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: 10927793
• 负责人: Ursula Buchholz
• 金额: $ 10.76万
• 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/10927793
• 关键词: Adult; Adverse event; Aerosols; Alphavirus; Animal Experiments; 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; Cattle; Cavia; Cell Culture Techniques; Cells; Child; Chimera organism; Clinical; Clinical Research; Collaborations; Coronavirus spike protein; Cytoplasm; Development; Disease; Dose; Ebola virus; Ebola virus envelope glycoprotein; Effectiveness; Engineering; Enrollment; Evaluation; Family member; Gene Order; Generations; Genes; Genetic Transcription; Genome; Genomics; Glycoproteins; Helper-Inducer T-Lymphocyte; Human; Immune response; Immunity; Immunoglobulin A; Immunoglobulin G; Infant; Infection; Influenza Hemagglutinin; Inpatients; Intranasal Administration; Lung; Macaca; Macaca fascicularis; Macaca mulatta; Measures; Mediating; Membrane Glycoproteins; Membrane Proteins; Messenger RNA; Modification; Mucosal Immunity; Mucous Membrane; Nasal Lavage Fluid; Newcastle disease virus; 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; 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; viral detection; 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 to be 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. 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.

我们之前构建了称为 HPIV3-EbovZ GP 的第一代构建体，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 基因来修饰 HPIV3 JS 株的完整基因组。根据之前的临床研究，JS 株被认为是 HPIV3 减毒株，但这种减毒的基础尚不清楚。 Ebov GP是唯一的Ebov病毒颗粒表面蛋白、唯一的Ebov中和抗原和主要的保护性抗原。 Ebov GP 基因被设计为具有适当的 HPIV3 转录信号，以便通过 HPIV3 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，HPIV3-EbovZ GP 具有显着的免疫原性和保护性，甚至在先前感染 HPIV3 的动物中也是如此。然而，免疫原性取决于疫苗的 IT 递送：仅 IN 递送是不够的。这表明上呼吸道以外的载体表达对于免疫原性是必要的。我们还探索了在恒河猴中通过气雾剂途径递送 HPIV3-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。这通常会诱导较高的血清和粘膜 EBOV 特异性 IgG、IgA 和中和抗体滴度，以及肺部的 Ebov 特异性细胞反应，包括主要为 Th1 的多功能 CD8+ T 细胞和 CD4+ T 辅助细胞。此外，HPIV3-EbovZ GP 疫苗比并行注射的甲病毒疫苗诱导更强烈的细胞介导和体液免疫反应。一剂 HPIV3-EbovZ GP 气雾剂可为猕猴提供 100% 的保护，使其免受 EBOV 攻击。 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 1 期临床试验，以确定在住院环境中健康成人中注射 HPIV3-EbovZ GP 的安全性、耐受性和免疫原性 (NCT025645750)，该试验旨在作为评估气雾剂输送之前的安全性研究。 10 名受试者接受了两剂 6.0 log10 PFU 疫苗（间隔 4 至 8 周）。第一剂具有中等传染性（通过 qRT-PCR 检测到 7/10 的受试者排出了病毒，平均峰值滴度为 3.8 log10 基因组当量/ml，平均排出持续时间为 7.9 天）。第二次给药后几乎没有检测到脱落。第二组 (n=20) 接受了两剂 7.0 log10 PFU 疫苗的计划剂量之一。脱落情况相似，但持续时间较短（平均 3.7 天）。该疫苗耐受性良好，但第 2 组中的 5 名志愿者（3 名轻度，2 名中度）在接种疫苗后发现无症状 ALT 升高，且与排毒相关。所有问题均在第 28 天解决。该研究因 ALT 升高而停止，但其意义尚不清楚。因此，目前不会进一步接种该疫苗。血清抗体的诱导较差（粘膜抗体待分析），但这是预料之中的，因为如上所述，我们之前观察到单独通过IN途径给药在恒河猴中免疫原性较差。 我们还开发了该载体的第二代版本，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。 我们已经启动了一项 1 期研究，以评估第二代 HPIV3/delHNF/EbovZ GP 候选疫苗在住院健康成人鼻内给药时的安全性、传染性和免疫原性 (NCT03462004)。该候选疫苗含有 EbovZ GP 作为唯一的包膜糖蛋白。参与者按顺序分为两个队列。第 1 组的参与者被随机分配接受两剂 6.0 log10 PFU/mL HPIV3/delHNF/EbovZ-GP 疫苗或安慰剂。第一剂在第 0 天注射，第二剂在 35 天后注射。通过鼻洗、RT-qPCR 和感染性测定评估疫苗复制，并测量血清抗体反应。正如预期的那样，在 6.0 log10 PFU 剂量下，HPIV3/delHNF/EbovZ-GP 疫苗具有轻微的传染性，不良事件通常为轻度至中度。该研究被认为可以安全地继续评估较高的 7.0 log10 PFU 剂量。第 2 组的参与者将被随机分配在第 0 天和第 28 天接受两剂 7.0 log10 PFU/mL HPIV3/delHNF/EbovZ-GP 疫苗或安慰剂。 我们最近与 Alexander Bukreyev 及其同事（德克萨斯州 UTMB 加尔维斯顿）合作，比较了食蟹猴中表达 EbovZ-GP 的不同鼻内载体疫苗的定性和定量体液参数。在源自人类和禽类副粘病毒载体（基于 HPIV1、HPIV3 或新城疫病毒版本）的五种表达 Ebov GP 的粘膜疫苗中，HPIV3/delHNF/EbovZ GP 疫苗为 NHP 提供了最好的保护，使其免受疾病影响，并具有针对埃博夫病毒的近乎绝育的免疫力。因此，有必要在 1 期研究（如上所述）中继续评估 HPIV3/delHNF/EbovZ GP 候选疫苗。

项目成果

Respiratory tract immunization of non-human primates with a Newcastle disease virus-vectored vaccine candidate against Ebola virus elicits a neutralizing antibody response.

• DOI: 10.1016/j.vaccine.2010.10.024
• 发表时间: 2010-12-10
• 期刊: VACCINE
• 影响因子: 5.5
• 作者: DiNapoli, Joshua M.;Yang, Lijuan;Samal, Siba K.;Murphy, Brian R.;Collins, Peter L.;Bukreyev, Alexander
• 通讯作者: Bukreyev, Alexander

Comparative immunogenicity of HIV-1 gp160, gp140 and gp120 expressed by live attenuated newcastle disease virus vector.

• DOI: 10.1371/journal.pone.0078521
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Khattar SK;Samal S;LaBranche CC;Montefiori DC;Collins PL;Samal SK
• 通讯作者: Samal SK

Immunization of cattle with recombinant Newcastle disease virus expressing bovine herpesvirus-1 (BHV-1) glycoprotein D induces mucosal and serum antibody responses and provides partial protection against BHV-1.

• DOI: 10.1016/j.vaccine.2010.02.051
• 发表时间: 2010-04-19
• 期刊: VACCINE
• 影响因子: 5.5
• 作者: Khattar, Sunil K.;Collins, Peter L.;Samal, Siba K.
• 通讯作者: Samal, Siba K.

Complete genome sequence of avian paramyxovirus-3 strain Wisconsin: evidence for the existence of subgroups within the serotype.

• DOI: 10.1016/j.virusres.2009.12.015
• 发表时间: 2010-04
• 期刊: VIRUS RESEARCH
• 影响因子: 5
• 作者: Kumar, Sachin;Nayak, Baibaswata;Samuel, Arthur S.;Xiao, Sa;Collins, Peter L.;Samal, Siba K.
• 通讯作者: Samal, Siba K.

Replication, neurotropism, and pathogenicity of avian paramyxovirus serotypes 1-9 in chickens and ducks.

禽副粘病毒血清型 1-9 在鸡和鸭中的复制、向神经性和致病性。

• DOI: 10.1371/journal.pone.0034927
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Kim,Shin-Hee;Xiao,Sa;Shive,Heather;Collins,PeterL;Samal,SibaK
• 通讯作者: Samal,SibaK