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: 10692084
• 负责人: Ursula Buchholz
• 金额: $ 22.19万
• 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/10692084
• 关键词: Adult; Adverse event; Aerosols; Alphavirus; Animals; Antibodies; Antibody Response; Antibody titer measurement; Antigens; Attenuated; Avian Influenza A Virus; Avulavirus; Biological Assay; Birds; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; COVID-19 pandemic; Cattle; Cavia; Cell Culture Techniques; Cells; Child; Chimera organism; Clinical; Clinical Research; Coronavirus spike protein; Development; Disease; 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; Membrane Glycoproteins; Membrane Proteins; Messenger RNA; Modification; Mucosal Immunity; Mucous Membrane; 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; Serum; Severe Acute Respiratory Syndrome; Signal Transduction; Surface Antigens; System; TRIP10 gene; Testing; Time; United States National Institutes of Health; Upper respiratory tract; Vaccination; Vaccines; Viral; Viral Vaccines; Virion; Virus; Virus Replication; Virus Shedding; Work; airway epithelium; antigen test; attenuation; base; cohort; conjunctiva; design; emerging pathogen; expression vector; immunogenic; immunogenicity; mucosal vaccine; neutralizing antibody; nonhuman primate; open label; parainfluenza virus; pathogen; pathogenic virus; phase 1 study; respiratory; respiratory pathogen; 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 had also 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 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 also 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. We have initiated a Phase 1 study to evaluate the safety, infectivity, and immunogenicity of this second-generation HPIV3/delHNF/EbovZ GP vaccine candidate when administered intranasally in healthy adults in an inpatient setting (NCT03462004). This vaccine candidate contains EbovZ GP as the sole envelope glycoprotein. 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/delHNF/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 SARS-CoV-2 pandemic will be lifted. Participants in Cohort 2 will be randomly assigned to receive two doses of either 7.0 log10 PFU/mL of HPIV3/delHNF/EbovZ-GP vaccine or placebo on Days 0 and 28. We recently collaborated with Alexander Bukreyev and colleagues (UTMB Galveston, TX) to compare the qualitative and quantitative humoral parameters of different intranasal vector vaccines expressing EbovZ-GP in cynomolgus macaques. Of the five Ebov GPexpressing mucosal vaccines derived from human and avian paramyxoviral vectors (based on versions of HPIV1, HPIV3, or Newcastle disease virus), the HPIV3/delHNF/EbovZ GP vaccine conferred NHPs with the best protection, leaving them free of disease with near-sterilizing immunity against Ebov. Thus, the continued evaluation of the HPIV3/delHNF/EbovZ GP vaccine candidate in the Phase 1 study (described above) is warranted.

我们以前构建了一种称为HPIV3-EBOVZ GP的第一代构建体，其中HPIV3的JS菌株的完整基因组通过在HPIV3 P和M基因之间的第三个基因位置中添加在第三个基因位置中修改了HPIV3的完整基因组。基于先前的临床研究，认为JS菌株被认为是衰减的HPIV3，尽管这种衰减的基础尚不清楚。 EBOV GP是唯一的EBOV病毒表面蛋白，唯一的EBOV中和抗原和主要的保护性抗原。 EBOV GP基因经过设计，具有适当的HPIV3转录信号，以通过HPIV3聚合酶表示为单独的mRNA。当通过鼻内（IN）和气管内（IT）给药给予非人类灵长类动物时，HPIV3-EBOVZ GP即使在先前感染了HPIV3感染的动物中，都具有免疫原性和保护性。但是，免疫原性取决于IT疫苗的输送：仅输送不足。这表明超出上呼吸道的载体表达对于免疫原性是必需的。我们还探索了恒河猕猴中的气溶胶路线的HPIV3-EBOVZ GP结构。与/IT路线中的组合相比，气溶胶途径通常更具免疫原性和保护性。这通常诱导了较高的血清和粘膜EBOV特异性IgG，IgA和中和抗体滴度，以及肺中的EBOV特异性细胞反应，包括多功能的CD8+ T细胞和CD4+ T Helper细胞，主要是Th1。此外，HPIV3-EBOVZ GP疫苗诱导的细胞介导的和体液免疫反应比平行于养生的α疫苗更强大。 HPIV3-EBOVZ GP的一种气溶胶剂量赋予了100％保护猕猴，以防止EBOV挑战。 我们进行了（约翰·霍普金斯彭博公共卫生学院的临床合作者）一项开放标签的1阶段临床试验，以确定HPIV3-EBOVZ GP的安全性，耐受性和免疫原性在健康成年人中提供的健康成年人在患者中的设置（NCT025645750），以前是一项安全研究，以前是安全性的。十名受试者接受了两剂（4至8周间隔）为6.0 Log10 PFU的疫苗。第一个剂量是中等感染的（7/10受试者脱离了QRT-PCR检测到的病毒，平均峰值滴度3.8 Log10基因组等效物/mL，平均脱落的持续时间为7.9天）。第二剂剂量后发现很少的脱落。第二个队列（n = 20）接受了两种计划的7.0 log10 PFU疫苗的剂量之一。脱落相似，但持续时间较短（平均为3.7天）。疫苗的耐受性良好，除了在疫苗接种后在队列2中发现5名志愿者（3个温和，2个中度）的无症状ALT升高，并且与脱落有关。所有这些都在第28天解决了。由于这些Alts的这些升高，该研究已停止，但其意义尚不清楚。因此，目前将不进一步施用该疫苗。血清抗体的诱导很差（粘膜抗体尚未分析），但是这是预期的，如上所述，我们先前已经观察到，仅在恒河猴中，仅通过途径给药是免疫原性的。 我们还开发了该向量的第二代版本，称为HPIV3/delhnf/eBovz-GP，其中HPIV3 F和HN基因被删除，使Ebov GP成为唯一的病毒表面糖蛋白。 我们已经开始了一项1阶段研究，以评估这种第二代HPIV3/delhnf/ebovz GP GP疫苗候选者的安全性，感染性和免疫原性，当时在健康的成年人中内鼻内给药（NCT03462004）。该疫苗候选者包含Ebovz GP作为唯一的包膜糖蛋白。参与者在两个队列中被依次入学。队列1的参与者已被随机分配，以接收两剂6.0 log10 pfu/ml的HPIV3/delhnf/ebovz-GP疫苗或安慰剂。第一个剂量在第0天服用，第二剂剂量在35天后给予。通过鼻腔清洗和RT-QPCR评估疫苗复制和感染性测定，并将测量血清抗体检测。不出所料，在6.0 log10 PFU剂量下，HPIV3/delhnf/ebovz-GP疫苗的感染率很小，不良事件通常是轻度至中度的。该研究被认为可以安全地进行评估，以评估由于SARS-COV-2大流行而导致的闭合后，将进行较高的7.0 log10 pfu剂量。队列2的参与者将随机分配，以在第0和28天接受两剂HPIV3/delhnf/ebovz-GP疫苗或安慰剂的HPIV3/delhnf/ebovz-GP疫苗或安慰剂。 我们最近与亚历山大·布克里耶夫（Alexander Bukreyev）及其同事（德克萨斯州加尔维斯顿（UTMB Galveston））合作，比较了在cynomolgus猕猴中表达eBovz-gp的不同鼻内矢量疫苗的定性和定量体液参数。在五个EBOV表达粘膜疫苗中，源自人和禽类丙糖病毒载体（基于HPIV1，HPIV3或纽卡斯尔病毒的版本），HPIV3/delhnf/ebovz GP GP疫苗，使其与近距离化的ebov无效，使其无效地保护疾病。因此，必须对HPIV3/DELHNF/EBOVZ GP疫苗在第一阶段研究中的持续评估（如上所述）。