Disclosed herein is a method for producing human antibodies against a pathogen comprising injecting a non-human animal with a pathogen-derived DNA vaccine in at least two locations of the animal; injecting the animal with an adjuvant in a location of the animal different from the location of the DNA vaccine location; collecting plasma from the animal after the injections; and purifying polyclonal antibody from the plasma.

Disclosed herein is a method for producing human antibodies against a pathogen comprising injecting a non-human animal with a pathogen-derived DNA vaccine in at least two locations of the animal; injecting the animal with an adjuvant in a location of the animal different from the location of the DNA vaccine location; collecting plasma from the animal after the injections; and purifying polyclonal antibody from the plasma.

The present invention describes specific engineering of the hantavirus spike proteins with modifications to stabilize (Gn/Gc).sub.n heterodimer contacts and/or Gc homodimer contacts and/or Gn/Gn oligomer contacts on the spike with the purpose of using them as immunogens in next-generation vaccine design. Said spike proteins have been covalently stabilized by at least one disulphide inter-chain bond between Gn/Gc heterodimers and/or between Gc homodimers and/or between Gn homo-oligomers as they are presented at the surface of infectious virions. It also involves spike stabilization by introduction of cavity-filling amino acids with a bulky side chain at the above-mentioned contacts. Said spike proteins can be soluble Gn/Gc ectodomains in solution and/or incorporated as (Gn/Gc).sub.n hetero-oligomers onto virus-like particles (VLPs) and/or used for pseudotyping virus vectors and/or form part of a stabilized recombinant virus, wherein said spike proteins can be used to select ligands and/or can be used for preventing or treating infections by one or more hantaviruses.

The present invention provides a viral vector or bacterial vector, said vector comprising a nucleic acid sequence encoding a Hantavirus nucleoprotein or antigenic fragment thereof; wherein said vector is capable of inducing a protective immune response in a subject. The present invention also provides compositions and uses of the vector in methods of medical treatment.

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.

A nucleic acid vaccine composition comprising one or more of a plasmid-based nucleic acid vaccine and immunotherapy, as well as a lipid formulation, is provided. In addition, the present invention provides a method of enhancing the potency of plasmid-based DNA vaccines and immunotherapies, by formulating a vaccine and/or immunotherapy in a lipid formulation, which is stable when refrigerated or stored frozen, is then delivered to a vaccinee by either needle/syringe, jet injection, or microneedles. The lipid formulation of the present invention comprises one or more lipid excipients selected from 1,2-Distearoyl-sn-glycero-3-phosphocholine, Cholest-5-en-3β-ol, 1,2-Dimyristoyl-rac-glycero-3-methylpolyoxyethlene, and or more symmetric ionizable cationic lipids. The present invention increases vaccine potency dramatically. It was unexpectedly discovered that the level of immunogen, or immune response molecules, produced in vivo is increased (versus administering merely the vaccine or immunotherapy) and, in the case of a vaccine immunogen, the immune response is enhanced.

Hantavirus spike proteins with modifications to stabilize (Gn/Gc)n heterodimer contacts and/or Gc homodimer contacts and/or Gn/Gn oligomer contacts on the spike to enable their use as immunogens in next-generation vaccine design. The spike proteins have been covalently stabilized by at least one disulphide inter-chain bond between Gn/Gc heterodimers and/or between Gc homodimers and/or between Gn homo-oligomers as they are presented at the surface of infectious virions. Also, spike stabilization by introduction of cavity-filling amino acids with a bulky side chain at the above-mentioned contacts. The spike proteins can be soluble Gn/Gc ectodomains in solution and/or incorporated as (Gn/Gc)n hetero-oligomers onto virus-like particles (VLPs) and/or used for pseudotyping virus vectors and/or form part of a stabilized recombinant virus. The spike proteins can be used to select ligands and/or can be used for preventing or treating infections by one or more hantaviruses.

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.

A nucleic acid vaccine composition comprising one or more of a plasmid-based nucleic acid vaccine and immunotherapy, as well as a lipid formulation, is provided. In addition, the present invention provides a method of enhancing the potency of plasmid-based DNA vaccines and immunotherapies, by formulating a vaccine and/or immunotherapy in a lipid formulation, which is stable when refrigerated or stored frozen, is then delivered to a vaccinee by either needle/syringe, jet injection, or microneedles. The lipid formulation of the present invention comprises one or more lipid excipients selected from 1,2-Distearoyl-sn-glycero-3-phosphocholine, Cholest-5-en-3-ol, 1,2-Dimyristoyl-rac-glycero-3-methylpolyoxyethlene, and or more symmetric ionizable cationic lipids. The present invention increases vaccine potency dramatically. It was unexpectedly discovered that the level of immunogen, or immune response molecules, produced in vivo is increased (versus administering merely the vaccine or immunotherapy) and, in the case of a vaccine immunogen, the immune response is enhanced.

A synthetic, codon-optimized Hantaan virus (HTNV) full-length M gene open reading frame that consists of a unique nucleotide sequence encoding HTNV proteins. This synthetic gene was cloned into a plasmid to form the first optimized HTNV full-length M gene that elicits neutralizing antibodies in animals when delivered in combination with a similarly optimized Puumala virus (PUUV) DNA vaccine. The invention obviates the need for an extraneous gene sequence that was previously required for expression of the non-optimized HTNV gene. The synthetic gene is engineered into a molecular vaccine system to prevent hemorrhagic fever with renal syndrome (HFRS) caused by infection with HTNV, SEOV, or DOBV. Alternatively, it can be combined with the optimized PUUV DNA vaccine to protect against HFRS caused by any hantavirus.