Paramyxoviruses as Vaccine Vectors Against Highly Pathogenic Viruses

副粘病毒作为高致病性病毒的疫苗载体

• 批准号: 10272101
• 负责人: Ursula Buchholz
• 金额: $ 27.96万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10272101
• 关键词: 2019-nCoV; Adult; Adverse event; Aerosols; Alphavirus; Animals; Antibodies; Antibody Response; Antibody titer measurement; Antigens; Attenuated; Avian Influenza A Virus; Avulavirus; Biological Assay; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cattle; Cavia; Cell Culture Techniques; Cells; Child; Chimera organism; Clinical; Clinical Research; Comparative Study; Coronavirus spike protein; Development; Dose; Ebola virus; Ebola virus envelope glycoprotein; Effectiveness; Engineering; Enrollment; Evaluation; Family member; Gene Order; Generations; Genes; Genetic Transcription; Genome; Genomics; Glycoproteins; Human; Immune response; Immunity; Immunoglobulin A; Immunoglobulin G; Infant; Infection; Influenza Hemagglutinin; Inpatients; Intranasal Administration; Lifting; Lung; Macaca; Macaca fascicularis; Macaca mulatta; Measures; Mediating; Medical; Membrane Glycoproteins; Membrane Proteins; Messenger RNA; Modification; Mucosal Immunity; Mucous Membrane; National Institute of Allergy and Infectious Disease; Newcastle disease virus; Nose; Para-Influenza Virus Type 1; Para-Influenza Virus Type 3; Paramyxovirus; Participant; Pathogenicity; Phase I Clinical Trials; Placebos; Polymerase; Population; Primates; Proteins; Public Health Schools; Quantitative Reverse Transcriptase PCR; RNA; Randomized; Respiratory System; Rodent; Route; SARS coronavirus; Safety; Scientist; Serotyping; Serum; Severe Acute Respiratory Syndrome; Signal Transduction; Structure of respiratory epithelium; Surface Antigens; System; TRIP10 gene; Testing; Texas; Time; United States National Institutes of Health; Universities; Upper respiratory tract; Vaccination; Vaccines; Viral; Viral Vaccines; Virion; Virus; Virus Replication; Virus Shedding; Work; attenuation; base; cohort; conjunctiva; design; expression vector; immunogenic; immunogenicity; neutralizing antibody; nonhuman primate; open label; pandemic disease; parainfluenza virus; pathogen; pathogenic virus; phase 1 study; respiratory; response; reverse genetics; safety study; seropositive; tissue tropism; vaccine candidate; vaccine delivery; vaccine development; vector; vector vaccine; vector-based vaccine; volunteer

We previously constructed a first-generation construct called HPIV3-EbovZ GP, in which the complete genome of the JS strain of HPIV3 was modified by the addition of the EBOV GP gene in the third gene position, between the HPIV3 P and M genes. The JS strain is thought to be an attenuated HPIV3, based on previous clinical studies, although the basis of this attenuation is unknown. EBOV GP is the sole EBOV virion surface protein, the sole EBOV neutralization antigen, and the major protective antigen. The EBOV GP gene was engineered to have the appropriate HPIV3 transcription signals for it to be expressed as a separate mRNA by the HPIV3 polymerase. HPIV3-EbovZ GP was substantially immunogenic and protective when given to non-human primates by combined intranasal (IN) and intratracheal (IT) administration, even in animals previously infected with HPIV3. However, immunogenicity depended on IT delivery of vaccine: IN delivery alone was insufficient. This suggested that vector expression beyond the upper respiratory tract was necessary for immunogenicity. We therefore explored delivery of the HPIV3-EbovZ GP construct by the aerosol route in rhesus macaques. The aerosol route was generally more immunogenic and protective than the combined IN/IT route. This induced generally higher serum and mucosal EBOV-specific IgG, IgA, and neutralizing antibody titers, as well as EBOV-specific cellular responses in the lungs, including polyfunctional CD8+ T cells and CD4+ T helper cells that were predominately Th1. In addition, the HPIV3-EbovZ GP vaccine induced more robust cell-mediated and humoral immune responses than an alphavirus vaccine delivered parenterally in parallel. One aerosol dose of HPIV3-EbovZ GP conferred 100% protection to macaques against EBOV challenge. We developed a second-generation version of this vector, called HPIV3/delHNF/EbovZ-GP, in which the HPIV3 F and HN genes were deleted, leaving EBOV GP as the sole viral surface glycoprotein. A large comparative study in cynomolgus monkeys by our collaborator Alexander Bukreyev at the University of Texas Medical Branch, Galveston, (who made the construct while a Staff Scientist in LID/NIAID) showed that this second-generation version was even more protective than the first-generation even though it was very highly restricted for replication (much more restricted than the first-generation construct). We performed (with clinical collaborators at the Johns Hopkins Bloomberg School of Public Health) an open label phase 1 clinical trial to determine the safety, tolerability, and immunogenicity of HPIV3-EbovZ GP delivered IN in healthy adults in an inpatient setting (NCT025645750), which was intended to be a safety study prior to evaluating aerosol delivery. Ten subjects received two doses (4- to 8-week interval) of 6.0 log10 PFU of vaccine. The first dose was moderately infectious (7/10 subjects shed virus detected by qRT-PCR, mean peak titer 3.8 log10 genomic equivalents/ml, mean duration of shedding 7.9 days). Little shedding was detected after the second dose. A second cohort (n=20) received one of two planned doses of 7.0 log10 PFU of vaccine. Shedding was similar but of shorter duration (mean of 3.7 days). The vaccine was well tolerated, with the exception that asymptomatic ALT elevations were noted in 5 volunteers (3 mild, 2 moderate) in cohort 2 after vaccination and associated with shedding. All resolved by day 28. The study was halted due to these elevations of ALTs, but their significance is unclear. Because of this, this vaccine will not be administered further at this time. Induction of serum antibodies was poor (mucosal antibodies not yet analyzed), but this was expected since, as noted above, we had previously observed that administration by the IN route alone was poorly immunogenic in rhesus monkeys. We have initiated a Phase 1 study to evaluate the safety, infectivity, and immunogenicity of two doses of the HPIV3/HNF/EbovZ GP vaccine candidate when administered intranasally in healthy adults in an inpatient setting (NCT03462004). Participants are being enrolled sequentially in two cohorts. Participants in Cohort 1 have been randomly assigned to receive two doses of either 6.0 log10 PFU/mL of HPIV3/delHNF/EbovZ-GP vaccine or placebo. The first dose was given on Day 0 and the second dose was given 35 days later. Vaccine replication was evaluated by nasal wash and RT-qPCR and infectivity assays, and serum antibody responses will be measured. As expected, at the 6.0 log10 PFU dose, the HPIV3/HNF/EbovZ-GP vaccine was marginally infectious, and adverse events were generally mild to moderate. The study was deemed safe to proceed to the evaluation of the higher 7.0 log10 PFU dose after the closures due to the current SARS-CoV-2 pandemic will be lifted. Participants in Cohort 2 will be randomly assigned to receive two doses of either 7.0 log10PFU/mL of HPIV3/HNF/EbovZ-GP vaccine or placebo on Days 0 and 28.

我们先前构建了一个称为HPIV3-EbovZ GP的第一代载体，其中通过在HPIV3 P和M基因之间的第三个基因位置添加EBOV GP基因来修改HPIV3 JS株的完整基因组。根据之前的临床研究，JS株被认为是一种减毒的HPIV3，尽管这种减毒的基础尚不清楚。EBOV gp是唯一的EBOV病毒表面蛋白、唯一的EBOV中和抗原和主要保护性抗原。EBOV gp基因被设计成具有适当的HPIV3转录信号，使其能够通过HPIV3聚合酶作为单独的mRNA表达。HPIV3-EbovZ GP通过鼻腔(IN)和气管(IT)联合给药给非人类灵长类动物，即使是在以前感染HPIV3的动物身上，也具有显著的免疫原性和保护性。然而，免疫原性依赖于疫苗的交付：仅交付疫苗是不够的。这表明载体在上呼吸道以外的表达对于免疫原性是必要的。 因此，我们探索了HPIV3-EbovZ GP通过气雾剂途径在恒河猴体内的传递。气雾剂途径通常比IN/IT联合途径更具免疫原性和保护性。这通常会导致血清和粘膜中EBOV特异性的Ig G、Ig A和中和抗体效价升高，以及肺部EBOV特异性的细胞反应，包括以Th1为主的多功能CD8+T细胞和CD4+T辅助细胞。此外，HPIV3-EbovZ GP疫苗比平行注射的甲型病毒疫苗诱导更强大的细胞介导和体液免疫反应。一次雾化剂量的HPIV3-EbovZ GP可100%保护猕猴抵抗EBOV攻击。 我们开发了该载体的第二代版本，称为HPIV3/delHNF/EbovZ-gp，其中删除了HPIV3F和HN基因，使EBOV gp成为唯一的病毒表面糖蛋白。加尔维斯顿德克萨斯大学医学分校的合作者Alexander Bukreyev(他是LID/NIAID的一名工作人员科学家)在食蟹猴中进行了一项大规模的比较研究，结果表明，第二代版本甚至比第一代版本更具保护性，尽管它对复制有非常严格的限制(比第一代结构要严格得多)。 我们(与约翰霍普金斯大学彭博公共卫生学院的临床合作者)进行了一项开放标签的第1阶段临床试验，以确定在住院环境中健康成年人注射的HPIV3-EbovZ GP的安全性、耐受性和免疫原性(NCT025645750)，这是一项在评估气雾剂释放之前的安全性研究。10名受试者接种了两剂(间隔4-8周)6.0log10pfu疫苗。第一次接种为中度感染(qRT-PCR检测到7/10的受试者脱落病毒，平均峰值滴度为3.8log10基因组当量/ml，平均脱落持续时间为7.9d)。第二次给药后几乎没有脱落。第二个队列(n=20)接受了两个计划剂量7.0log10pfu疫苗中的一个。脱落时间相似，但持续时间较短(平均3.7天)。该疫苗耐受性良好，除了在接种后队列2中有5名志愿者(3名轻度，2名中度)观察到与脱发相关的无症状ALT升高。所有问题都在第28天前解决了。由于高地海拔升高，这项研究暂停，但其意义尚不清楚。正因为如此，目前不会进一步接种这种疫苗。血清抗体的诱导性很差(粘膜抗体尚未分析)，但这是意料之中的，因为如上所述，我们之前观察到，仅通过IN途径给药对恒河猴的免疫原性很差。 我们已经启动了一项第一阶段研究，以评估两剂HPIV3/HNF/EbovZ GP候选疫苗在住院环境下对健康成年人进行鼻内接种时的安全性、传染性和免疫原性(NCT03462004)。参与者将按顺序分成两组进行登记。队列1的参与者被随机分配接受两剂剂量的HPIV3/delHNF/EbovZ-gp疫苗或安慰剂。第一剂在第0天给药，第二剂在35天后给药。通过鼻腔冲洗、RT-qPCR和传染性试验评估疫苗复制情况，并测定血清抗体反应。正如预期的那样，在6.0log10pfu剂量下，HPIV3/HNF/EbovZ-gp疫苗具有轻微的传染性，不良反应一般为轻度至中度。这项研究被认为是安全的，可以在当前SARS-CoV-2大流行导致的关闭将被解除后，继续评估7.0log10 PFU的较高剂量。队列2的参与者将被随机分配到第0天和第28天接受两剂剂量的HPIV3/HNF/EbovZ-gp疫苗或安慰剂。