Generating expression vectors comprising myxoviral gene sequences for use as tool in developing vaccines to prevent infection and respiratory sysytem disorders; genetic vaccines and viral diseases
Inventors
SKIADOPOULOS, Mario H. • Murphy, Brian R. • Collins, Peter L.
Assignees
US Department of Health and Human Services
Publication Number
US-7820181-B2
Publication Date
2010-10-26
Expiration Date
2023-09-18
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Abstract
Recombinant human parainfluenza virus type 2 (HPIV2) viruses and related immunogenic compositions and methods are provided. The recombinant HPIV2 viruses, including HPIV2 chimeric and chimeric vector viruses, provided according to the invention are infectious and attenuated in permissive mammalian subjects, including humans, and are useful in immunogenic compositions for eliciting an immune responses against one or more PIVs, against one or more non-PIV pathogens, or against a PIV and a non-PIV pathogen. Also provided are isolated polynucleotide molecules and vectors incorporating a recombinant HPIV2 genome or antigenome.
Core Innovation
The invention provides methods and compositions for recovering infectious, recombinant human parainfluenza virus type 2 (HPIV2) and for introducing defined, predetermined structural and phenotypic changes into an infectious HPIV2 candidate. The recombinant HPIV2 viruses include complete viruses and subviral particles which may lack one or more non-essential viral proteins or portions thereof. The coexpression system involves expression of polynucleotide molecules encoding HPIV2 genome or antigenome RNA along with PIV proteins necessary for viral replication.
The problem being solved is the lack of approved vaccines for HPIVs and the challenges in developing effective tools and methods to engineer safe and effective immunogenic compositions against multiple HPIV serotypes and other pathogens. Existing vaccines are limited, and there is an urgent need for additional methods to generate suitably attenuated, immunogenic, and genetically stable HPIV2 and chimeric vectors. The invention addresses the challenge of producing recombinant HPIV2 and modifying it with specific mutations, insertions, deletions, or combinations thereof, to yield immunogenic compositions to prevent infections and respiratory diseases.
Claims Coverage
The patent claims include independent claims directed to recombinant HPIV2 compositions and isolated polynucleotides incorporating specific attenuating mutations, along with associated vectors. There are six inventive features extracted from the claims.
Recombinant HPIV2 with specific L protein mutations
An isolated, infectious, self-replicating recombinant human parainfluenza virus type 2 comprising PIV N, P, and L proteins and a partial or complete polyhexameric HPIV2 genome or antigenome encoding one or more attenuating amino acid substitutions or deletions in the L polymerase at the amino acid corresponding to residue 11 of SEQ ID NO. 4 and/or residue 11 of SEQ ID NO. 10.
Additional recombinantly-introduced attenuating mutations in HPIV2
The recombinant HPIV2 genome or antigenome further incorporates one or more additional recombinantly-introduced attenuating mutations.
Attenuating mutations corresponding to heterologous viruses
The recombinant HPIV2 genome or antigenome incorporates additional recombinantly-introduced attenuating mutations at amino acid positions corresponding to attenuating mutations identified in heterologous, mutant nonsegmented negative stranded RNA viruses.
Use of HPIV3 JS cp45 mutations in HPIV2 genome
The polynucleotide molecule encoding the recombinant HPIV2 genome or antigenome incorporates one or more mutations from HPIV3 JS cp45.
Isolated polynucleotide encoding HPIV2 with specific attenuating L mutations
An isolated polynucleotide comprising a partial or complete polyhexameric recombinant HPIV2 genome or antigenome modified by attenuating mutations introduced at any nucleotide encoding the amino acid corresponding to residue 11 of SEQ ID NO. 4 and/or residue 11 of SEQ ID NO. 10 in the HPIV2 L polymerase.
Expression vector comprising HPIV2 genome with specific attenuating mutations
An expression vector comprising an operably linked transcriptional promoter, a polynucleotide sequence comprising a partial or complete polyhexameric recombinant HPIV2 genome or antigenome modified by attenuating mutations at any nucleotide encoding the amino acid corresponding to residue 11 of SEQ ID NO. 4 and/or residue 11 of SEQ ID NO. 10 in the HPIV2 L polymerase, and a transcriptional terminator.
The claims cover recombinant HPIV2 viruses and isolated polynucleotides encoding HPIV2 genomes or antigenomes incorporating specific attenuating mutations in the L polymerase, including mutations corresponding to heterologous viruses such as HPIV3 JS cp45, and expression vectors comprising such modified HPIV2 genomes.
Stated Advantages
Recombinant HPIV2 vectors are infectious, attenuated, and immunogenic in humans and other mammals, useful for eliciting immune responses against one or more PIVs or non-PIV pathogens.
The system allows for generation with high frequency and fidelity of HPIV2 with defined genome sequences and predictable characteristics.
HPIV2 vectors have superior biological characteristics compared to other negative strand RNA virus vectors, including restriction of infection to the respiratory tract avoiding immunosuppression and infection of secondary organs.
The recombinant HPIV2 vectors enable fine adjustment of attenuation and immunogenicity through introduction of defined mutations, including temperature-sensitive and host range attenuating mutations.
Insertion of supernumerary genes into HPIV2 genome can attenuate virus and enable expression of protective antigens of heterologous pathogens.
Use of HPIV2 as a vector avoids limitations of other vectors such as measles virus and rabies virus, which have issues including immunosuppression, neurotoxicity, and poor suitability for infant immunization.
The rule of six ensures efficient replication and genetic stability in HPIV2, facilitating production of defined virus sequences.
Documented Applications
Use of recombinant HPIV2 viruses and chimeras to develop immunogenic compositions and vaccines to elicit immune responses against one or more HPIV serotypes, non-PIV pathogens including measles virus, respiratory syncytial virus (RSV), and human metapneumovirus.
Development of live attenuated vaccines and vectors for pediatric populations targeting respiratory infections caused by HPIVs, RSV, HMPV, measles virus, and other viral pathogens.
Use of chimeric HPIV2 as a vector to express heterologous viral antigen genes or genome segments as supernumerary genes for vaccination against multiple pathogens.
Sequential and coordinate immunization protocols employing recombinant HPIV vectors to elicit multispecific immune responses in early infancy to a range of respiratory pathogens including HPIV1, HPIV2, HPIV3, RSV, HMPV, and measles virus.
Transient gene therapy of the respiratory tract by recombinant HPIV2 vectors expressing therapeutic gene products, such as cytokines or proteins for treatment of diseases.
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