些数据总共提供了支持我们铅疫苗验证的证据 候选和疫苗接种方法。已通过VA颁发了美国V1V2-SCASTADGOLD设计的美国专利 作为指派之一。 该申请提案在CGMP下生产铅免疫原V1V2/A244-2J9C（电流良好 制造实践）作为DNA质粒和蛋白质，并测试其优化的递送以铺平 进入人类I期临床试验的方式。为了实现这一目标，提出了四个具体目标。目标1是 通过V1V2/A244-2J9C表达DNA质粒的大肠杆菌产生大肠杆菌的主库。在目标2中我们 将测试CGMP级V1V2/A244-2J9C编码DNA的蛋白质生产，以瞬时翻译为293T 细胞和兔子的免疫原性。体外蛋白分析将包括表达效率，质量， 寡聚，糖基化，稳定性和反应性与一组单克隆ABS（mAb）。目标3是 使用质粒生产HEK293T GNTI - / - 细胞的HEK293T GNTI - / - 细胞和一批V1V2/A244-2J9C蛋白 从AIM 1。最后，AIM 4将测试纯化方法并进行V1V2/A244-2J9C蛋白的体外分析 从AIM 3中，然后使用V1V2/A244-2J9C DNA和蛋白质进行体内兔免疫原性测试。 CGMP的生产将由Waisman Biomenturativer进行，与大学有关 威斯康星州麦迪逊，在博士的监督下。卡尔·罗斯和布莱恩·达蒂洛。体外免疫分析， 蛋白质纯化，兔疫苗接种和免疫评估将在博士的实验室中进行。 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和蛋白质免疫原子将是第一个朝着临床迈进的一种 试验。预防艾滋病毒感染和/或疾病的有效疫苗是战略国家的重要部分 卫生与公共服务部和退伍军人事务部门的疫苗计划。 HIV疫苗是 保护在国内外有危险的退伍军人的宝贵。这种疫苗也可以用作 针对其他疾病（例如Covid19）的疫苗的原型，其中集中对特定表位的AB反应 是保护的必要条件。