Recombinant chimeric bovine/human parainfluenza virus 3 expressing rsv g and its use
Inventors
Collins, Peter L. • Buchholz, Ursula J. • Liang, Bo • Munir, Shirin
Assignees
US Department of Health and Human Services
Publication Number
US-11975063-B2
Publication Date
2024-05-07
Expiration Date
2038-05-29
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Abstract
Recombinant chimeric bovine/human parainfluenza virus 3 (rB/HPIV3) vectors expressing Respiratory Syncytial Virus (RSV) G protein or a recombinant RSV G protein, as well as methods of their use and manufacture, are provided. The rB/HPIV3 vector comprises a genome comprising a heterologous gene encoding the RSV G protein or the recombinant RSV G protein. Nucleic acid molecules comprising the sequence of the genome or antigenome of the disclosed rB/HPIV3 vectors are also provided. The disclosed rB/HPIV3 vectors can be used, for example, to induce an immune response to RSV and HPIV3 in a subject.
Core Innovation
The problem addressed by the invention arises from the endemic burden of RSV causing severe lower respiratory tract diseases in infants, the elderly, and immunocompromised individuals. Despite demand, an effective RSV vaccine remains elusive, compounded by challenges including immature immune systems in infants, immune-suppression by maternal antibodies, inefficient superficial respiratory immune protection, and vaccine-induced enhanced disease observed with inactivated or subunit RSV and HPIV3 vaccines. Additionally, prior rB/HPIV3 vectors expressing RSV F antigen showed disappointing immunogenicity to RSV, insufficient for vaccine use.
Claims Coverage
The patent claims cover multiple inventive features related to a recombinant chimeric bovine/human parainfluenza virus 3 (rB/HPIV3) vector expressing RSV G protein variants and methods related to their use, manufacture, and immunogenic compositions.
Recombinant chimeric rB/HPIV3 vector genome composition
The rB/HPIV3 vector comprises a genome with a 3′ leader region, BPIV3 N gene, a heterologous gene encoding RSV G or recombinant variants, BPIV3 P and M genes, HPIV3 F and HN genes, BPIV3 L gene, and a 5′ trailer region. The heterologous gene encodes either a RSV G protein with ectodomain, transmembrane domain, and cytoplasmic tail, codon optimized for human expression; or recombinant RSV G protein with RSV G ectodomain linked to BPIV3, HPIV3, or HPIV1 HN transmembrane and cytoplasmic tail domains. The HPIV3 HN gene encodes an HN protein having threonine and proline residues at positions 263 and 370, respectively (SEQ ID NO:7). The vector is infectious, attenuated, and self-replicating.
Use of specific RSV G ectodomains and protein sequences
The RSV G ectodomain encoded consists of or is at least 90% identical to one of the sequences set forth as SEQ ID NOs: 23, 48, 50, 52, or 54. The recombinant RSV G protein includes transmembrane and cytoplasmic domains from BPIV3 HN, HPIV3 HN, or HPIV1 HN with at least 90% identity to SEQ ID NOs: 29, 57, or 60, respectively. The RSV G protein can be wild-type from subtype A or B RSV, with sequences or at least 90% identical to SEQ ID NOs: 22, 47, 49, 51, or 53.
Inclusion of BPIV3 and HPIV3 protein sequences
The BPIV3 N, P, C, V, M, and L genes encode proteins with at least 90% identity to sequences set forth as SEQ ID NOs: 1, 2, 3, 4, 5, and 10, respectively, and HPIV3 F and HN genes encode proteins at least 90% identical to SEQ ID NOs: 6 and 7, respectively.
Codon optimization for human expression and antigenomic cDNA
The heterologous RSV G gene is codon-optimized for expression in human cells and can comprise antigenomic cDNA sequences set forth as SEQ ID NOs: 89, 90, 91, or 92.
Induction of immune responses and neutralization
The rB/HPIV3 vector induces immune responses to RSV G protein, HPIV3 F protein, and HPIV3 HN protein, and elicits neutralizing antibodies against RSV and HPIV3.
Methods of production and use
The patent claims cover nucleic acid molecules comprising the viral genome or antigenomic cDNA or RNA, vectors containing these nucleic acids, host cells comprising the nucleic acids or vectors, methods for producing the recombinant virus by transfecting permissive cells, and immunogenic compositions containing the rB/HPIV3. Methods of eliciting immune responses in subjects via administration of these compositions, preferably intranasally, including in humans less than one year old, to generate protective immunity are claimed.
The claims comprehensively cover the recombinant chimeric bovine/human parainfluenza virus 3 vector comprising specific genetic configurations encoding RSV G protein variants, their nucleic acid molecules, production methods, immunogenic compositions, and methods for immunization that elicit neutralizing immune responses against RSV and HPIV3.
Stated Advantages
The rB/HPIV3-rsV G vectors are infectious, attenuated, and self-replicating, suitable for inducing protective immune responses to RSV and HPIV3 in pediatric subjects.
Inclusion of RSV G protein, especially the wild-type form expressed from the vector, induces serum RSV-neutralizing antibody titers comparable to wild-type RSV infection but at a lower dose and attenuation, indicating improved immunogenicity over prior constructs expressing RSV F protein.
Swapping transmembrane and cytoplasmic tail domains from PIV HN proteins into RSV G enhances packaging into viral particles, increasing antigen exposure and immune response.
Codon-optimization of the RSV G gene for human expression enhances protein expression and increases neutralizing antibody titers and protective efficacy.
Neutralizing antibodies induced by RSV G prevent RSV infection in a human airway epithelial model that expresses the CX3CR1 receptor, suggesting a better in vivo neutralization effect than antibodies to RSV F alone.
Documented Applications
Induction of immune responses to RSV and HPIV3 in subjects, including infants (1 to 18 months old) and other humans, to prevent or protect against RSV and HPIV3 infections.
Development and manufacture of recombinant viral vaccines for respiratory diseases caused by RSV and HPIV3.
Use in immunogenic compositions administered via intranasal routes to subjects at risk, including administration to seronegative infants and pediatric populations.
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