Sin Nombre virus full-length M segment-based DNA vaccines
Inventors
Assignees
United States Department of the Army
Publication Number
US-10443073-B2
Publication Date
2019-10-15
Expiration Date
2031-01-31
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Abstract
The invention contemplates a new synthetic, codon-optimized Sin Nombre virus (SNV) full-length M gene open reading frame (ORF) that encodes a unique consensus amino acid sequence. The SNV ORF was cloned into a plasmid to form the first stable recombinant SNV full-length M gene that elicits neutralizing antibodies. The gene can be engineered into a vaccine system, and is useful to protect mammals against infection with Sin Nombre virus.
Core Innovation
The invention provides a novel synthetic, codon-optimized Sin Nombre virus (SNV) full-length M gene open reading frame (ORF) that encodes a unique consensus amino acid sequence. This SNV ORF has been cloned into a plasmid to form the first stable recombinant SNV full-length M gene plasmid, denoted pWRG/SN-M(opt), which is capable of eliciting neutralizing antibodies. The SNV M gene DNA sequence was engineered to increase mRNA stability and plasmid stability in E. coli, and to correct four amino acids to consensus sequences based on alignments with five hantavirus M gene sequences from GeneBank. This synthetic M gene was inserted into a plasmid-based DNA vaccine system and is believed to be the first vaccine of any kind to elicit convincing levels of neutralizing antibodies against Sin Nombre virus in animals.
The problem being solved is the need for effective medical countermeasures against hantavirus pulmonary syndrome (HPS) caused by Sin Nombre virus, which has a high case-fatality rate of approximately 35-40% and no existing vaccine or specific therapeutic treatments. Prior attempts to produce vaccines eliciting neutralizing antibodies against SNV have been unsuccessful, and existing vaccines for related hantaviruses do not reliably cross-neutralize SNV. There is an alarming lack of vaccines for HPS-associated hantaviruses like SNV, despite the significant health and biodefense threat posed by these viruses.
The invention thus addresses the need for a stable, codon-optimized full-length SNV M gene DNA vaccine that can induce high titer neutralizing antibodies and provide protective immunity against SNV infection. The design uses codon optimization and sequence consensus correction to produce an improved DNA vaccine capable of eliciting protective immune responses with fewer vaccinations than prior constructs. Additionally, the SNV DNA vaccine can be combined with other hantavirus M gene vaccines, such as Andes virus M gene vaccines, to formulate bivalent or multivalent vaccines protecting against multiple hantaviruses including those causing HPS and hemorrhagic fever with renal syndrome (HFRS).
Claims Coverage
The independent claims cover the composition and method of inducing neutralizing antibodies against Sin Nombre virus using a recombinant plasmid DNA vaccine.
Recombinant plasmid DNA expressing codon-optimized SNV M gene
The composition includes a recombinant plasmid DNA capable of expressing Sin Nombre virus Gc and Gn glycoproteins, where the plasmid DNA contains a modified codon-optimized full-length M gene selected from SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3 operably linked to a promoter resulting in expression of the glycoproteins.
Pharmacologically acceptable carrier in immunizing composition
The immunizing composition further includes a pharmacologically acceptable carrier such as PBS, water, saline, TRIS-EDTA, or mixtures thereof, facilitating administration of the DNA vaccine.
Immunizing amount of SNV plasmid DNA
The amount of SNV plasmid DNA in the composition is specified to be between about 5 micrograms and about 5 milligrams.
Combination with Andes virus M gene vaccine
The composition can further include an Andes M gene sequence operably linked to a promoter to express Andes hantavirus glycoproteins, allowing bivalent vaccination against SNV and Andes virus.
Combination with Puumala, Hantaan, and Seoul virus M gene vaccines
The composition can further comprise Puumala, Hantaan, and/or Seoul virus M gene sequences, each operably linked to promoters enabling expression, for broader hantavirus protection.
The claims encompass DNA vaccine compositions comprising codon-optimized SNV M gene sequences alone or in combination with other hantavirus M genes, delivered with a pharmacologically acceptable carrier, to elicit high titer neutralizing antibody responses against SNV and other hantaviruses.
Stated Advantages
The vaccine elicits high titer neutralizing antibodies against Sin Nombre virus, which was previously unachieved.
The codon optimization and consensus amino acid correction of the SNV M gene increases immunogenicity and reduces the number of vaccinations needed.
The DNA vaccine approach is inherently safe, avoids handling live Sin Nombre virus, and can be delivered by multiple methods including gene gun and electroporation.
The vaccine can be combined with other hantavirus DNA vaccines to produce multivalent vaccines protecting against multiple hantaviruses causing HPS and HFRS.
Documented Applications
Use of the codon-optimized SNV full-length M gene DNA vaccine to immunize mammals and birds against Sin Nombre virus infection.
Combination vaccines comprising SNV and other hantavirus M gene DNA vaccines (e.g., Andes virus, Puumala virus, Hantaan virus, Seoul virus) for bivalent or pan-hantavirus immunization.
Production of polyclonal and monoclonal antibodies in vaccinated animals for prophylactic or therapeutic treatment of SNV infection.
Development of diagnostic antibodies and immunoassays for detecting Sin Nombre virus infection using antibodies raised by the DNA vaccine.
Production of pseudotyped viruses incorporating the Sin Nombre virus glycoproteins by transfecting cells with the codon-optimized SNV M gene plasmid for serologic assays or targeted gene therapy delivery to endothelial cells.
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