Recombinant human/bovine parainfluenza virus 3 (B/HPIV3) expressing a chimeric RSV/BPIV3 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-10654898-B2
Publication Date
2020-05-19
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
The invention provides recombinant paramyxoviruses comprising a viral genome encoding a heterologous gene that encodes an antigen of a heterologous virus. Specifically, the recombinant paramyxovirus can be a recombinant parainfluenza virus (PIV) including a genome encoding a heterologous respiratory syncytial virus (RSV) fusion (F) protein ectodomain linked to the transmembrane domain (TM) and the cytoplasmic tail (CT) of the F protein from the paramyxovirus. The recombinant viruses can be included in immunogenic compositions for eliciting bivalent immune responses to both the paramyxovirus and the heterologous RSV F protein.
The problem addressed by the invention arises from the difficulty in developing an effective RSV vaccine despite RSV's significant impact as a pathogen causing severe respiratory infections, especially in infants, children, and the elderly. Previous attempts to express RSV F protein from paramyxovirus vectors, e.g., Sendai virus, resulted in poor incorporation of RSV F into virus particles unless the vector's F protein was deleted, which is incompatible with generating infectious, attenuated vaccine viruses. Additionally, high levels of syncytium formation mediated by the RSV F protein can interfere with vector replication and cause genetic instability, reducing immunogenicity.
To overcome these issues, the invention surprisingly demonstrates that swapping the TM and CT domains of the heterologous RSV F protein with those from the paramyxovirus F protein dramatically increases incorporation of RSV F ectodomain into the envelope of the recombinant paramyxovirus. This increase in packaging enhances the elicitation of neutralizing serum antibodies in both quantity and quality when the recombinant paramyxovirus is administered. Furthermore, stabilizing the RSV F protein in the prefusion conformation further increases the induction of strongly neutralizing antibodies and protection. The invention also provides methods to optimize expression, attenuation, and stability, including codon optimization and selection of insertion sites within the viral genome.
Claims Coverage
The patent includes eleven independent claims covering recombinant paramyxoviruses and related compositions with heterologous genes encoding RSV F ectodomain linked to paramyxovirus F protein domains, as well as nucleic acids and methods for eliciting immune responses.
Recombinant paramyxovirus encoding chimeric RSV F protein
A recombinant paramyxovirus comprising a viral genome encoding PIV N, P, M, F, HN, L proteins, and a heterologous gene encoding a recombinant RSV F ectodomain linked to TM and CT domains of the PIV F protein, wherein the RSV F ectodomain linked to TM and CT is encoded by nucleotide sequences SEQ ID NO: 11, 22, or 23; and the recombinant virus is a recombinant human/bovine parainfluenza virus 3 (B/HPIV3), HPIV3, or BPIV3.
Specific recombinant HPIV3, BPIV3, or B/HPIV3 encoding defined RSV F sequences
Recombinant HPIV3, BPIV3, or B/HPIV3 viruses comprising viral genomes with specific gene sets and encoding the RSV F ectodomain linked to TM and CT domains as encoded by SEQ ID NO: 11, 22, or 23, with the B/HPIV3 encoding HPIV3 F and HN proteins plus BPIV3 other proteins.
Genomic location of heterologous gene
The heterologous gene encoding recombinant RSV F ectodomain is inserted as the first or second gene downstream of the genomic promoter within the paramyxovirus genome.
Recombinant HPIV3 or BPIV3 and B/HPIV3 with defined gene order and RSV F gene insertion
Recombinant HPIV3, BPIV3, or B/HPIV3 with viral genomes arranged from genomic promoter through N, P, M, F, HN, and L genes, with heterologous RSV F gene inserted between promoter and N or between N and P genes, encoding RSV F ectodomain linked to TM and CT domains encoded by SEQ ID NOs: 11, 22, or 23.
Conservative mutations in HPIV3 HN for vector stability
Recombinant B/HPIV3 or HPIV3 including mutations I263T and T370P in the HPIV3 HN protein to confer phenotypic stability.
Immunogenic compositions and methods of eliciting immune responses
Immunogenic compositions comprising the claimed recombinant paramyxoviruses and pharmaceutically acceptable carriers, and methods of eliciting immune responses to RSV and parainfluenza virus by administering such compositions to subjects.
The claims cover recombinant paramyxoviruses (notably B/HPIV3, HPIV3, and BPIV3) encoding heterologous RSV F ectodomains linked to paramyxovirus F protein TM and CT domains, with defined gene insertion positions and sequences, together with related compositions, nucleic acids, and immunogenic methods. Essential inventive features include the chimeric RSV F linkage improving incorporation and immunogenicity, codon optimization, and vector mutations enhancing stability and attenuation.
Stated Advantages
The chimeric RSV F protein with PIV TM and CT domains has multi-fold increased incorporation into recombinant paramyxovirus particles, improving antigen presentation.
Dramatic increase in quantity and quality of virus-neutralizing serum antibodies elicited in subjects after administration of the recombinant virus.
Stabilization of RSV F protein in the prefusion conformation further improves immunogenicity and induction of strongly neutralizing antibodies.
Insertion of heterologous RSV F gene and optimizations (such as codon optimization and insertion site selection) enhance expression without compromising replication needed for vaccine manufacture.
The recombinant viruses exhibit genetic stability of RSV F expression with reduced cytopathic effects due to reduced syncytium formation, stabilizing vaccine constructs.
Documented Applications
Use of the recombinant paramyxoviruses in vaccine formulations to elicit bivalent immune responses against parainfluenza virus (PIV) and respiratory syncytial virus (RSV).
Inducing protective immunity in subjects against RSV and PIV infections by administration of recombinant paramyxovirus vaccines.
Use in prime-boost vaccination strategies for controlling pediatric respiratory infections caused by RSV and parainfluenza viruses.
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