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.

我们先前构建了称为HPIV 3-EbovZ GP的第一代构建体，其中通过在HPIV 3 P和M基因之间的第三基因位置添加EBOV GP基因来修饰HPIV 3的JS株的完整基因组。根据先前的临床研究，JS毒株被认为是减毒的HPIV 3，尽管这种减毒的基础尚不清楚。EBOV GP是唯一的EBOV病毒粒子表面蛋白、唯一的EBOV中和抗原和主要的保护性抗原。EBOV GP基因被工程化以具有适当的HPIV 3转录信号，用于通过HPIV 3聚合酶将其表达为单独的mRNA。当通过鼻内（IN）和气管内（IT）联合给药给予非人灵长类动物时，HPIV 3-EbovZ GP具有显著的免疫原性和保护性，即使在先前感染HPIV 3的动物中也是如此。然而，免疫原性依赖于疫苗的IT递送：单独的IN递送是不够的。这表明载体在上呼吸道以外的表达是免疫原性所必需的。 因此，我们探索了在恒河猴中通过气溶胶途径递送HPIV 3-EbovZ GP构建体。气雾剂途径通常比IN/IT联合途径更具免疫原性和保护性。这通常诱导更高的血清和粘膜EBOV特异性IgG、伊加和中和抗体滴度，以及肺中的EBOV特异性细胞应答，包括多功能CD 8 + T细胞和主要是Th 1的CD 4 + T辅助细胞。此外，HPIV 3-EbovZ GP疫苗诱导的细胞介导的和体液免疫应答比肠胃外平行递送的甲病毒疫苗更稳健。一个气雾剂剂量的HPIV 3-EbovZ GP赋予猕猴100%的保护以抵抗EBOV攻击。 我们开发了该载体的第二代版本，称为HPIV 3/delHNF/EBOV Z-GP，其中HPIV 3 F和HN基因被删除，留下EBOV GP作为唯一的病毒表面糖蛋白。我们的合作者亚历山大Bukreyev在得克萨斯大学医学分支，加尔维斯顿的一项大型比较研究中食蟹猴（他在LID/NIAID的工作人员科学家时制作了该构建体）显示，这种第二代版本甚至比第一代版本更具保护性，即使它的复制受到非常高度的限制（比第一代构建体受到的限制要多得多）。 我们（与约翰霍普金斯彭博公共卫生学院的临床合作者）进行了一项开放标签I期临床试验，以确定在住院环境中IN递送的HPIV 3-EbovZ GP在健康成人中的安全性、耐受性和免疫原性（NCT 025645750），该试验旨在作为评价气雾剂递送之前的安全性研究。10例受试者接受了两剂（间隔4- 8周）6.0 log 10 PFU的疫苗。首次给药具有中度感染性（通过qRT-PCR检测到7/10例受试者排毒，平均峰值滴度3.8 log 10基因组当量/ml，平均排毒持续时间7.9天）。第二次给药后检测到少量散粒。第二个队列（n=20）接受了两次计划剂量7.0 log 10 PFU疫苗中的一次。脱落相似，但持续时间较短（平均3.7天）。疫苗耐受性良好，但队列2的5名志愿者（3名轻度，2名中度）在接种疫苗后观察到无症状ALT升高，并与散毒相关。第28天全部消退。这项研究由于ALT的升高而停止，但其意义尚不清楚。因此，此时将不会进一步接种该疫苗。血清抗体的诱导较差（尚未分析粘膜抗体），但这是预期的，因为如上所述，我们之前观察到恒河猴中单独通过IN途径给药的免疫原性较差。 我们已经启动了一项I期研究，以评价住院健康成人鼻内接种两剂HPIV 3/HNF/EbovZ GP候选疫苗的安全性、感染性和免疫原性（NCT 03462004）。受试者按顺序入组两个队列。队列1中的参与者被随机分配接受两剂6.0 log 10 PFU/mL的HPIV 3/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。通过鼻洗液和RT-qPCR和感染性试验评价疫苗复制，并测量血清抗体应答。正如预期，在6.0 log 10 PFU剂量下，HPIV 3/HNF/EbovZ-GP疫苗具有轻微感染性，不良事件通常为轻度至中度。在因当前SARS-CoV-2大流行而关闭的研究将被解除后，该研究被认为是安全的，可以继续评价更高的7.0 log 10 PFU剂量。队列2中的受试者将被随机分配在第0天和第28天接受两剂7.0 log 10 PFU/mL HPIV 3/HNF/EbovZ-GP疫苗或安慰剂。