The invention provides a recombinant biologically contained influenza virus that is a PB2 knockout virus, e.g., one that is useful to generate a multivalent vaccine, and methods of making and using that virus.

The disclosure provides for an isolated recombinant influenza virus having at least one of: a PB2 viral segment encoding PB2 with residue at position 540 that is not asparagine or a residue at position 712 that is not glutamic acid, a PA viral segment encoding PA with a residue at position 180 that is not glutamine or a residue at position 200 that is not threonine, or a PB1 viral segment encoding PB1 with a residue at position 149 that is not valine, a residue at position 634 that is not glutamic acid or a residue at position 635 that is not aspartic acid, or any combination thereof, and methods of making and using the virus.

Disclosed are delivery and expression systems of multiple antiviral therapeutic molecules. The therapeutic molecules include a novel class of dual-functional peptide and defective interfering genes of a virus. Also disclosed are compositions comprising the therapeutic molecules that are useful for the treatment and prevention of viral infections. Also disclosed herein are the method of making and using a vector that expresses the therapeutic molecules. Therapeutic molecules include cellular components such as RNA, DNA, peptide, proteins or combination thereof.

The invention provides a composition useful to prepare influenza vaccine viruses, e.g., in the absence of helper virus, which includes internal viral segments from an influenza virus vaccine strain or isolate, e.g., one that is safe in humans, for instance, one that does not result in significant disease, and encodes a heterologous antigen.

Disclosed are compositions and methods comprising one or more recombinant influenza viruses. Recombinant influenza viruses with mutated polymerases and/or rearranged genomes are disclosed. Constructs comprising different influenza nucleic acid sequences are also provided. Methods of inducing protecting immunity with the recombinant influenza viruses are disclosed. Also disclosed are methods of plasmid-free production of influenza virus comprising amplicons comprising one or more of influenza genes.

Disclosed herein are compositions and methods related to mutant viruses, and in particular, mutant influenza viruses. The mutant viruses disclosed herein include a mutant M2 sequence, and are useful in immunogenic compositions, e.g., as vaccines. The mutant viruses disclosed herein including a mutant M2 sequence are also useful to deliver antigens to a subject, e.g., to induce an immune response to the antigen. Also disclosed herein are methods, compositions and cells for propagating the viral mutants, and methods, devices and compositions related to vaccination.

The development of a computationally optimized influenza HA protein that elicits broadly reactive immune response to all H5N1 influenza virus isolates is described. The optimized HA protein was developed through a series of HA protein alignments, and subsequent generation of consensus sequences, for clade 2 H5N1 influenza virus isolates. The final consensus HA amino acid sequence was reverse translated and optimized for expression in mammalian cells. Influenza virus-like particles containing the optimized HA protein are an effective vaccine against H5N1 influenza virus infection in animals.

Embodiments relate to a method comprising (a) expressing a vector comprising a PSGL-1 (P-selectin glycoprotein ligand-1) or a mutant thereof in a VPC (virus producing cell); and blocking a virus infection by inactivating an infectivity of a released virions from the VPC; or (b) expressing a glycoprotein or a mutant thereof in the VPC; blocking the virus infection by preventing binding of the released virions to a target cell; inactivating infectivity of the released virions; and targeting a viral infection. Other embodiments relate to (1) a broad-spectrum anti-viral product comprising: a vector expressing a glycoprotein or a mutant thereof in a VPC; and blocking a virus infection by inactivating infectivity of a released virion from the VPC; and (2) a vaccine comprising a viral particle is configured to a live attenuated or an inactivated or a non-infectious, wherein the viral particle are produced in a VPC.

The invention provides a recombinant biologically contained influenza virus that is a PB2 knockout virus, e.g., one that is useful to generate a multivalent vaccine, and methods of making and using that virus.

The disclosure provides for an isolated recombinant influenza virus having at least one of: a PB2 viral segment encoding PB2 with residue at position 540 that is not asparagine or a residue at position 712 that is not glutamic acid, a PA viral segment encoding PA with a residue at position 180 that is not glutamine or a residue at position 200 that is not threonine, or a PB1 viral segment encoding PB1 with a residue at position 149 that is not valine, a residue at position 684 that is not glutamic acid or a residue at position 685 that is not aspartic acid, or any combination thereof, and methods of making and using the virus.