Sin Nombre virus full-length M segment-based DNA vaccines
Inventors
Assignees
United States Department of the Army
Publication Number
US-11155834-B2
Publication Date
2021-10-26
Expiration Date
2031-01-31
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
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 contemplates 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 was cloned into a plasmid to form the first stable recombinant SNV full-length M gene capable of eliciting neutralizing antibodies. The gene can be engineered into a vaccine system and is useful to protect mammals against infection with Sin Nombre virus.
The background identifies a critical problem: Hantaviruses, including Sin Nombre virus (SNV), cause hantavirus pulmonary syndrome (HPS), a highly lethal disease with rapid onset and progression, and a significant mortality rate of 35-40%. There are currently no specific therapeutics or vaccines to treat or prevent HPS caused by SNV or related hantaviruses. The lack of pre-existing immunity and the absence of effective medical countermeasures render populations vulnerable. Moreover, prior attempts to create vaccines that elicit neutralizing antibodies against SNV have been unsuccessful. This patent addresses the urgent need for an effective vaccine that produces a protective immune response against SNV infection.
The summary details the development of the codon-optimized SNV full-length M gene ORF, expressed in a plasmid (pWRG/SN-M(opt)), which stabilizes the mRNA and avoids plasmid destabilization issues in E. coli. Four amino acids unique to the original cloned sequence were changed to consensus residues to improve immunogenicity. This optimized M gene vaccine elicits high titers of neutralizing antibodies against SNV in animal models, which was previously unachieved. The invention also allows combination with other hantavirus M gene vaccines, such as Andes virus, permitting multivalent vaccines against multiple HPS and HFRS-causing hantaviruses.
Claims Coverage
The claims cover methods of inducing immune responses against Sin Nombre virus by administering recombinant DNA constructs encoding SNV glycoproteins, optionally combined with constructs from other hantaviruses, highlighting key plasmid constructs and delivery methods.
Method for inducing SNV immune response using codon-optimized M gene DNA constructs
A method administering a DNA vaccine composition comprising a plasmid having SEQ ID NO:2 or SEQ ID NO:3 encoding SNV Gc and Gn glycoproteins under a promoter, resulting in expression of these glycoproteins and eliciting an immune response in mammals or birds.
Use of specific promoters for SNV M gene expression
Inclusion of promoters such as cytomegalovirus promoter, beta-actin promoter, or SV40 promoter operably linked to the SNV nucleic acid sequence to drive expression of SNV glycoproteins.
Administration of effective DNA amounts via various delivery methods
Delivering from about 5 micrograms to about 5 milligrams of the DNA vaccine via methods including needle inoculation, needle-free jet injection, electroporation, or gene gun to elicit neutralizing antibody titers of at least 100, preferably at least 10,000.
Combination vaccination with Andes virus DNA vaccine plasmid
Co-administration of a second plasmid encoding Andes virus Gc and Gn glycoproteins (e.g., SEQ ID NO:10) to elicit immune responses against both SNV and Andes virus.
Multivalent vaccination including HFRS-associated hantaviruses
Administration of a third plasmid encoding one or more Gn and Gc glycoproteins from hantaviruses such as Hantaan, Puumala, and Seoul viruses (plasmids SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13 or nucleic acid SEQ ID NO:14), resulting in expression of these proteins and immune response induction.
The claims define innovative DNA vaccine methods employing codon-optimized SNV M gene constructs, optional multivalent formulations including Andes virus and other hantaviruses, specific promoter usage, and varied administration routes to induce potent SNV neutralizing immune responses in mammals and birds.
Stated Advantages
The vaccine elicits high-titer neutralizing antibodies against Sin Nombre virus, a first for any SNV vaccine.
The synthetic codon optimization increases mRNA stability and plasmid stability, improving immunogenicity and neutralizing antibody production.
The vaccine can be combined with hantavirus DNA vaccines for Andes virus and other HPS/HFRS hantaviruses, enabling multivalent protection.
The DNA vaccine platform allows for safer production without handling live virus, enhancing biosafety.
Documented Applications
Use as a vaccine to protect mammals and birds against infection with Sin Nombre virus by eliciting high-titer neutralizing antibodies.
Combination vaccines providing immunity against multiple HPS-associated hantaviruses (e.g., Andes virus) and HFRS-associated hantaviruses (e.g., Puumala, Hantaan, Seoul viruses).
Production of polyclonal and monoclonal antibodies for use as therapeutic or prophylactic agents in treating or preventing Sin Nombre virus infection.
Diagnostic use of antibodies raised against the SNV M gene products for detection of Sin Nombre virus infection in samples.
Production of pseudotyped viruses displaying SNV glycoproteins for serologic assays and gene therapy delivery to endothelial cells targeted by hantavirus glycoproteins.
Interested in licensing this patent?