Recombinant human/bovine parainfluenza virus 1 (HPIV1) expressing a chimeric RSV/HPIV1 F protein and uses thereof
Inventors
Collins, Peter • Liang, Bo • Munir, Shirin • Nutt, Anne Schaap • Buchholz, Ursula • Mackow, Natalie • Kwong, Peter • Graham, Barney • McLellan, Jason
Assignees
US Department of Health and Human Services
Publication Number
US-11591372-B2
Publication Date
2023-02-28
Expiration Date
2036-01-20
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Abstract
Recombinant paramyxoviruses including a viral genome encoding a heterologous gene are provided. In several embodiments, the recombinant paramyxovirus is a recombinant parainfluenza virus, such as a recombinant PIV3 including a viral genome encoding a heterologous respiratory syncytial virus F ectodomain linked to the transmembrane domain and the cytoplasmic tail of the F protein from the PIV3. Nucleic acid molecules including the genome of a recombinant paramyxoviruses are also provided. The recombinant viruses may advantageously be used in vaccine formulations, such as for vaccines against parainfluenza virus and respiratory syncytial virus.
Core Innovation
Recombinant paramyxoviruses including a viral genome encoding a heterologous gene are provided. In several embodiments, the recombinant paramyxovirus is a recombinant parainfluenza virus comprising a viral genome comprising a heterologous gene encoding a type I membrane protein with a recombinant RSV F ectodomain linked to a transmembrane domain and cytoplasmic tail of the paramyxovirus F protein. These recombinant paramyxoviruses may comprise viral genomes with the heterologous gene inserted between the genomic promoter and the gene encoding the N protein, or between the genes encoding the N and P proteins.
The problem being solved relates to paramyxoviruses causing many animal and human deaths annually, with human respiratory syncytial virus (RSV) and human parainfluenza viruses (HPIVs) being major causes of severe respiratory infections especially in infants and children worldwide. Despite the burden, development of effective vaccines against RSV and HPIV remains elusive. Previous vaccines expressing the RSV F protein in parainfluenza virus vectors had poor immunogenicity, partially due to inefficient incorporation of RSV F into vector particles, leading to weak immune responses and genetic instability.
The disclosed invention solves these issues by swapping the transmembrane domain and cytoplasmic tail of the RSV F protein with those of the paramyxovirus F protein, significantly increasing the incorporation of RSV F ectodomain into the envelope of recombinant paramyxoviruses. This leads to a multi-fold increase in expression on vector particles, dramatically improving elicitation of virus-neutralizing immune responses in vivo, including higher quality serum antibodies with potent neutralization activity. In addition, stabilization of the RSV F protein in the prefusion conformation further improves immunogenicity and protective efficacy. The invention also includes codon-optimization and insertion site optimization in the viral genome to maximize expression and reduce cytopathic effects like syncytium formation, enhancing safety and immunogenicity.
Claims Coverage
The independent claims disclose recombinant paramyxoviruses comprising genomes encoding RSV F ectodomains fused to paramyxovirus F protein transmembrane and cytoplasmic tail domains, plus their uses in immunogenic compositions and methods of eliciting immune responses.
Recombinant paramyxovirus expressing chimeric RSV F protein fused to paramyxovirus F TM and CT
A recombinant paramyxovirus comprising a viral genome encoding parainfluenza virus (PIV) N, P, M, F, HN, and L proteins, and a heterologous gene encoding a recombinant RSV fusion (F) ectodomain linked to the transmembrane domain and cytoplasmic tail of the PIV F protein; the recombinant paramyxovirus is a recombinant human parainfluenza virus 1 (HPIV1), wherein the heterologous gene comprises the nucleotide sequence set forth as SEQ ID NO: 136.
Insertion site and gene arrangement of heterologous RSV F gene in paramyxovirus genome
The heterologous gene encoding the recombinant RSV F ectodomain can be the first, second, or third gene downstream of a genomic promoter of the viral genome, positioned between the genomic promoter and the gene encoding the N protein or between the genes encoding the N and P proteins.
Attenuation mutations in recombinant HPIV1 vector backbones
The recombinant paramyxovirus may comprise a HPIV1 backbone encoding CΔ170 or LY942A attenuating mutations.
Efficient incorporation and envelope presentation of recombinant RSV F ectodomain
At least 90% of viral particles produced by a host cell infected with the recombinant paramyxovirus comprise a viral envelope containing the chimeric RSV F ectodomain.
Immunogenic compositions and methods of eliciting immune response
The recombinant paramyxovirus can be formulated into immunogenic compositions and used in methods for eliciting protective immune responses to RSV and/or PIV in subjects including intranasal administration.
The claims describe recombinant human parainfluenza virus 1 vectors comprising a heterologous gene encoding a recombinant RSV fusion (F) ectodomain fused to the PIV F protein transmembrane domain and cytoplasmic tail, with defined insertion sites and attenuation mutations, efficiently expressing RSV F on viral envelopes, formulated in immunogenic compositions useful for inducing immune responses to RSV and PIV.
Stated Advantages
Multi-fold increased incorporation of RSV F ectodomain into recombinant paramyxovirus envelope.
Dramatic increase in elicitation and quality of virus-neutralizing serum antibodies.
Stabilization of RSV F in prefusion conformation further enhances immunogenicity and protection.
Reduction of syncytium formation decreases cytopathology and selective pressure against RSV F expression.
Recombinant paramyxoviruses maintain stable expression of RSV F in vitro and in vivo despite enhanced expression and packaging.
Recombinant viruses exhibit efficient replication suitable for vaccine manufacture and clinical evaluation.
Bivalent immune response against both paramyxovirus and RSV provided by recombinant viruses.
Documented Applications
Use of the recombinant paramyxoviruses in immunogenic compositions or vaccines against parainfluenza viruses and respiratory syncytial virus.
Vaccination of subjects, especially infants and children, at risk of RSV and/or parainfluenza virus infection.
Administration by intranasal or parenteral routes to elicit protective immune responses.
Use in prime-boost immunization protocols to enhance immunity against RSV and parainfluenza viruses.
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