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Principal Investigator: Ursula Buchholz
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2021
Award: $215,100
Funding agency: National Institute of Allergy and Infectious Diseases
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 log10PFU/mL of HPIV3/delHNF/EbovZ-GP vaccine or placebo on Days 0 and 28.
We contributed to a study by Alexander Bukreyev and colleagues (UTMB Galveston, TX) that evaluated and compared the qualitative and quantitative humoral parameters of different intranasal vector vaccines expressing EbovZ-GP in cynomolgus macaques. Correlation network analysis defined RBD-specific anti- bodies and Fc-mediated immune functions as contributing factors to improved survival after Ebola virus challenge. Binding and neutralizing titers, isotype composition, total sera binding to GP as determined by biolayer inferometry, the proportion of binding and neutralizing antibodies against certain GP regions, linear epitope recognition patterns, and Fc-mediated effects were among some of the qualitative and quantitative response features analyzed. Pairwise network correlation analysis of 139 immune- and vaccine-related parameters was performed to demonstrate relationships with survival. Total GP-specific antibodies, as measured by biolayer interferometry, but not neutralizing IgG or IgA titers, correlated with survival. Fc-mediated functions and the amount of receptor binding domain antibodies were associated with improved survival outcomes, alluding to the protective mechanisms of these vaccines. Therefore, functional qualities of the antibody response, particularly Fc-mediated effects and GP specificity, rather than simply magnitude of the response, appear central to vaccine- induced protection against Ebov. 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.
Terms: <0-11 years old><21+ years old><7S Gamma Globulin><Adult><Adult Human><Adverse Experience><Adverse event><Aerosols><Alpha Virus><Alphavirus><Animals><Antibodies><Antibody Response><Antibody titer measurement><Antigenic Determinants><Antigens><Assay><Attenuated><Aves><Avian><Avian Influenza A Virus><Avian Influenza Virus><Avian Orthomyxovirus Type A><Avian Paramyxovirus 1><Avian Paramyxoviruses><Avulavirus><Binding><Binding Determinants><Bioassay><Biologic Assays><Biological Assay><Birds><Blood Serum><Bovine Species><CD4 Cells><CD4 Positive T Lymphocytes><CD4 T cells><CD4 helper T cell><CD4 lymphocyte><CD4+ T-Lymphocyte><CD4-Positive Lymphocytes><CD8 Cell><CD8 T cells><CD8 lymphocyte><CD8+ T cell><CD8+ T-Lymphocyte><CD8-Positive Lymphocytes><CD8-Positive T-Lymphocytes><CDC42 Homolog Gene><CDC42-Interacting Protein Gene><CIP4><COVID crisis><COVID epidemic><COVID pandemic><COVID-19 crisis><COVID-19 epidemic><COVID-19 global health crisis><COVID-19 global pandemic><COVID-19 health 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Virus><G25K Gene><GP Ebola virus><GP Gene><Gene Arrangement><Gene Order><Gene Position><Gene Transcription><Generations><Genes><Genetic Transcription><Genome><Genomics><Glycoproteins><Group A Arboviruses><Guinea Pigs><Guinea Pigs Mammals><HPIV1><HPIV3><Hemadsorption Type 2 Virus><Hemadsorption Virus 1><Hemadsorption Virus 2><Human><Human Parainfluenza Virus 1><Human Parainfluenza Virus 3><IgA><IgG><Immune><Immune response><Immunes><Immunity><Immunoglobulin A><Immunoglobulin G><Immunologic Surface Markers><Immunological Surface Markers><Immunological response><Infant><Infection><Influenza HA><Influenza Hemagglutinin><Inpatients><Interferometry><Intracellular Communication and Signaling><Intranasal Administration><Intranasal Drug Administration><Lifting><Lung><Lung Respiratory System><M fascicularis><M mulatta><M. fascicularis><M. mulatta><Macaca><Macaca fascicularis><Macaca mulatta><Macaque><Measures><Mediating><Membrane Glycoproteins><Membrane Protein Gene><Membrane 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Acid><Rodent><Rodentia><Rodents Mammals><Route><SARS><SARS Virus><SARS corona virus><SARS coronavirus><SARS coronavirus disease><SARS-Associated Coronavirus><SARS-CoV><SARS-CoV disease><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 pandemic><SARS-CoV2 epidemic><SARS-CoV2 pandemic><SARS-Related Coronavirus><SARS-coronavirus-2 epidemic><SARS-coronavirus-2 pandemic><Safety><Serum><Severe Acute Respiratory Syndrome><Severe Acute Respiratory Syndrome CoV 2 epidemic><Severe Acute Respiratory Syndrome CoV 2 pandemic><Severe Acute Respiratory Syndrome CoV disease><Severe Acute Respiratory Syndrome Virus><Severe Acute Respiratory Syndrome corona virus><Severe Acute Respiratory Syndrome coronavirus><Severe Acute Respiratory Syndrome coronavirus disease><Severe acute respiratory syndrome coronavirus 2 epidemic><Severe acute respiratory syndrome coronavirus 2 pandemic><Sham Treatment><Signal Transduction><Signal Transduction 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