Swine influenza vaccine compositions and methods thereof

Abstract

The present disclosure provides novel sequences, vectors and pharmaceutical compositions for the treatment of IAV in swine. Also provided are methods of using the novel sequences, vectors, and pharmaceutical compositions comprising inducing an immune response in an animal and preventing IAV infection in an animal.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application Serial No. 63 / 433,242, filed on December 16, 2022, the entire disclosure of which is incorporated herein by reference.

[0003] Reference to electronically submitted sequence listings

[0004] The computer-readable nucleotide / amino acid sequence listing, which is incorporated herein by reference in its entirety, is submitted with this application and is confirmed as follows: XML file named "85007_399822.xml" created on December 15, 2023 (36kb). Background Art

[0005] Influenza A viruses are diverse and deadly pathogens of pigs. Currently circulating endemic subtypes of influenza A viruses in pigs include H1N1, H1N2, and H3N2. The evolutionary history of swine influenza viruses (IAV-S) reflects multiple introductions of influenza viruses into pigs from other species, such as avian species and humans. For example, the H1N1 subtype known as classical H1N1 virus (cH1N1) was first isolated from pigs in 1930 and subsequently dominated North America until the emergence and widespread spread of recombinant H3N2 in 1998.

[0006] Swine influenza is one of the leading causes of acute respiratory illness outbreaks in pigs. Vaccination is the most common strategy for controlling swine influenza, but conventional approaches to developing effective vaccines have had limited success due to the large antigenic diversity of strains circulating in different parts of the world. Currently, only one RNA-based vaccine is licensed by the USDA for use in pigs, which demonstrates a similar level of protection as the licensed whole-inactivated virus (WIV). Notably, human influenza vaccines are updated annually to ensure the closest match to circulating strains, but swine influenza vaccines are outdated and contain older strains that are no longer prevalent in pigs. Therefore, new pharmaceutical compositions and methods for combating porcine IAV are urgently needed. Summary of the Invention

[0007] Thus, the present disclosure provides novel sequences, vectors, and pharmaceutical compositions for treating porcine IAV. Methods of using the novel sequences, vectors, and pharmaceutical compositions are also provided, including methods for inducing an immune response in an animal and methods for preventing IAV infection in an animal.

[0008] Compared to the prior art, the compositions and methods disclosed herein provide multiple benefits. First, certain polypeptide and nucleotide sequences provided herein represent consensus sequences that provide protection against different influenza viruses in pigs. Second, certain polypeptide and nucleotide sequences provided herein represent chimeric sequences that provide protection against different influenza viruses in pigs. Third, the vectors provided herein are capable of inducing high levels of antibodies and T cell responses to IAV in pigs. Additionally, the vectors described herein are capable of providing protection against homologous challenge, demonstrating their versatility in controlling IAV in pigs.

[0009] Through the following detailed description, other purposes, features and advantages of the present disclosure will become apparent. However, it should be understood that the detailed description and specific examples, although setting forth specific embodiments of the present invention, are given by way of illustration only, as various changes and modifications within the spirit and scope of the present invention will be apparent to those skilled in the art based on this detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The detailed description makes particular reference to the accompanying drawings, in which: Figures 1A to 1F Display of recombinant ORFV Δ121 Production of conH1. Figure 1A Shown is a phylogenetic tree representing the position of conH1 constructs (red dots) relative to the HA sequence used for their design. Figure 1B Shown are box plots illustrating the genetic distance between circulating strains used to design the consensus HA sequence (red) and how this distance can be reduced based on the consensus HA (yellow) for the H1 subtype. Figure 1C pUC57-ORFV is shown Δ121 Schematic diagram of the conH1-loxP-EGFP transfer plasmid and ORFV genome, which depicts the methods used to generate recombinant ORFV. Δ121 The ORFV121 insertion site and flanking regions (left flank - LF and right flank - RF) of conH1 were constructed. After homologous recombination, the resulting viral genome contained the common H1 and GFP reporter genes inserted into the ORFV121 locus. After viral screening, GFP was removed from the 121 locus using Cre recombinase to obtain the final recombinant ORFV121. Δ121 conH1, which exclusively delineates the consensus H1 sequence into the ORFV121 locus. Figure 1D The results show that the ORF121 gene sequence (401 bp) was amplified by PCR in wild-type virus and its expression in recombinant ORFV Δ121 Agarose gel of deletions in conH1. Figure 1E Is displayed from ORFV Δ121Agarose gel showing PCR amplification of the conH1 gene (1774 bp) from the conH1 virus and its absence in the wild-type viral genome. The negative control in the image is from a corresponding PCR reaction using MiliQ water instead of DNA. Figure 1F The results show that the recombinant ORFV Δ121 Western blot analysis of conH1 expression (approximately 70 kDa) was performed on OFTu cells infected at an MOI of 5 and harvested at 12, 24, 48, and 72 hpi in vitro. Cell lysates from uninfected OFTu cells were used as negative controls. Blots were developed with a Flag-tag epitope-specific mAb (created using Biorender.com). Figure 1C and assembly diagram).

[0011] Figures 2A to 2B Recombinant ORFV is shown Δ121 Replication kinetics of conH1. Figure 2A Recombinant ORFV in primary OFTu cells is shown Δ121 Multi-step and single-step growth curves of conH1 compared with wild-type virus. Figure 2B Recombinant ORFV in primary STu cells is shown Δ121 Multi-step and single-step growth curves of conH1. Virus titers were determined by the method of Spearman and Karber and expressed as the median tissue culture infectious dose (TCID 50 ) / ml (Figure created using Biorender.com).

[0012] Figures 3A to 3B ORFV Δ121 Localization of heterologous conH1 protein expressed by conH1. Figure 3A Immunofluorescence assays in permeabilized and non-permeabilized OFTu cells are shown. 48 h after infection at an MOI of 1, the recombinant virus expressed conH1 (red fluorescence) in the intracellular compartment (left) and on the cell membrane (right). Blue fluorescence indicates nuclear staining with DAPI. Figure 3B The expression of ORFV was assessed by flow cytometry. Δ121 Heterologous proteins expressed by conH1. Δ121 OFTu cells were infected with conH1 or wild-type OV IA82 as a negative control. Infected cells were harvested 48 hours post-infection, fixed, permeabilized or not, and stained with appropriate antibodies for flow cytometry analysis. The gate indicated by the rectangle aggregates positively stained cells (assembly map created using Biorender.com).

[0013] Figures 4A to 4C Different porcine antisera and ORFV are shownΔ121 Cross-reactivity between conH1. Figure 4A Figure 3 shows the cross-reactivity between sera from pigs infected with three different H1N1 IAV-S strains and conH1 expressed on the cell surface of OFTu permeabilized cells as assessed by IFA. Positive cross-reactivity is indicated by green fluorescence, indicating that conH1 is expressed by the recombinant ORFV. Δ121 conH1 virus-expressed conH1 is recognized by host sera from infected animals. Figure 4B Table 1 is an information table on the IAV-S isolates used for cross-reactive serological assays, including pairwise identities. Figure 4C A phylogenetic tree based on the nucleotide sequences of HA from these same isolates is shown to assess the genetic distance from conH1.

[0014] Figures 5A to 5E showed a humoral response to immunization. Figure 5A ) and CA / 09( Figure 5B In ) challenged pigs, the viruses used for challenge (OH / 07 and CA / 09) were used as coating antigens and the expression of ORFV was assessed by whole virus ELISA on DPC0 (before challenge). Δ121 conH1 induced IAV-S IgG response. Figure 5C ) and CA / 09( Figure 5D ) In the challenge group, for a group of 10 different viruses, whole virus ELISA was used to explore the ORFV Δ121 Breadth of conH1-induced IAV-S IgG immune response. Figure 5E A phylogenetic tree showing the distance between the HA of the viruses used for ELISA assays and the designed consensus sequence (common_H1) is shown. Each sample was tested in duplicate wells. Asterisks indicate statistical significance between the two animal groups. P values: P < 0.05, P < 0.01, P < 0.001, P < 0.0001.

[0015] Figures 6A to 6H Figure 2 shows the evaluation of T cell responses in PBMCs by FACS. The percentage of IFNγ+ T helper / memory cells ( Figure 6A ), IFNγ +CTL cell percentage ( Figure 6D ), IL-17A + CTL cell percentage ( Figure 6C ) and the percentage of IFNγ + CTL cells ( Figure 6DThe same T cell populations were also analyzed in PBMC isolated from pigs at DPC6 after CA / 09 recall stimulation at 0.1 MOI ( Figures 6E to 6H ). Asterisks indicate statistical significance between two animal groups. P values: P < 0.05, P < 0.01, P < 0.001, P < 0.0001.

[0016] 7A to 7D The T cell immune response to immunization is shown. The percentage of IL-17A + T lymphocytes in BAL MNC cells isolated from pigs at DPC6 after recall stimulation with OH / 07 at 0.1 MOI was analyzed. Figure 7A ) and the percentage of IFNγ + T lymphocytes ( Figure 7B The same T cell populations were also analyzed in BAL MNC isolated from pigs at DPC6 after recall stimulation with CA / 09 at an MOI of 0.1 ( Figure 7C and Figure 7D ). Asterisks indicate statistical significance between two animal groups. P values: P < 0.05, P < 0.01, P < 0.001, P < 0.0001.

[0017] Figures 8A to 8D The T cell immune response to immunization is shown. The percentage of IL-17A + T lymphocytes in TBLN MNC cells isolated from pigs at DPC6 after recall stimulation with OH / 07 at 0.1 MOI was analyzed. Figure 8A ) and the percentage of IFNγ + T lymphocytes ( Figure 8B The same T cell populations were also analyzed in TBLN MNC isolated from pigs at DPC6 after recall stimulation with CA / 09 at 0.1 MOI ( Figure 8C and Figure 8D ). Asterisks indicate statistical significance between two animal groups. P values: P < 0.05, P < 0.01, P < 0.001, P < 0.0001.

[0018] Figures 9A to 9B Viral loads in nasal swabs assessed by infectious virus titration and qRT-PCR are shown. Figure 9AVirus shedding in OH / 07 challenged pigs was demonstrated by infectious titers and genome copy number. Figure 9B Virus shedding in CA / 09-challenged pigs is shown by infectious titer and genome copy number. Asterisks indicate statistical significance between the two animal groups. P values: P < 0.05, P < 0.01, P < 0.001, P < 0.0001.

[0019] 10A to 10D Shown are the results of sham / challenge and ORFV evaluation at DPC6. Δ121 Lung lesion scores, viral infectivity titers, and viral loads in BAL and lung lysates of conH1 / challenged pigs. Figure 10A Scores when DPC6 was assessed are shown. Figure 10B Virus infection titer TCID 50 IAV-S viral RNA shedding in porcine BAL determined by RT-qPCR genome copy number / ml. Figure 10C Sham immunization and ORFV challenged with OH / 07 are shown Δ121 Virus infectious titers and genome copy numbers in the lungs of conH1-immunized pigs at DPC6. Figure 10D Shown are mock immunization and ORFV challenged with CA / 09 Δ121 Virus infection titers and genome copy numbers in the lungs of conH1-immunized pigs at DPC 6. Asterisks indicate statistical significance between the two animal groups. P values: P < 0.05, P < 0.01, P < 0.001, P < 0.0001.

[0020] 11A to 11D An overview of the chimeric vaccine strategy is shown. Figure 11A Shown are sequential immunizations with chimeric-HA constructs consisting of a foreign head and a shared identical stalk domain belonging to the H1 subtype. Figure 11B Shown is a phylogenetic tree representing two groups within the H1 subtype of influenza A. The arrow indicates the subtype used to design the chimeric HA head. Figure 11C Indicates the construction for generating ORFV by homologous recombination Δ121 Recombinant plasmid of cH6 / 1 recombinant virus. Figure 11D Indicates the construction for generating ORFV by homologous recombination Δ121 Recombinant plasmid of cH8 / 1 recombinant virus.

[0021] 12A to 12E ORFV is shown Δ121 cH6 / 1 and ORFV Δ121 Heterologous protein expression of cH8 / 1 recombinant virus. Recombinant ORFV was expressed in permeabilized and non-permeabilized cells (48 h post-infection) infected at an MOI of 1 as assessed by IFA. Δ121 cH6 / 1( Figure 12A ) and ORFV Δ121 cH8 / 1 ( Figure 12B ) of chimeric HA expression (red fluorescence). Figure 12C ORFV detected by Western blot is shown Δ121 The levels of cH6 / 1 (approximately 70 kDa) were increased in cH6 / 1-infected sheep cells. Figure 12D ORFV detected by Western blot is shown Δ121 Levels of cH8 / 1 are increased in cH8 / 1 infected sheep cells. For Western blotting, mock infected OFTu cells were used as a negative control, and β-actin was used as a loading control. Figure 12E and Figure 12F Shows the Figure 12C and Figure 12D The flow cytometry data were consistent with those of the assay, which showed that 72 hours after infection, cH6 / 1 ( Figure 12E ) was more expressed than cH8 / 1( Figure 12F ) was slightly higher. OV-IA82-infected OFTu cells were used as a negative control for the gating strategy, where gated cells indicated the positive cell population.

[0022] 13A to 13C ORFV is shown Δ121 cH6 / 1 and ORFV Δ121 Characterization of cH8 / 1-IAV-S. Figure 13A An agarose gel showing amplification of the H1 stalk fragment confirming its presence and the absence of the ORF121 gene indicates recombinant ORFV Δ121 cH6 / 1 and ORFV Δ121 The cH8 / 1 virus was purified. Figure 13B and Figure 13C Comparison of recombinant ORFV in OFTu cells and STu cells is shown. Δ121 cH6 / 1 and ORFV Δ121 Growth curves of the cH8 / 1 virus and the parental OV-IA82 virus at different time points.

[0023] FIG. 14A to FIG. 14BThe kinetics of the antibody response to immunization are shown. Whole virus ELISA was used to detect the antibody response to the challenge virus Oh07 ( Figure 14A ) and Ca09( Figure 14B ) evaluated the use of ORFV in Oh07 and Ca09 challenged pigs. Δ121 cH6 / 1 priming and ORFV Δ121 cH8 / 1-IAV-S boost induced an increase in IgG responses. P value: P < 0.05, P < 0.01, P < 0.001, P < 0.0001.

[0024] FIG. 15A to FIG. 15B Shown are the humoral responses to immunization on day 35 assessed using whole virus ELISA against a panel of 10 different viruses. Oh07( Figure 15A ) and Ca09( Figure 15B ) IgG antibody levels in animals challenged with ORFVΔ121cH6 / 1 as a primary immunization and then boosted with ORFVΔ121cH8 / 1-IAV-S. P value: P < 0.05, P < 0.01, P < 0.001, P < 0.0001.

[0025] 16A to 16B Shown are mucosal antibody responses in BAL on day 42 to immunization assessed using whole virus ELISA against a panel of 10 different viruses. Oh07( Figure 16A ) and Ca09( Figure 16B ) ORFV in challenged animals Δ121 cH6 / 1 priming and ORFV Δ121 IgA antibody levels induced by cH8 / 1 boost. P value: P < 0.05, P < 0.01, P < 0.001.

[0026] 17A to 17B Figure 3 shows IAV-S shedding through nasal secretions. The Oh07 challenged pigs were identified by infectious titer (top) and genome copy number (bottom). Figure 17A ) and Ca09( Figure 17B ) Virus shedding in challenged pigs. P value: P < 0.05, P< 0.01.

[0027] 18A to 18B Lung lesion scores and viral loads in target tissues are shown. Figure 18A Shown are lung lesions and localized presence of genomic RNA in pigs challenged with Oh07. Figure 18B Lung lesion scores and viral loads were assessed in Ca09-challenged animals. P < 0.05, P < 0.01.

[0028] 19A to 19D Figure 3 shows the results from piglets challenged with Oh07 virus strain vaccinated with OV-cH6 / 1-ORFV-cH8 / 1 ( Figure 19A ) and after sham immunization ( Figure 19B ), and piglets challenged with Ca09 virus strain were vaccinated with OV-cH6 / 1-ORFV-cH8 / 1 ( Figure 19C ) and sham immunity ( Figure 19D Images of macroscopic lesions found in the lungs after immunization with sham-immunized animals. Signs of tissue consolidation can be identified by dark, speckled areas, which are more pronounced in sham-immunized animals. DETAILED DESCRIPTION

[0029] Various embodiments of the present invention are described herein below. Various embodiments utilize one or more of the following sequences shown in Table 1. Optionally, the 'ATG' start codon and the 'TAA' stop codon described in the nucleic acid sequences in the table below may be omitted.

[0030] Table 1. Sequences

[0031] In exemplary aspects, amino acid sequences comprising consensus H1 hemagglutinin polypeptides are provided. An "amino acid sequence" refers to a sequence of amino acids in a protein and includes sequences in which one or more amino acid side groups have been chemically modified, as well as sequences in which one or more amino acids have been substituted, inserted, or deleted relative to a known sequence without thereby eliminating the desired property. An amino acid sequence may also be referred to as a peptide, oligopeptide, or protein.

[0032] A consensus H1 hemagglutinin polypeptide refers to the full-length coding sequence of HA from the H1 subtype. Such polypeptides can be generated in silico by analyzing and aligning a large panel of isolated IAV sequences of the H1 subtype obtained from clinically diagnosed cases. Similarly, a consensus H3 hemagglutinin polypeptide refers to the full-length coding sequence of HA from the H3 subtype. Such polypeptides can be generated in silico by analyzing and aligning a large panel of isolated IAV sequences of the H3 subtype obtained from clinically diagnosed cases.

[0033] In embodiments, the consensus H1 hemagglutinin polypeptide comprises SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 1.

[0034] In any embodiment relevant to the sequence listing described herein, the phrase "comprising" can be replaced by the phrase "consisting of, according to fully established principles. As used herein, "sequence identity" and "homology" with respect to polypeptide or amino acid sequences are defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a particular peptide or polypeptide sequence, after alignment of the sequences and introduction of gaps, to obtain maximum percentage sequence identity, if desired, and without considering any conservative substitutions as part of sequence identity. The same concept applies to nucleic acid or nucleotide sequences. Alignments for determining percent sequence identity can be achieved in various ways within the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or MEGALINE™ (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm required for achieving maximum alignment over the entire length of the compared sequence.

[0035] In exemplary aspects, nucleotide sequences comprising a nucleic acid sequence encoding a consensus H1 hemagglutinin polypeptide are provided. Nucleotide sequences encoding all or part of a polypeptide with or without a signal sequence are provided. Typically, the nucleotide sequence encoding the polypeptide of interest is inserted into an expression vector suitable for expression in a selected host cell.

[0036] In embodiments, the nucleic acid sequence comprises SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 2.

[0037] In an embodiment, the nucleic acid sequence is a native sequence modified to contain codons optimized for expression in prokaryotes.

[0038] In embodiments, the consensus H1 hemagglutinin polypeptide comprises SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 1. In embodiments, the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 1.

[0039] In exemplary aspects, a recombinant polynucleotide is provided that comprises a heterologous regulatory element operably linked to a nucleic acid sequence encoding a consensus H1 hemagglutinin polypeptide.

[0040] The aforementioned embodiments of the consensus H1 hemagglutinin polypeptide are applicable to the recombinant polynucleotides described herein. The aforementioned embodiments of the nucleic acid sequences are applicable to the recombinant polynucleotides described herein. In embodiments, the heterologous regulatory element is a promoter functional in prokaryotes.

[0041] The term "regulatory element" (also referred to as "regulatory region" or "regulatory sequence") refers to a nucleic acid sequence that directly or indirectly promotes and / or controls gene expression and / or protein expression. A regulatory element can be a promoter, enhancer, silencer, or a nucleic acid sequence encoding a microRNA (miRNA) or transcription factor. A regulatory element can increase or decrease gene expression and / or protein expression.

[0042] In some embodiments, regulatory elements bind to regulatory proteins (e.g., transcription factors) to control gene expression and / or protein expression. In some embodiments, regulatory elements encode transcription factors that control gene expression and / or protein expression. In some embodiments, regulatory elements encode miRNAs that bind to target mRNAs to control protein expression.

[0043] In some embodiments, the regulatory element is a controllable regulatory element. In some embodiments, the regulatory element is an uncontrollable regulatory element, such as a constitutive promoter. In some embodiments, the regulatory element is a positive regulatory element, such as a promoter. In some embodiments, the regulatory element is a negative regulatory element, such as a silencer. In some embodiments, the regulatory element provides transient, inducible (e.g., tetracycline responsive promoter, or hypoxia inducible promoter) and / or tissue-specific gene expression and / or protein expression.

[0044] In some embodiments, regulatory elements are operably linked to nucleic acids encoding polypeptides of the present disclosure (coding sequences). Regulatory elements need not be contiguous with coding sequences, as long as their function is to direct the expression of the encoded polypeptides. Thus, for example, there may be intermediate untranslated but transcribed sequences between a promoter sequence and a coding sequence, and the promoter sequence may still be considered "operably linked" to the coding sequence.

[0045] In some embodiments, the regulatory element is not operably linked to the nucleic acid encoding the polypeptide of the present disclosure. For example, the regulatory element can be a microRNA sequence or a transcription factor expressed from the same vector as the nucleic acid encoding the polypeptide or from a different vector.

[0046] In exemplary aspects, compositions are provided that include a consensus H1 hemagglutinin polypeptide and a carrier.

[0047] In embodiments, the consensus H1 hemagglutinin polypeptide comprises an amino acid sequence. In embodiments, the amino acid sequence comprises SEQ ID NO: 1. In embodiments, the amino acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 1. In embodiments, the amino acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 1. In embodiments, the amino acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 1. In embodiments, the amino acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 1. In embodiments, the amino acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 1. In embodiments, the amino acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 1. In embodiments, the amino acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 1. In embodiments, the amino acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 1.

[0048] In embodiments, the consensus H1 hemagglutinin polypeptide comprises a nucleic acid sequence encoding a consensus H1 hemagglutinin polypeptide. In embodiments, the nucleic acid sequence comprises SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 2. In embodiments, the nucleic acid sequence is a native sequence modified to include codons optimized for expression in prokaryotic cells.

[0049] The term "vector" is used to describe a polynucleotide that can be engineered to contain one or more cloned polynucleotides that can be amplified in a host cell. A vector can include one or more of the following elements: an origin of replication, one or more regulatory elements (e.g., a promoter or enhancer) that regulate expression of a polypeptide of interest, or one or more selectable marker genes (e.g., antibiotic resistance genes and genes useful for colorimetric assays, such as β-galactosidase). The term "expression vector" refers to a vector used to express a polypeptide of interest in a host cell.

[0050] "Host cell" refers to a cell that can be or has been a recipient of a vector or isolated polynucleotide. The host cell can be a prokaryotic cell or a eukaryotic cell. Exemplary eukaryotic cells include mammalian cells, such as primate or non-primate cells; fungal cells, such as yeast; plant cells; and insect cells. Non-limiting exemplary mammalian cells include, but are not limited to, NSO cells, PER.C6® cells (Crucell), 293 cells and CHO cells and their derivatives, such as 293-6E, DG-44, CHO-S and CHO-K cells. Host cells include the offspring of a single host cell, and due to natural, accidental or deliberate mutations, the offspring may not necessarily be exactly the same as the original parent cell (morphologically or in genomic DNA complementation). Host cells include cells transfected in vivo with polynucleotides encoding the amino acid sequences provided herein.

[0051] The vector can be a nucleic acid plasmid that can be delivered via non-viral methods (e.g., naked DNA / RNA, formulated DNA / RNA, or liposomes) or via viral methods. In some embodiments, the vector is a viral vector, such as a retroviral vector, a herpes virus vector, an adenoviral vector, an adeno-associated virus vector, or a pox virus vector. The vector can be a bacterial vector.

[0052] In an embodiment, the vector is a viral vector. In an embodiment, the viral vector is an ORFV vector. Capripox virus (ORFV) is a large DNA virus that can accommodate viral antigens and effectively deliver them to animals.

[0053] In an embodiment, the ORFV vector does not comprise ORFV121.

[0054] In an embodiment, the vector is a nucleic acid vector. In an embodiment, the nucleic acid vector is a DNA vector. In an embodiment, the nucleic acid vector is an RNA vector.

[0055] In exemplary aspects, a pharmaceutical composition is provided. The pharmaceutical composition comprises: i) a consensus H1 hemagglutinin polypeptide, and ii) one or more pharmaceutically acceptable carriers.

[0056] In an embodiment, a consensus H1 hemagglutinin polypeptide is comprised in a composition according to the corresponding embodiments described herein.

[0057] In an embodiment, the consensus H1 hemagglutinin polypeptide comprises the amino acid sequence

[00145] The foregoing embodiments of the amino acid sequence can be applied to the pharmaceutical compositions described herein.

[0058] In an embodiment, the consensus H1 hemagglutinin polypeptide comprises a nucleic acid sequence encoding a consensus H1 hemagglutinin polypeptide.The aforementioned nucleic acid sequence embodiments can be applied to the pharmaceutical compositions described herein.

[0059] A "pharmaceutically acceptable carrier" refers to a non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, formulation aid, or vehicle commonly used in the art to form a "pharmaceutical composition" with the therapeutic agent for administration to a subject. A pharmaceutically acceptable carrier is non-toxic to the recipient at the dosage and concentration employed and is compatible with the other ingredients of the formulation. A pharmaceutically acceptable carrier is appropriate for the formulation being used. Examples of pharmaceutically acceptable carriers include alumina; aluminum stearate; lecithin; serum proteins, such as human serum albumin, canine or other animal albumin; buffers, such as phosphate, citrate, tromethamine, or HEPES buffers; glycine; sorbic acid; potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, or magnesium trisilicate; polyvinyl pyrrolidone, a cellulose-based substance; polyethylene glycol; sucrose; mannitol; or amino acids, including but not limited to arginine.

[0060] In some embodiments, the pharmaceutically acceptable carrier has a pH of about 6.2 to about 7, about 6 to about 7.2, about 6.4 to about 6.8, about 6, or about 7 and comprises sodium phosphate and sodium chloride. In some embodiments, the pharmaceutically acceptable carrier has a pH of about 6.2 to about 7, about 6 to about 7.2, about 6, about 6.4 to about 6.8, or about 7 and comprises sodium citrate and sodium chloride.

[0061] In some embodiments, the pharmaceutically acceptable carrier comprises sodium phosphate, sodium chloride, and polysorbate 80. In some embodiments, the pharmaceutically acceptable carrier comprises sodium phosphate, sodium chloride, and polysorbate 20. In some embodiments, the pharmaceutically acceptable carrier comprises sodium citrate, sodium chloride, and polysorbate 20. In some embodiments, the pharmaceutically acceptable carrier comprises sodium citrate, sodium chloride, and polysorbate 80.

[0062] In some embodiments, the pharmaceutically acceptable carrier comprises sodium chloride at a concentration of about 100 nM to about 180 nM, about 110 nM to about 170 nM, about 120 nM to about 160 nM, about 130 nM to about 150 nM, about 140 nM, about 130 nM to about 160 nM, about 120 nM to about 150 nM, about 100 nM, about 110 nM, about 120 nM, about 130 nM, about 140 nM, about 150 nM, about 160 nM, about 170 nM, or about 180 nM.

[0063] In some embodiments, the pharmaceutically acceptable carrier comprises sodium phosphate at a concentration of about 100 nM to about 180 nM, about 110 nM to about 170 nM, about 120 nM to about 160 nM, about 130 nM to about 150 nM, about 140 nM, about 130 nM to about 160 nM, about 120 nM to about 150 nM, about 100 nM, about 110 nM, about 120 nM, about 130 nM, about 140 nM, about 150 nM, about 160 nM, about 170 nM, or about 180 nM.

[0064] In some embodiments, the pharmaceutically acceptable carrier comprises a polysorbate at a concentration of about 550 nM to about 750 nM, about 570 nM to about 730 nM, about 590 nM to about 720 nM, about 600 nM to about 700 nM, about 620 nM to about 680 nM, about 640 nM to about 660 nM, about 650 nM, about 570 nM to about 670 nM, about 550 nM to about 650 nM, about 650 nM to about 750 nM, about 630 nM to about 700 nM, or about 670 nM to about 600 nM. In some embodiments, the polysorbate is polysorbate 80. In some embodiments, the polysorbate is polysorbate 20.

[0065] In some embodiments, the pharmaceutically acceptable carrier comprises m-cresol or benzyl alcohol. In some embodiments, the concentration of m-cresol is about 0.2%, about 0.1% to about 0.3%, about 0.08% to about 0.25%, or about 0.05% to about 0.25%. In some embodiments, the concentration of benzyl alcohol is about 1%, about 0.5% to about 2%, about 0.2% to about 2.5%, about 1% to about 5%, about 0.5% to about 5%, or about 1% to about 3%.

[0066] The pharmaceutical composition can be stored in a lyophilized form; therefore, in some embodiments, the preparation method includes a lyophilization step. The lyophilized composition is then reconstituted, typically as an aqueous composition suitable for parenteral administration, and then administered to a cat. In other embodiments, particularly where the protein is highly stable to thermal and oxidative denaturation, the pharmaceutical composition can be stored as a liquid, i.e., an aqueous composition, which can be administered directly or with an appropriate diluent to a dog, cat, or horse. It can be reconstituted with sterile water for injection (WFI) and may include an antibacterial agent, such as benzyl alcohol. Therefore, the present invention provides pharmaceutical compositions in solid and liquid forms.

[0067] In an embodiment, the pharmaceutical composition is an oral formulation. In an embodiment, the oral formulation is selected from a tablet, a capsule, a suspension, an emulsion, a syrup, a colloidal dispersion, a dispersant, and an effervescent composition. In an embodiment, the oral formulation is a suspension. In an embodiment, the oral formulation is a reconstituted suspension.

[0068] In embodiments, the pharmaceutical composition is a parenteral formulation. In embodiments, the parenteral formulation is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous. In embodiments, the parenteral formulation is intravenous. In embodiments, the parenteral formulation is intraarterial. In embodiments, the parenteral formulation is intraperitoneal. In embodiments, the parenteral formulation is intrathecal. In embodiments, the parenteral formulation is intradermal. In embodiments, the parenteral formulation is epidural. In embodiments, the parenteral formulation is intraventricular. In embodiments, the parenteral formulation is intraurethral. In embodiments, the parenteral formulation is intrasternal. In embodiments, the parenteral formulation is intracranial. In embodiments, the parenteral formulation is intratumoral. In embodiments, the parenteral formulation is intramuscular. In embodiments, the parenteral formulation is subcutaneous.

[0069] In an embodiment, the pharmaceutical composition further comprises a second therapeutic agent. In an embodiment, the pharmaceutical composition is formulated as a vaccine. In an embodiment, the pharmaceutical composition is formulated as a single dose. In an embodiment, the pharmaceutical composition is formulated as a single unit dose.

[0070] In an exemplary aspect, a method for inducing an immune response against influenza A virus (IAV) in an animal is provided. The method includes administering to the animal a therapeutically effective amount of a pharmaceutical composition according to the corresponding embodiment described herein. As used herein, and when considering a pharmaceutical composition, the term "therapeutically effective amount" refers to the amount of the benefit that is desired for the animal. This amount can vary from one individual to another and will depend on many factors, including the overall physical condition of the animal and the underlying cause of the disease to be treated. The amount of the pharmaceutical composition used for treatment provides an acceptable rate of change and maintains the desired response at a beneficial level. The therapeutically effective amount of the pharmaceutical composition can be easily determined by one of ordinary skill in the art using publicly available materials and procedures.

[0071] In embodiments, the immune response comprises a cytotoxic T lymphocyte (CTL) response. In embodiments, the CTL response is an IAV-specific response.

[0072] In embodiments, the immune response comprises an IgG response. In embodiments, the IgG response is an IAV-specific response.

[0073] In embodiments, the immune response comprises a T cell response. In embodiments, the T cell response is an IAV specific response. In embodiments, the T cell response comprises CD8+ T cells.

[0074] In embodiments, the immune response comprises a T helper / memory cell response. In embodiments, the T helper / memory cell response is an IAV-specific response.

[0075] In embodiments, the immune response comprises an IFN-γ response. In embodiments, the IFN-γ response is an IAV-specific response.

[0076] In embodiments, the administration is prophylactic administration to an animal. In embodiments, the animal is a mammal. In embodiments, the animal is porcine.

[0077] In an embodiment, the IAV is an IAV H1 subtype. In an embodiment, the IAV is an IAV H3 subtype. In an embodiment, the IAV is an IAV H1N1 subtype. In an embodiment, the IAV is an IAV H1N2 subtype. In an embodiment, the IAV is an IAV H3N2 subtype.

[0078] In embodiments, the method comprises reducing the IAV viral load of the animal. In embodiments, the method comprises reducing the IAV viral shedding of the animal.

[0079] In an embodiment, the administration is oral administration. In an embodiment, the oral administration is selected from the group consisting of tablets, capsules, suspensions, emulsions, syrups, colloidal dispersions, dispersants, and effervescent compositions. In an embodiment, the oral formulation is a suspension. In an embodiment, the oral formulation is a reconstituted suspension.

[0080] In embodiments, administration is parenteral administration. In embodiments, parenteral administration is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intracerebroventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous. In embodiments, the parenteral formulation is intravenous. In embodiments, the parenteral formulation is intraarterial. In embodiments, the parenteral formulation is intraperitoneal. In embodiments, the parenteral formulation is intrathecal. In embodiments, the parenteral formulation is intradermal. In embodiments, the parenteral formulation is epidural. In embodiments, the parenteral formulation is intracerebroventricular. In embodiments, the parenteral formulation is intraurethral. In embodiments, the parenteral formulation is intrasternal. In embodiments, the parenteral formulation is intracranial. In embodiments, the parenteral formulation is intratumoral. In embodiments, the parenteral formulation is intramuscular. In embodiments, the parenteral formulation is subcutaneous.

[0081] In embodiments, the pharmaceutical composition is administered as a single dose. In embodiments, the pharmaceutical composition is administered as a single unit dose. In embodiments, the method further comprises administering to the animal a second therapeutic agent.

[0082] In an exemplary aspect, a method for preventing influenza A virus (IAV) infection in an animal is provided. The method comprises administering to the animal a therapeutically effective amount of a pharmaceutical composition according to a corresponding embodiment described herein. The aforementioned embodiments of the method for inducing an immune response can be applied to the method for preventing IAV infection described herein.

[0083] In exemplary aspects, amino acid sequences comprising a consensus H3 hemagglutinin polypeptide are provided.

[0084] In embodiments, the consensus H3 hemagglutinin polypeptide comprises SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 3.

[0085] In exemplary aspects, a nucleotide sequence comprising a nucleic acid sequence encoding a consensus H3 hemagglutinin polypeptide is provided.

[0086] In embodiments, the nucleic acid sequence comprises SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence is a modified native sequence to comprise codons optimized for expression in prokaryotic cells.

[0087] In embodiments, the consensus H3 hemagglutinin polypeptide comprises SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 3. In embodiments, the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 3.

[0088] In exemplary aspects, a recombinant polynucleotide is provided that comprises a heterologous regulatory element operably linked to a nucleic acid sequence encoding a consensus H3 hemagglutinin polypeptide.

[0089] The aforementioned embodiments of the consensus H3 hemagglutinin polypeptide can be applied to the recombinant polynucleotides described herein. The aforementioned embodiments of the nucleic acid sequences can be applied to the recombinant polynucleotides described herein. In embodiments, the heterologous regulatory element is a promoter that functions in prokaryotes.

[0090] In exemplary aspects, compositions are provided that include a consensus H3 hemagglutinin polypeptide and a carrier.

[0091] In embodiments, the consensus H3 hemagglutinin polypeptide comprises an amino acid sequence. In embodiments, the amino acid sequence comprises SEQ ID NO: 3. In embodiments, the amino acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 3. In embodiments, the amino acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 3. In embodiments, the amino acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 3. In embodiments, the amino acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 3. In embodiments, the amino acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 3. In embodiments, the amino acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 3. In embodiments, the amino acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 3. In embodiments, the amino acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 3.

[0092] In an embodiment, the consensus H3 hemagglutinin polypeptide comprises a nucleic acid sequence encoding a consensus H3 hemagglutinin polypeptide.

[0093] In embodiments, the nucleic acid sequence comprises SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 4. In embodiments, the nucleic acid sequence is a modified native sequence to comprise codons optimized for expression in prokaryotic cells.

[0094] In embodiments, the vector is a viral vector. In embodiments, the viral vector is an ORFV vector. In embodiments, the ORFV vector does not comprise ORFV121.

[0095] In an embodiment, the vector is a nucleic acid vector. In an embodiment, the nucleic acid vector is a DNA vector. In an embodiment, the nucleic acid vector is an RNA vector.

[0096] In an illustrative aspect, a pharmaceutical composition is provided. The pharmaceutical composition comprises: i) a consensus H3 hemagglutinin polypeptide, and ii) one or more pharmaceutically acceptable carriers.

[0097] In an embodiment, a consensus H3 hemagglutinin polypeptide is comprised in a composition according to the corresponding embodiments described herein.

[0098] In embodiments, the consensus H3 hemagglutinin polypeptide comprises the amino acid sequence

[00145] The foregoing embodiments of the amino acid sequence can be applied to the pharmaceutical compositions described herein.

[0099] In an embodiment, the consensus H3 hemagglutinin polypeptide comprises a nucleic acid sequence encoding a consensus H3 hemagglutinin polypeptide.The foregoing nucleic acid sequence embodiments can be applied to the pharmaceutical compositions described herein.

[0100] The foregoing embodiments relating to pharmaceutical compositions are applicable to other pharmaceutical compositions described herein.

[0101] In an exemplary aspect, a method of inducing an immune response against influenza A virus (IAV) in an animal is provided. The method comprises the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition according to the corresponding embodiments described herein.

[0102] The foregoing embodiments of methods for inducing an immune response can be applied to the other methods for inducing an immune response described herein.

[0103] In an exemplary aspect, a method of preventing influenza A virus (IAV) infection in an animal is provided. The method comprises administering to the animal a therapeutically effective amount of a pharmaceutical composition according to the corresponding embodiments described herein.

[0104] The foregoing embodiments of methods for inducing an immune response can be applied to the methods of preventing IAV infection described herein.

[0105] In an exemplary aspect, amino acid sequences comprising chimeric polypeptides are provided. The chimeric polypeptide may comprise a molecule comprising a first portion of an IAV polypeptide and a second portion of an IAV polypeptide. The first portion and the second portion may be derived from different IAV subtypes, such as H1, H3, H6, H8, H14, and H15. Any such portion may be prepared from a protein by standard biochemical methods or by expressing a polynucleotide encoding one or more portions / fragments. For example, the first portion may comprise the stalk region of a contemporary H1 or H3 IAV strain and the second portion may comprise a head region (e.g., an alien head region) derived from one of the H6, H8, H14, and H15 subtypes.

[0106] In embodiments, the chimeric polypeptide comprises a contemporary region and a head region. In embodiments, the contemporary region comprises an H1 hemagglutinin contemporary polypeptide, and the head region comprises an H6 contemporary polypeptide. In embodiments, the contemporary region comprises an H1 hemagglutinin contemporary polypeptide, and the head region comprises an H8 contemporary polypeptide. In embodiments, the contemporary region comprises an H3 hemagglutinin contemporary polypeptide, and the head region comprises an H14 contemporary polypeptide. In embodiments, the contemporary region comprises an H3 hemagglutinin contemporary polypeptide, and the head region comprises an H15 contemporary polypeptide.

[0107] In embodiments, the chimeric polypeptide comprises SEQ ID NO: 5. In embodiments, the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 5. In embodiments, the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 5. In embodiments, the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 5. In embodiments, the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 5. In embodiments, the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 5. In embodiments, the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 5. In embodiments, the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 5. In embodiments, the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 5.

[0108] In embodiments, the chimeric polypeptide comprises SEQ ID NO: 7. In embodiments, the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 7. In embodiments, the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 7. In embodiments, the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 7. In embodiments, the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 7. In embodiments, the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 7. In embodiments, the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 7. In embodiments, the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 7. In embodiments, the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 7.

[0109] In embodiments, the chimeric polypeptide comprises SEQ ID NO: 9. In embodiments, the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 9. In embodiments, the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 9. In embodiments, the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 9. In embodiments, the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 9. In embodiments, the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 9. In embodiments, the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 9. In embodiments, the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 9. In embodiments, the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 9.

[0110] In embodiments, the chimeric polypeptide comprises SEQ ID NO: 11. In embodiments, the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 11. In embodiments, the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 11. In embodiments, the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 11. In embodiments, the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 11. In embodiments, the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 11. In embodiments, the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 11. In embodiments, the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 11. In embodiments, the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 11.

[0111] In exemplary aspects, a nucleotide sequence comprising a nucleic acid sequence encoding a chimeric polypeptide is provided. In embodiments, the chimeric polypeptide comprises a contemporary region and a head region. In embodiments, the contemporary region comprises an H1 hemagglutinin contemporary polypeptide, and the head region comprises an H6 contemporary polypeptide. In embodiments, the contemporary region comprises an H1 hemagglutinin contemporary polypeptide, and the head region comprises an H8 contemporary polypeptide. In embodiments, the contemporary region comprises an H3 hemagglutinin contemporary polypeptide, and the head region comprises an H14 contemporary polypeptide. In embodiments, the contemporary region comprises an H3 hemagglutinin contemporary polypeptide, and the head region comprises an H15 contemporary polypeptide.

[0112] In embodiments, the nucleic acid sequence comprises SEQ ID NO: 6. In embodiments, the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 6. In embodiments, the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 6. In embodiments, the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 6. In embodiments, the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 6. In embodiments, the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 6. In embodiments, the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 6. In embodiments, the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 6. In embodiments, the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 6.

[0113] In embodiments, the nucleic acid sequence comprises SEQ ID NO: 8. In embodiments, the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 8. In embodiments, the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 8. In embodiments, the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 8. In embodiments, the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 8. In embodiments, the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 8. In embodiments, the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 8. In embodiments, the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 8. In embodiments, the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 8.

[0114] In embodiments, the nucleic acid sequence comprises SEQ ID NO: 10. In embodiments, the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 10. In embodiments, the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 10. In embodiments, the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 10. In embodiments, the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 10. In embodiments, the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 10. In embodiments, the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 10. In embodiments, the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 10. In embodiments, the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 10.

[0115] In embodiments, the nucleic acid sequence comprises SEQ ID NO: 12. In embodiments, the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 12. In embodiments, the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 12. In embodiments, the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 12. In embodiments, the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 12. In embodiments, the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 12. In embodiments, the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 12. In embodiments, the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 12. In embodiments, the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 12.

[0116] In an embodiment, the nucleic acid sequence is a native sequence modified to contain codons optimized for expression in prokaryotes.

[0117] The foregoing embodiments of chimeric polypeptides of amino acid sequences are applicable to the nucleotide sequences described herein.

[0118] In exemplary aspects, a recombinant polynucleotide is provided that comprises a heterologous regulatory element operably linked to a nucleic acid sequence encoding a chimeric polypeptide.

[0119] The foregoing embodiments of chimeric polypeptides of amino acid sequences are applicable to the recombinant polynucleotides described herein.

[0120] In exemplary aspects, compositions comprising a chimeric polypeptide and a vector are provided. The aforementioned embodiments of the chimeric polypeptide of the amino acid sequence can be applied to the compositions described herein. The aforementioned embodiments of the nucleic acid sequence can be applied to the compositions described herein.

[0121] In embodiments, the vector is a viral vector. In embodiments, the viral vector is an ORFV vector. In embodiments, the ORFV vector does not comprise ORFV121.

[0122] In an embodiment, the vector is a nucleic acid vector. In an embodiment, the nucleic acid vector is a DNA vector. In an embodiment, the nucleic acid vector is an RNA vector.

[0123] In an illustrative aspect, a pharmaceutical composition is provided. The pharmaceutical composition comprises: i) a chimeric polypeptide, and ii) one or more pharmaceutically acceptable carriers.

[0124] In an embodiment, the chimeric polypeptide is comprised in a composition according to the corresponding embodiment described herein.

[0125] The aforementioned embodiments of the chimeric polypeptides of the amino acid sequences can be applied to the pharmaceutical compositions described herein. The aforementioned embodiments of the nucleic acid sequences can be applied to the pharmaceutical compositions described herein.

[0126] The foregoing embodiments relating to pharmaceutical compositions are applicable to other pharmaceutical compositions described herein.

[0127] In an exemplary aspect, a method of inducing an immune response against influenza A virus (IAV) in an animal is provided. The method comprises the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition according to the corresponding embodiments described herein.

[0128] The foregoing embodiments of methods for inducing an immune response can be applied to the other methods for inducing an immune response described herein.

[0129] In an exemplary aspect, a method of preventing influenza A virus (IAV) infection in an animal is provided. The method comprises administering to the animal a therapeutically effective amount of a pharmaceutical composition according to the corresponding embodiments described herein.

[0130] The aforementioned embodiments of the methods for inducing an immune response can be applied to the methods of preventing IAV infection described herein.

[0131] The following numbered embodiments are considered and are non-limiting: 1. An amino acid sequence comprising a consensus H1 hemagglutinin polypeptide.

[0132] 2. The amino acid sequence of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises SEQ ID NO: 1.

[0133] 3. The amino acid sequence of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 1.

[0134] 4. The amino acid sequence of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 85% identical to SEQ ID NO: 1.

[0135] 5. The amino acid sequence of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 1.

[0136] 6. The amino acid sequence of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 1.

[0137] 7. The amino acid sequence of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 96% identical to SEQ ID NO: 1.

[0138] 8. The amino acid sequence of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 1.

[0139] 9. The amino acid sequence of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 98% identical to SEQ ID NO: 1.

[0140] 10. The amino acid sequence of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 99% identical to SEQ ID NO: 1.

[0141] 11. A nucleotide sequence comprising a nucleic acid sequence encoding a consensus H1 hemagglutinin polypeptide.

[0142] 12. The nucleotide according to clause 11, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 2.

[0143] 13. The nucleotide of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 2.

[0144] 14. The nucleotide of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 2.

[0145] 15. The nucleotide of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 2.

[0146] 16. The nucleotide of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 2.

[0147] 17. The nucleotide of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 2.

[0148] 18. The nucleotide of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 2.

[0149] 19. The nucleotide of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 2.

[0150] 20. The nucleotide of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 2.

[0151] 21. The nucleotide sequence of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a native sequence modified to include codons optimized for expression in prokaryotes.

[0152] 22. The nucleotide sequence of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises SEQ ID NO: 1.

[0153] 23. The nucleotide sequence of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 80% identical to that of SEQ ID NO: 1.

[0154] 24. The nucleotide sequence of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 85% identical to that of SEQ ID NO: 1.

[0155] 25. The nucleotide sequence of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 90% identical to that of SEQ ID NO: 1.

[0156] 26. The nucleotide sequence of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 95% identical to that of SEQ ID NO: 1.

[0157] 27. The nucleotide sequence of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 96% identical to that of SEQ ID NO: 1.

[0158] 28. The nucleotide sequence of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 97% identical to that of SEQ ID NO: 1.

[0159] 29. The nucleotide sequence of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 98% identical to that of SEQ ID NO: 1.

[0160] 30. The nucleotide sequence of clause 11, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 99% identical to that of SEQ ID NO: 1.

[0161] 31. A recombinant polynucleotide comprising a heterologous regulatory element operably linked to a nucleic acid sequence encoding a consensus H1 hemagglutinin polypeptide.

[0162] 32. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises SEQ ID NO: 1.

[0163] 33. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 80% identical to that of SEQ ID NO: 1.

[0164] 34. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 85% identical to that of SEQ ID NO: 1.

[0165] 35. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 90% identical to that of SEQ ID NO: 1.

[0166] 36. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 95% identical to that of SEQ ID NO: 1.

[0167] 37. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 96% identical to that of SEQ ID NO: 1.

[0168] 38. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 97% identical to that of SEQ ID NO: 1.

[0169] 39. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 98% identical to that of SEQ ID NO: 1.

[0170] 40. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence at least 99% identical to that of SEQ ID NO: 1.

[0171] 41. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the heterologous regulatory element is a promoter functional in a prokaryotic cell.

[0172] 42. A recombinant polynucleotide according to clause 31, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 2.

[0173] 43. A recombinant polynucleotide according to clause 31, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 2.

[0174] 44. A recombinant polynucleotide according to clause 31, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 2.

[0175] 45. A recombinant polynucleotide according to clause 31, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 2.

[0176] 46. ​​A recombinant polynucleotide according to clause 31, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 2.

[0177] 47. A recombinant polynucleotide according to clause 31, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 2.

[0178] 48. A recombinant polynucleotide according to clause 31, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 2.

[0179] 49. A recombinant polynucleotide according to clause 31, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 2.

[0180] 50. A recombinant polynucleotide according to clause 31, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 2.

[0181] 51. The recombinant polynucleotide of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a native sequence modified to comprise codons optimized for expression in a prokaryotic cell.

[0182] 52. A composition comprising a consensus H1 hemagglutinin polypeptide and a carrier.

[0183] 53. The composition of clause 52, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises an amino acid sequence.

[0184] 54. A composition according to clause 53, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises SEQ ID NO: 1.

[0185] 55. A composition according to clause 53, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 1.

[0186] 56. A composition according to clause 53, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 1.

[0187] 57. A composition according to clause 53, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 90% identical to that of SEQ ID NO: 1.

[0188] 58. A composition according to clause 53, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 1.

[0189] 59. A composition according to clause 53, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 1.

[0190] 60. A composition according to clause 53, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence at least 97% identical to that of SEQ ID NO: 1.

[0191] 61. A composition according to clause 53, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 98% identical to that of SEQ ID NO: 1.

[0192] 62. A composition according to clause 53, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 99% identical to that of SEQ ID NO: 1.

[0193] 63. The composition of clause 52, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a nucleic acid sequence encoding the consensus H1 hemagglutinin polypeptide.

[0194] 64. A composition according to clause 63, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 2.

[0195] 65. A composition according to clause 63, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 2.

[0196] 66. A composition according to clause 63, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 2.

[0197] 67. A composition according to clause 63, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 2.

[0198] 68. A composition according to clause 63, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 2.

[0199] 69. A composition according to clause 63, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 2.

[0200] 70. A composition according to clause 63, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 2.

[0201] 71. A composition according to clause 63, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 2.

[0202] 72. A composition according to clause 63, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 2.

[0203] 73. A composition according to clause 63, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a native sequence modified to comprise codons optimized for expression in prokaryotic cells.

[0204] 74. A composition according to clause 52, any other suitable clause, or any combination of suitable clauses, wherein the vector is a viral vector.

[0205] 75. A composition according to clause 74, any other suitable clause, or any combination of suitable clauses, wherein the viral vector is an ORFV vector.

[0206] 76. A composition according to clause 75, any other suitable clause, or any combination of suitable clauses, wherein the ORFV vector does not comprise ORFV121.

[0207] 77. A composition according to clause 52, any other suitable clause, or any combination of suitable clauses, wherein the vector is a nucleic acid vector.

[0208] 78. A composition according to clause 77, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid vector is a DNA vector.

[0209] 79. A composition according to clause 77, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid vector is an RNA vector.

[0210] 80. A pharmaceutical composition comprising: i) a consensus H1 hemagglutinin polypeptide, and ii) one or more pharmaceutically acceptable carriers.

[0211] 81. The pharmaceutical composition of clause 80, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide is comprised in the composition of any one of clauses 52 to 79.

[0212] 82. The pharmaceutical composition of clause 80, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises an amino acid sequence.

[0213] 83. A pharmaceutical composition according to clause 82, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises SEQ ID NO: 1.

[0214] 84. A pharmaceutical composition according to clause 82, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 1.

[0215] 85. A pharmaceutical composition according to clause 82, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 1.

[0216] 86. A pharmaceutical composition according to clause 82, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 1.

[0217] 87. A pharmaceutical composition according to clause 82, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 95% identical to that of SEQ ID NO: 1.

[0218] 88. A pharmaceutical composition according to clause 82, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 1.

[0219] 89. A pharmaceutical composition according to clause 82, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 1.

[0220] 90. A pharmaceutical composition according to clause 82, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 1.

[0221] 91. A pharmaceutical composition according to clause 82, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 1.

[0222] 92. The pharmaceutical composition of clause 80, any other suitable clause, or any combination of suitable clauses, wherein the consensus H1 hemagglutinin polypeptide comprises a nucleic acid sequence encoding the consensus H1 hemagglutinin polypeptide.

[0223] 93. A pharmaceutical composition according to clause 92, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 2.

[0224] 94. A pharmaceutical composition according to clause 92, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 2.

[0225] 95. A pharmaceutical composition according to clause 92, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 2.

[0226] 96. A pharmaceutical composition according to clause 92, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 2.

[0227] 97. A pharmaceutical composition according to clause 92, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 2.

[0228] 98. A pharmaceutical composition according to clause 92, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 2.

[0229] 99. A pharmaceutical composition according to clause 92, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 2.

[0230] 100. A pharmaceutical composition according to clause 92, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 2.

[0231] 101. A pharmaceutical composition according to clause 92, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 2.

[0232] 102. A pharmaceutical composition according to clause 92, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a modified native sequence to contain codons optimized for expression in prokaryotic cells.

[0233] 103. A pharmaceutical composition according to clause 80, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is an oral formulation.

[0234] 104. The pharmaceutical composition of clause 103, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is selected from tablets, capsules, suspensions, emulsions, syrups, colloidal dispersions, dispersants and effervescent compositions.

[0235] 105. A pharmaceutical composition according to clause 103, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a suspension.

[0236] 106. A pharmaceutical composition according to clause 103, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a reconstitutable suspension.

[0237] 107. A pharmaceutical composition according to clause 80, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is a parenteral formulation.

[0238] 108. The pharmaceutical composition of clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.

[0239] 109. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intravenous.

[0240] 110. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intra-arterial.

[0241] 111. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraperitoneal.

[0242] 112. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrathecal.

[0243] 113. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intradermal.

[0244] 114. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is epidural.

[0245] 115. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraventricular.

[0246] 116. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraurethral.

[0247] 117. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrasternal.

[0248] 118. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intracranial.

[0249] 119. The pharmaceutical composition of clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intratumoral.

[0250] 120. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intramuscular.

[0251] 121. A pharmaceutical composition according to clause 107, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is subcutaneous.

[0252] 122. A pharmaceutical composition according to clause 80, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition further comprises a second therapeutic agent.

[0253] 123. A pharmaceutical composition according to clause 80, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is formulated as a vaccine.

[0254] 124. A pharmaceutical composition according to clause 80, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is formulated as a single dose.

[0255] 125. The pharmaceutical composition of clause 80, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is formulated as a single unit dose.

[0256] 126. A method of inducing an immune response against influenza A virus (IAV) in an animal, the method comprising the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition according to any one of clauses 80 to 125.

[0257] 127. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises a cytotoxic T lymphocyte (CTL) response.

[0258] 128. The method of clause 127, any other suitable clause, or any combination of suitable clauses, wherein the CTL response is an IAV-specific response.

[0259] 129. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises an IgG response.

[0260] 130. The method of clause 129, any other suitable clause, or any combination of suitable clauses, wherein the IgG response is an IAV-specific response.

[0261] 131. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises a T cell response.

[0262] 132. The method of clause 131, any other suitable clause, or any combination of suitable clauses, wherein the T cell response is an IAV-specific response.

[0263] 133. The method of clause 131, any other suitable clause, or any combination of suitable clauses, wherein the T cell response comprises CD8+ T cells.

[0264] 134. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises a T helper / memory cell response.

[0265] 135. The method of clause 134, any other suitable clause, or any combination of suitable clauses, wherein the T helper / memory cell response is an IAV-specific response.

[0266] 136. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises an IFN-γ response.

[0267] 137. The method of clause 136, any other suitable clause, or any combination of suitable clauses, wherein the IFN-γ response is an IAV-specific response.

[0268] 138. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein said administration is prophylactic administration to said animal.

[0269] 139. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the animal is a mammal.

[0270] 140. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the animal is porcine.

[0271] 141. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1 subtype.

[0272] 142. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H3 subtype.

[0273] 143. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N1 subtype.

[0274] 144. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N2 subtype.

[0275] 145. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H3N2 subtype.

[0276] 146. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing the IAV viral load in the animal.

[0277] 147. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing IAV viral shedding in the animal.

[0278] 148. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the administration is oral administration.

[0279] 149. The method of clause 148, any other suitable clause, or any combination of suitable clauses, wherein the oral administration is selected from tablets, capsules, suspensions, emulsions, syrups, colloidal dispersions, dispersants and effervescent compositions.

[0280] 150. The method of clause 148, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a suspension.

[0281] 151. The method of clause 148, any other suitable clause, or any combination of suitable clauses, wherein the oral dosage form is a reconstitutable suspension.

[0282] 152. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the administration is parenteral.

[0283] 153. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral administration is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.

[0284] 154. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intravenous.

[0285] 155. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intra-arterial.

[0286] 156. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraperitoneal.

[0287] 157. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrathecal.

[0288] 158. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intradermal.

[0289] 159. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is epidural.

[0290] 160. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraventricular.

[0291] 161. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraurethral.

[0292] 162. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrasternal.

[0293] 163. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intracranial.

[0294] 164. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intratumoral.

[0295] 165. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intramuscular.

[0296] 166. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is subcutaneous.

[0297] 167. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single dose.

[0298] 168. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single unit dose.

[0299] 169. The method of clause 126, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administering to the animal a second therapeutic agent.

[0300] 170. A method of preventing influenza A virus (IAV) infection in an animal, the method comprising the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition according to any one of clauses 80 to 125.

[0301] 171. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein said administration is prophylactic administration to said animal.

[0302] 172. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the animal is a mammal.

[0303] 173. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the animal is porcine.

[0304] 174. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1 subtype.

[0305] 175. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H3 subtype.

[0306] 176. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N1 subtype.

[0307] 177. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N2 subtype.

[0308] 178. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H3N2 subtype.

[0309] 179. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing the IAV viral load in the animal.

[0310] 180. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing IAV viral shedding in the animal.

[0311] 181. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the administration is oral administration.

[0312] 182. The method of clause 181, any other suitable clause, or any combination of suitable clauses, wherein the oral administration is selected from tablets, capsules, suspensions, emulsions, syrups, colloidal dispersions, dispersants and effervescent compositions.

[0313] 183. The method of clause 181, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a suspension.

[0314] 184. The method of clause 181, any other suitable clause, or any combination of suitable clauses, wherein the oral dosage form is a reconstitutable suspension.

[0315] 185. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the administration is parenteral.

[0316] 186. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral administration is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.

[0317] 187. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intravenous.

[0318] 188. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intra-arterial.

[0319] 189. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraperitoneal.

[0320] 190. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrathecal.

[0321] 191. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intradermal.

[0322] 192. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is epidural.

[0323] 193. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraventricular.

[0324] 194. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraurethral.

[0325] 195. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrasternal.

[0326] 196. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intracranial.

[0327] 197. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intratumoral.

[0328] 198. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intramuscular.

[0329] 199. The method of clause 185, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is subcutaneous.

[0330] 200. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single dose.

[0331] 201. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single unit dose.

[0332] 202. The method of clause 170, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administering to the animal a second therapeutic agent.

[0333] 203. An amino acid sequence comprising a consensus H3 hemagglutinin polypeptide.

[0334] 204. The amino acid sequence of clause 203, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises SEQ ID NO: 3.

[0335] 205. The amino acid sequence of clause 203, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 3.

[0336] 206. The amino acid sequence of clause 203, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 3.

[0337] 207. The amino acid sequence of clause 203, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 3.

[0338] 208. The amino acid sequence of clause 203, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 3.

[0339] 209. The amino acid sequence of clause 203, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 3.

[0340] 210. The amino acid sequence of clause 203, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 3.

[0341] 211. The amino acid sequence of clause 203, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 98% identical to SEQ ID NO: 3.

[0342] 212. The amino acid sequence of clause 203, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 99% identical to SEQ ID NO: 3.

[0343] 213. A nucleotide sequence comprising a nucleic acid sequence encoding a consensus H3 hemagglutinin polypeptide.

[0344] 214. A nucleotide sequence according to clause 213, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 4.

[0345] 215. A nucleotide sequence according to clause 213, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 4.

[0346] 216. A nucleotide sequence according to clause 213, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 4.

[0347] 217. A nucleotide sequence according to clause 213, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 4.

[0348] 218. A nucleotide sequence according to clause 213, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 4.

[0349] 219. A nucleotide sequence according to clause 213, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 4.

[0350] 220. A nucleotide sequence according to clause 213, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 4.

[0351] 221. A nucleotide sequence according to clause 213, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 4.

[0352] 222. A nucleotide sequence according to clause 213, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 4.

[0353] 223. A nucleotide sequence according to clause 213, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a native sequence modified to comprise codons optimized for expression in prokaryotic cells.

[0354] 224. The nucleotide sequence of clause 213, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises SEQ ID NO: 3.

[0355] 225. The nucleotide sequence of clause 213, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 3.

[0356] 226. The nucleotide sequence of clause 213, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 3.

[0357] 227. The nucleotide sequence of clause 213, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 3.

[0358] 228. The nucleotide sequence of clause 213, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 3.

[0359] 229. The nucleotide sequence of clause 213, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 3.

[0360] 230. The nucleotide sequence of clause 213, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 3.

[0361] 231. The nucleotide sequence of clause 213, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 3.

[0362] 232. The nucleotide sequence of clause 213, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 3.

[0363] 233. A recombinant polynucleotide comprising a heterologous regulatory element operably linked to a nucleic acid sequence encoding a consensus H3 hemagglutinin polypeptide.

[0364] 234. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises SEQ ID NO: 3.

[0365] 235. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence at least 80% identical to that of SEQ ID NO: 3.

[0366] 236. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence at least 85% identical to that of SEQ ID NO: 3.

[0367] 237. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence at least 90% identical to that of SEQ ID NO: 3.

[0368] 238. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence at least 95% identical to that of SEQ ID NO: 3.

[0369] 239. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence at least 96% identical to that of SEQ ID NO: 3.

[0370] 240. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence at least 97% identical to that of SEQ ID NO: 3.

[0371] 241. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence at least 98% identical to that of SEQ ID NO: 3.

[0372] 242. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence at least 99% identical to that of SEQ ID NO: 3.

[0373] 243. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the heterologous regulatory element is a promoter functional in a prokaryotic cell.

[0374] 244. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 4.

[0375] 245. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 4.

[0376] 246. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 4.

[0377] 247. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 4.

[0378] 248. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 4.

[0379] 249. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 4.

[0380] 250. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 4.

[0381] 251. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 4.

[0382] 252. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 4.

[0383] 253. A recombinant polynucleotide according to clause 233, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a native sequence modified to comprise codons optimized for expression in a prokaryotic cell.

[0384] 254. A composition comprising a consensus H3 hemagglutinin polypeptide and a carrier.

[0385] 255. A composition according to clause 254, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises an amino acid sequence.

[0386] 256. A composition according to clause 255, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises SEQ ID NO: 3.

[0387] 257. A composition according to clause 255, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 3.

[0388] 258. A composition according to clause 255, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 3.

[0389] 259. A composition according to clause 255, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 3.

[0390] 260. A composition according to clause 255, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 3.

[0391] 261. A composition according to clause 255, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 3.

[0392] 262. A composition according to clause 255, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 3.

[0393] 263. A composition according to clause 255, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 3.

[0394] 264. A composition according to clause 255, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 3.

[0395] 265. A composition according to clause 254, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a nucleic acid sequence encoding the consensus H3 hemagglutinin polypeptide.

[0396] 266. A composition according to clause 265, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 4.

[0397] 267. A composition according to clause 265, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 4.

[0398] 268. A composition according to clause 265, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 4.

[0399] 269. A composition according to clause 265, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 4.

[0400] 270. A composition according to clause 265, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 4.

[0401] 271. A composition according to clause 265, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 4.

[0402] 272. A composition according to clause 265, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 4.

[0403] 273. A composition according to clause 265, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 4.

[0404] 274. A composition according to clause 265, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 4.

[0405] 275. A composition according to clause 265, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a modified native sequence to comprise codons optimized for expression in prokaryotic cells.

[0406] 276. A composition according to clause 254, any other suitable clause, or any combination of suitable clauses, wherein the vector is a viral vector.

[0407] 277. A composition according to clause 276, any other suitable clause, or any combination of suitable clauses, wherein the viral vector is an ORFV vector.

[0408] 278. A composition according to clause 277, any other suitable clause, or any combination of suitable clauses, wherein the ORFV vector does not comprise ORFV121.

[0409] 279. A composition according to clause 254, any other suitable clause, or any combination of suitable clauses, wherein the vector is a nucleic acid vector.

[0410] 280. A composition according to clause 279, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid vector is a DNA vector.

[0411] 281. A composition according to clause 279, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid vector is an RNA vector.

[0412] 282. A pharmaceutical composition comprising: i) a consensus H3 hemagglutinin polypeptide, and ii) one or more pharmaceutically acceptable carriers.

[0413] 283. A pharmaceutical composition according to clause 282, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide is comprised in the composition of any one of clauses 254 to 281.

[0414] 284. The pharmaceutical composition of clause 282, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises the amino acid sequence.

[0415] 285. A pharmaceutical composition according to clause 284, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises SEQ ID NO: 3.

[0416] 286. A pharmaceutical composition according to clause 284, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 3.

[0417] 287. A pharmaceutical composition according to clause 284, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 3.

[0418] 288. A pharmaceutical composition according to clause 284, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 3.

[0419] 289. A pharmaceutical composition according to clause 284, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 3.

[0420] 290. A pharmaceutical composition according to clause 284, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 3.

[0421] 291. A pharmaceutical composition according to clause 284, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 3.

[0422] 292. A pharmaceutical composition according to clause 284, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 3.

[0423] 293. A pharmaceutical composition according to clause 284, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 3.

[0424] 294. The pharmaceutical composition of clause 282, any other suitable clause, or any combination of suitable clauses, wherein the consensus H3 hemagglutinin polypeptide comprises a nucleic acid sequence encoding the consensus H3 hemagglutinin polypeptide.

[0425] 295. A pharmaceutical composition according to clause 294, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 4.

[0426] 296. A pharmaceutical composition according to clause 294, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 4.

[0427] 297. A pharmaceutical composition according to clause 294, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 4.

[0428] 298. A pharmaceutical composition according to clause 294, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 4.

[0429] 299. A pharmaceutical composition according to clause 294, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 4.

[0430] 300. A pharmaceutical composition according to clause 294, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 4.

[0431] 301. A pharmaceutical composition according to clause 294, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 4.

[0432] 302. A pharmaceutical composition according to clause 294, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 4.

[0433] 303. A pharmaceutical composition according to clause 294, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 4.

[0434] 304. A pharmaceutical composition according to clause 294, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a native sequence modified to comprise codons optimized for expression in prokaryotic cells.

[0435] 305. A pharmaceutical composition according to clause 282, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is an oral formulation.

[0436] 306. A pharmaceutical composition according to clause 305, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is selected from tablets, capsules, suspensions, emulsions, syrups, colloidal dispersions, dispersants and effervescent compositions.

[0437] 307. A pharmaceutical composition according to clause 305, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a suspension.

[0438] 308. A pharmaceutical composition according to clause 305, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a reconstitutable suspension.

[0439] 309. A pharmaceutical composition according to clause 282, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is a parenteral formulation.

[0440] 310. The pharmaceutical composition of clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.

[0441] 311. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intravenous.

[0442] 312. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intra-arterial.

[0443] 313. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraperitoneal.

[0444] 314. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrathecal.

[0445] 315. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intradermal.

[0446] 316. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is epidural.

[0447] 317. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraventricular.

[0448] 318. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraurethral.

[0449] 319. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrasternal.

[0450] 320. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intracranial.

[0451] 321. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intratumoral.

[0452] 322. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intramuscular.

[0453] 323. A pharmaceutical composition according to clause 309, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is subcutaneous.

[0454] 324. A pharmaceutical composition according to clause 282, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition further comprises a second therapeutic agent.

[0455] 325. A pharmaceutical composition according to clause 282, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is formulated as a vaccine.

[0456] 326. A pharmaceutical composition according to clause 282, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is formulated as a single dose.

[0457] 327. A pharmaceutical composition according to clause 282, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is formulated as a single unit dose.

[0458] 328. A method of inducing an immune response against influenza A virus (IAV) in an animal, the method comprising the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition according to any one of clauses 282 to 327.

[0459] 329. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises a cytotoxic T lymphocyte (CTL) response.

[0460] 330. The method of clause 329, any other suitable clause, or any combination of suitable clauses, wherein the CTL response is an IAV-specific response.

[0461] 331. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises an IgG response.

[0462] 332. The method of clause 331, any other suitable clause, or any combination of suitable clauses, wherein the IgG response is an IAV-specific response.

[0463] 333. A method according to clause 328, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises a T cell response.

[0464] 334. The method of clause 333, any other suitable clause, or any combination of suitable clauses, wherein the T cell response is an IAV-specific response.

[0465] 335. A method according to clause 333, any other suitable clause, or any combination of suitable clauses, wherein the T cell response comprises CD8+ T cells.

[0466] 336. A method according to clause 328, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises a T helper / memory cell response.

[0467] 337. The method of clause 336, any other suitable clause, or any combination of suitable clauses, wherein the T helper / memory cell response is an IAV-specific response.

[0468] 338. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises an IFN-γ response.

[0469] 339. The method of clause 338, any other suitable clause, or any combination of suitable clauses, wherein the IFN-γ response is an IAV-specific response.

[0470] 340. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein said administration is prophylactic administration to said animal.

[0471] 341. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the animal is a mammal.

[0472] 342. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the animal is porcine.

[0473] 343. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1 subtype.

[0474] 344. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H3 subtype.

[0475] 345. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N1 subtype.

[0476] 346. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N2 subtype.

[0477] 347. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the IAV is an IAV H3N2 subtype.

[0478] 348. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing the IAV viral load in the animal.

[0479] 349. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing IAV viral shedding in the animal.

[0480] 350. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the administration is oral administration.

[0481] 351. The method of clause 350, any other suitable clause, or any combination of suitable clauses, wherein the oral administration is selected from tablets, capsules, suspensions, emulsions, syrups, colloidal dispersions, dispersants and effervescent compositions.

[0482] 352. The method of clause 350, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a suspension.

[0483] 353. The method of clause 350, any other suitable clause, or any combination of suitable clauses, wherein the oral dosage form is a reconstitutable suspension.

[0484] 354. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the administration is parenteral.

[0485] 355. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral administration is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.

[0486] 356. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intravenous.

[0487] 357. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intra-arterial.

[0488] 358. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraperitoneal.

[0489] 359. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrathecal.

[0490] 360. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intradermal.

[0491] 361. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is epidural.

[0492] 362. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraventricular.

[0493] 363. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraurethral.

[0494] 364. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrasternal.

[0495] 365. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intracranial.

[0496] 366. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intratumoral.

[0497] 367. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intramuscular.

[0498] 368. The method of clause 354, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is subcutaneous.

[0499] 369. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single dose.

[0500] 370. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single unit dose.

[0501] 371. The method of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administering a second therapeutic agent to the animal.

[0502] 372. A method of preventing influenza A virus (IAV) infection in an animal, the method comprising the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition according to any one of clauses 282 to 327.

[0503] 373. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein said administration is prophylactic administration to said animal.

[0504] 374. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the animal is a mammal.

[0505] 375. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the animal is porcine.

[0506] 376. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1 subtype.

[0507] 377. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H3 subtype.

[0508] 378. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N1 subtype.

[0509] 379. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N2 subtype.

[0510] 380. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the IAV is of the IAV H3N2 subtype.

[0511] 381. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing the IAV viral load in the animal.

[0512] 382. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing IAV viral shedding in the animal.

[0513] 383. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the administration is oral administration.

[0514] 384. The method of clause 383, any other suitable clause, or any combination of suitable clauses, wherein the oral administration is selected from tablets, capsules, suspensions, emulsions, syrups, colloidal dispersions, dispersants and effervescent compositions.

[0515] 385. The method of clause 383, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a suspension.

[0516] 386. The method of clause 383, any other suitable clause, or any combination of suitable clauses, wherein the oral dosage form is a reconstitutable suspension.

[0517] 387. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the administration is parenteral.

[0518] 388. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral administration is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.

[0519] 389. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intravenous.

[0520] 390. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intra-arterial.

[0521] 391. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraperitoneal.

[0522] 392. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrathecal.

[0523] 393. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intradermal.

[0524] 394. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is epidural.

[0525] 395. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraventricular.

[0526] 396. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraurethral.

[0527] 397. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrasternal.

[0528] 398. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intracranial.

[0529] 399. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intratumoral.

[0530] 400. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intramuscular.

[0531] 401. The method of clause 387, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is subcutaneous.

[0532] 402. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single dose.

[0533] 403. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single unit dose.

[0534] 404. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administering to the animal a second therapeutic agent.

[0535] 405. An amino acid sequence comprising a chimeric polypeptide.

[0536] 406. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a umbilical region and a head region.

[0537] 407. The amino acid sequence of clause 406, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H1 hemagglutinin contemporary polypeptide and the head region comprises an H6 contemporary polypeptide.

[0538] 408. The amino acid sequence of clause 406, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H1 hemagglutinin contemporary polypeptide and the head region comprises an H8 contemporary polypeptide.

[0539] 409. The amino acid sequence of clause 406, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H3 hemagglutinin contemporary polypeptide and the head region comprises an H14 contemporary polypeptide.

[0540] 410. The amino acid sequence of clause 406, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H3 hemagglutinin contemporary polypeptide and the head region comprises an H15 contemporary polypeptide.

[0541] 411. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 5.

[0542] 412. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 5.

[0543] 413. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 5.

[0544] 414. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 5.

[0545] 415. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 5.

[0546] 416. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 5.

[0547] 417. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 5.

[0548] 418. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 5.

[0549] 419. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 5.

[0550] 420. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 7.

[0551] 421. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 7.

[0552] 422. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 7.

[0553] 423. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 7.

[0554] 424. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 7.

[0555] 425. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 7.

[0556] 426. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 7.

[0557] 427. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 7.

[0558] 428. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 7.

[0559] 429. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 9.

[0560] 430. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 9.

[0561] 431. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 9.

[0562] 432. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 9.

[0563] 433. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 9.

[0564] 434. The method of clause 405, any other suitable clause, or any combination of suitable clause amino acid sequences, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 9.

[0565] 435. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 9.

[0566] 436. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 9.

[0567] 437. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 9.

[0568] 438. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 11.

[0569] 439. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 11.

[0570] 440. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 11.

[0571] 441. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 11.

[0572] 442. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 11.

[0573] 443. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 11.

[0574] 444. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 11.

[0575] 445. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 11.

[0576] 446. The amino acid sequence of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 11.

[0577] 447. A nucleotide sequence comprising a nucleic acid sequence encoding a chimeric polypeptide.

[0578] 448. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a contemporary region and a head region.

[0579] 449. A nucleotide sequence according to clause 448, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H1 hemagglutinin contemporary polypeptide and the head region comprises an H6 contemporary polypeptide.

[0580] 450. A nucleotide sequence according to clause 448, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H1 hemagglutinin contemporary polypeptide and the head region comprises an H8 contemporary polypeptide.

[0581] 451. A nucleotide sequence according to clause 448, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H3 hemagglutinin contemporary polypeptide and the head region comprises an H14 contemporary polypeptide.

[0582] 452. A nucleotide sequence according to clause 448, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H3 hemagglutinin contemporary polypeptide and the head region comprises an H15 contemporary polypeptide.

[0583] 453. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 6.

[0584] 454. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 6.

[0585] 455. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 6.

[0586] 456. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 6.

[0587] 457. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 6.

[0588] 458. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 6.

[0589] 459. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 6.

[0590] 460. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 6.

[0591] 461. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 6.

[0592] 462. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 8.

[0593] 463. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 8.

[0594] 464. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 8.

[0595] 465. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 8.

[0596] 466. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 8.

[0597] 467. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 8.

[0598] 468. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 8.

[0599] 469. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 8.

[0600] 470. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 8.

[0601] 471. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 10.

[0602] 472. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 10.

[0603] 473. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 10.

[0604] 474. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 10.

[0605] 475. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 10.

[0606] 476. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 10.

[0607] 477. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 10.

[0608] 478. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 10.

[0609] 479. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 10.

[0610] 480. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 12.

[0611] 481. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 12.

[0612] 482. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 12.

[0613] 483. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 12.

[0614] 484. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 12.

[0615] 485. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 12.

[0616] 486. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence at least 97% identical to the sequence of SEQ ID NO: 12.

[0617] 487. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 12.

[0618] 488. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence at least 99% identical to the sequence of SEQ ID NO: 12.

[0619] 489. A nucleotide sequence according to clause 447, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a native sequence modified to comprise codons optimized for expression in a prokaryotic cell.

[0620] 490. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 5.

[0621] 491. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 5.

[0622] 492. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 5.

[0623] 493. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 5.

[0624] 494. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 5.

[0625] 495. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 5.

[0626] 496. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 5.

[0627] 497. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 5.

[0628] 498. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 5.

[0629] 499. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 7.

[0630] 500. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 7.

[0631] 501. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 7.

[0632] 502. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 7.

[0633] 503. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 7.

[0634] 504. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 7.

[0635] 505. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 7.

[0636] 506. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 7.

[0637] 507. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 7.

[0638] 508. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 9.

[0639] 509. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 9.

[0640] 510. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 9.

[0641] 511. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 9.

[0642] 512. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 9.

[0643] 513. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 9.

[0644] 514. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 9.

[0645] 515. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 9.

[0646] 516. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 9.

[0647] 517. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 11.

[0648] 518. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 11.

[0649] 519. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 11.

[0650] 520. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 11.

[0651] 521. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 11.

[0652] 522. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 11.

[0653] 523. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 11.

[0654] 524. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 11.

[0655] 525. The nucleotide sequence of clause 447, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 11.

[0656] 526. A recombinant polynucleotide comprising a heterologous regulatory element operably linked to a nucleic acid sequence encoding a chimeric polypeptide.

[0657] 527. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a contemporary region and a head region.

[0658] 528. A recombinant polynucleotide according to clause 527, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H1 hemagglutinin contemporary polypeptide and the head region comprises an H6 contemporary polypeptide.

[0659] 529. A recombinant polynucleotide according to clause 527, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H1 hemagglutinin contemporary polypeptide and the head region comprises an H8 contemporary polypeptide.

[0660] 530. A recombinant polynucleotide according to clause 527, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H3 hemagglutinin contemporary polypeptide and the head region comprises an H14 contemporary polypeptide.

[0661] 531. A recombinant polynucleotide according to clause 527, any other suitable clause, or any combination of suitable clauses, wherein the contemporary region comprises an H3 hemagglutinin contemporary polypeptide and the head region comprises an H15 contemporary polypeptide.

[0662] 532. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 5.

[0663] 533. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 5.

[0664] 534. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 5.

[0665] 535. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 5.

[0666] 536. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 5.

[0667] 537. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 5.

[0668] 538. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 5.

[0669] 539. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 5.

[0670] 540. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 5.

[0671] 541. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 7.

[0672] 542. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 7.

[0673] 543. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 7.

[0674] 544. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 7.

[0675] 545. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 7.

[0676] 546. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 7.

[0677] 547. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 7.

[0678] 548. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 7.

[0679] 549. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 7.

[0680] 550. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 9.

[0681] 551. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 9.

[0682] 552. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 9.

[0683] 553. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 9.

[0684] 554. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 9.

[0685] 555. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 9.

[0686] 556. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 9.

[0687] 557. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 9.

[0688] 558. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 9.

[0689] 559. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 11.

[0690] 560. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 11.

[0691] 561. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 11.

[0692] 562. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 11.

[0693] 563. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 11.

[0694] 564. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 11.

[0695] 565. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 11.

[0696] 566. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 11.

[0697] 567. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 11.

[0698] 568. A recombinant polynucleotide according to clause 526, any other suitable clause, or any combination of suitable clauses, wherein the heterologous regulatory element is a promoter functional in a prokaryotic cell.

[0699] 569. A composition comprising a chimeric polypeptide and a carrier.

[0700] 570. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 5.

[0701] 571. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 5.

[0702] 572. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 5.

[0703] 573. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 5.

[0704] 574. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 5.

[0705] 575. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 5.

[0706] 576. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 5.

[0707] 577. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 5.

[0708] 578. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 5.

[0709] 579. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 7.

[0710] 580. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 7.

[0711] 581. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 7.

[0712] 582. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 7.

[0713] 583. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 7.

[0714] 584. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 7.

[0715] 585. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 7.

[0716] 586. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 7.

[0717] 587. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 7.

[0718] 588. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 9.

[0719] 589. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 9.

[0720] 590. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 9.

[0721] 591. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 9.

[0722] 592. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 9.

[0723] 593. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 9.

[0724] 594. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 9.

[0725] 595. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 9.

[0726] 596. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 9.

[0727] 597. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 11.

[0728] 598. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 11.

[0729] 599. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 11.

[0730] 600. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 11.

[0731] 601. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 11.

[0732] 602. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 11.

[0733] 603. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 11.

[0734] 604. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 11.

[0735] 605. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 11.

[0736] 606. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a nucleic acid sequence encoding the chimeric polypeptide.

[0737] 607. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 6.

[0738] 608. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 6.

[0739] 609. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 6.

[0740] 610. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 6.

[0741] 611. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 6.

[0742] 612. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 6.

[0743] 613. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 6.

[0744] 614. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 6.

[0745] 615. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 6.

[0746] 616. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 8.

[0747] 617. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 8.

[0748] 618. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 8.

[0749] 619. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 8.

[0750] 620. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 8.

[0751] 621. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 8.

[0752] 622. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 8.

[0753] 623. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 8.

[0754] 624. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 8.

[0755] 625. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 10.

[0756] 626. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 10.

[0757] 627. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 10.

[0758] 628. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 10.

[0759] 629. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 10.

[0760] 630. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 10.

[0761] 631. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 10.

[0762] 632. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 10.

[0763] 633. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 10.

[0764] 634. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 12.

[0765] 635. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 12.

[0766] 636. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 12.

[0767] 637. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 12.

[0768] 638. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 12.

[0769] 639. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 12.

[0770] 640. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 12.

[0771] 641. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 12.

[0772] 642. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 12.

[0773] 643. A composition according to clause 606, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a modified native sequence to comprise codons optimized for expression in a prokaryotic cell.

[0774] 644. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the vector is a viral vector.

[0775] 645. A composition according to clause 644, any other suitable clause, or any combination of suitable clauses, wherein the viral vector is an ORFV vector.

[0776] 646. A composition according to clause 645, any other suitable clause, or any combination of suitable clauses, wherein the ORFV vector does not comprise ORFV121.

[0777] 647. A composition according to clause 569, any other suitable clause, or any combination of suitable clauses, wherein the vector is a nucleic acid vector.

[0778] 648. A composition according to clause 647, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid vector is a DNA vector.

[0779] 649. A composition according to clause 647, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid vector is an RNA vector.

[0780] 650. A pharmaceutical composition comprising: i) a chimeric polypeptide, and ii) one or more pharmaceutically acceptable carriers.

[0781] 651. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide is comprised in the composition of any one of clauses 569 to 649.

[0782] 652. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 5.

[0783] 653. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 5.

[0784] 654. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 5.

[0785] 655. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 5.

[0786] 656. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 5.

[0787] 657. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 5.

[0788] 658. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 5.

[0789] 659. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 5.

[0790] 660. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 5.

[0791] 661. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 7.

[0792] 662. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 7.

[0793] 663. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 7.

[0794] 664. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 7.

[0795] 665. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 7.

[0796] 666. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 7.

[0797] 667. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 7.

[0798] 668. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 7.

[0799] 669. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 7.

[0800] 670. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 9.

[0801] 671. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 9.

[0802] 672. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 9.

[0803] 673. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 9.

[0804] 674. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 9.

[0805] 675. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 9.

[0806] 676. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 9.

[0807] 677. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 9.

[0808] 678. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 9.

[0809] 679. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises SEQ ID NO: 11.

[0810] 680. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 11.

[0811] 681. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 11.

[0812] 682. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 11.

[0813] 683. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 11.

[0814] 684. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 11.

[0815] 685. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 11.

[0816] 686. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 11.

[0817] 687. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 11.

[0818] 688. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the chimeric polypeptide comprises a nucleic acid sequence encoding the chimeric polypeptide.

[0819] 689. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 6.

[0820] 690. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 6.

[0821] 691. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 6.

[0822] 692. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 6.

[0823] 693. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 6.

[0824] 694. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 6.

[0825] 695. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 6.

[0826] 696. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 6.

[0827] 697. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 6.

[0828] 698. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 8.

[0829] 699. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 8.

[0830] 700. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 8.

[0831] 701. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 8.

[0832] 702. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 8.

[0833] 703. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 8.

[0834] 704. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 8.

[0835] 705. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 8.

[0836] 706. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 8.

[0837] 707. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 10.

[0838] 708. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 10.

[0839] 709. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 10.

[0840] 710. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 10.

[0841] 711. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 10.

[0842] 712. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 10.

[0843] 713. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 10.

[0844] 714. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 10.

[0845] 715. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 10.

[0846] 716. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises SEQ ID NO: 12.

[0847] 717. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 80% identical to the sequence of SEQ ID NO: 12.

[0848] 718. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 85% identical to the sequence of SEQ ID NO: 12.

[0849] 719. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO: 12.

[0850] 720. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 12.

[0851] 721. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 96% identical to the sequence of SEQ ID NO: 12.

[0852] 722. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 97% identical to the sequence of SEQ ID NO: 12.

[0853] 723. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 98% identical to the sequence of SEQ ID NO: 12.

[0854] 724. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 12.

[0855] 725. A pharmaceutical composition according to clause 688, any other suitable clause, or any combination of suitable clauses, wherein the nucleic acid sequence is a native sequence modified to comprise codons optimized for expression in prokaryotic cells.

[0856] 726. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is an oral formulation.

[0857] 727. A pharmaceutical composition according to clause 726, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is selected from tablets, capsules, suspensions, emulsions, syrups, colloidal dispersions, dispersants and effervescent compositions.

[0858] 728. A pharmaceutical composition according to clause 726, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a suspension.

[0859] 729. A pharmaceutical composition according to clause 726, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a reconstitutable suspension.

[0860] 730. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is a parenteral formulation.

[0861] 731. The pharmaceutical composition of clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.

[0862] 732. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intravenous.

[0863] 733. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intra-arterial.

[0864] 734. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraperitoneal.

[0865] 735. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrathecal.

[0866] 736. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intradermal.

[0867] 737. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is epidural.

[0868] 738. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraventricular.

[0869] 739. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraurethral.

[0870] 740. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrasternal.

[0871] 741. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intracranial.

[0872] 742. The pharmaceutical composition of clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intratumoral.

[0873] 743. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intramuscular.

[0874] 744. A pharmaceutical composition according to clause 730, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is subcutaneous.

[0875] 745. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition further comprises a second therapeutic agent.

[0876] 746. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is formulated as a vaccine.

[0877] 747. A pharmaceutical composition according to clause 650, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is formulated as a single dose.

[0878] 748. The pharmaceutical composition of clause 650, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is formulated as a single unit dose.

[0879] 749. A method of inducing an immune response against influenza A virus (IAV) in an animal, the method comprising the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition according to any one of clauses 650 to 748.

[0880] 750. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises a cytotoxic T lymphocyte (CTL) response.

[0881] 751. The method of clause 750, any other suitable clause, or any combination of suitable clauses, wherein the CTL response is an IAV-specific response.

[0882] 752. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises an IgG response.

[0883] 753. The method of clause 752, any other suitable clause, or any combination of suitable clauses, wherein the IgG response is an IAV-specific response.

[0884] 754. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises a T cell response.

[0885] 755. The method of clause 754, any other suitable clause, or any combination of suitable clauses, wherein the T cell response is an IAV-specific response.

[0886] 756. A method according to clause 754, any other suitable clause, or any combination of suitable clauses, wherein the T cell response comprises CD8+ T cells.

[0887] 757. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises a T helper / memory cell response.

[0888] 758. The method of clause 757, any other suitable clause, or any combination of suitable clauses, wherein the T helper / memory cell response is an IAV-specific response.

[0889] 759. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the immune response comprises an IFN-γ response.

[0890] 760. The method of clause 759, any other suitable clause, or any combination of suitable clauses, wherein the IFN-γ response is an IAV-specific response.

[0891] 761. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein said administration is prophylactic administration to said animal.

[0892] 762. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the animal is a mammal.

[0893] 763. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the animal is porcine.

[0894] 764. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1 subtype.

[0895] 765. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H3 subtype.

[0896] 766. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N1 subtype.

[0897] 767. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N2 subtype.

[0898] 768. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H3N2 subtype.

[0899] 769. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing the IAV viral load in the animal.

[0900] 770. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing IAV viral shedding in the animal.

[0901] 771. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the administration is oral administration.

[0902] 772. The method of clause 771, any other suitable clause, or any combination of suitable clauses, wherein the oral administration is selected from tablets, capsules, suspensions, emulsions, syrups, colloidal dispersions, dispersants and effervescent compositions.

[0903] 773. The method of clause 771, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a suspension.

[0904] 774. The method of clause 771, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a reconstitutable suspension.

[0905] 775. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the administration is parenteral.

[0906] 776. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral administration is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.

[0907] 777. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intravenous.

[0908] 778. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intra-arterial.

[0909] 779. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraperitoneal.

[0910] 780. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrathecal.

[0911] 781. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intradermal.

[0912] 782. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is epidural.

[0913] 783. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraventricular.

[0914] 784. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraurethral.

[0915] 785. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrasternal.

[0916] 786. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intracranial.

[0917] 787. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intratumoral.

[0918] 788. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intramuscular.

[0919] 789. The method of clause 775, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is subcutaneous.

[0920] 790. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single dose.

[0921] 791. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single unit dose.

[0922] 792. The method of clause 749, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administering a second therapeutic agent to the animal.

[0923] 793. A method of preventing influenza A virus (IAV) infection in an animal, the method comprising the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition according to any one of clauses 650 to 748.

[0924] 794. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein said administration is prophylactic administration to said animal.

[0925] 795. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the animal is a mammal.

[0926] 796. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the animal is porcine.

[0927] 797. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1 subtype.

[0928] 798. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the IAV is an IAV H3 subtype.

[0929] 799. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N1 subtype.

[0930] 800. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H1N2 subtype.

[0931] 801. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the IAV is IAV H3N2 subtype.

[0932] 802. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing the IAV viral load in the animal.

[0933] 803. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the method comprises reducing IAV viral shedding in the animal.

[0934] 804. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the administration is oral administration.

[0935] 805. The method of clause 804, any other suitable clause, or any combination of suitable clauses, wherein the oral administration is selected from tablets, capsules, suspensions, emulsions, syrups, colloidal dispersions, dispersants, and effervescent compositions.

[0936] 806. The method of clause 804, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a suspension.

[0937] 807. The method of clause 804, any other suitable clause, or any combination of suitable clauses, wherein the oral formulation is a reconstitutable suspension.

[0938] 808. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the administration is parenteral.

[0939] 809. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral administration is selected from intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.

[0940] 810. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intravenous.

[0941] 811. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intra-arterial.

[0942] 812. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraperitoneal.

[0943] 813. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrathecal.

[0944] 814. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intradermal.

[0945] 815. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is epidural.

[0946] 816. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraventricular.

[0947] 817. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intraurethral.

[0948] 818. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intrasternal.

[0949] 819. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intracranial.

[0950] 820. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intratumoral.

[0951] 821. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is intramuscular.

[0952] 822. The method of clause 808, any other suitable clause, or any combination of suitable clauses, wherein the parenteral formulation is subcutaneous.

[0953] 823. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single dose.

[0954] 824. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the pharmaceutical composition is administered as a single unit dose.

[0955] 825. The method of clause 793, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administering a second therapeutic agent to the animal.

[0956] Example

[0957] Example 1

[0958] Experimental methods and materials

[0959] Cells and viruses

[0960] Primary ovine fetal turbinate (OFTu), porcine turbinate cells (STU), and Madin-Darby canine kidney (MDCK) cells were cultured in minimum essential medium (MEM) or Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS, 2 mM L-glutamine, and containing 1 mL of penicillin (100 U / ml), streptomycin (100 μg / ml), and gentamicin (50 μg / ml) at 37°C and 5% CO2. ORFV strain IA82 (OV-IA82) (provided by Dr. Daniel Rock at the University of Illinois at Urbana-Champaign) was used as the parental virus to construct a recombinant ORFV expressing the consensus H1 glycoprotein (HA) of swine influenza A virus (IAV-S). Swine influenza virus H1N1 A / Swine / OH / 24366 / 2007 (provided by Dr. Gourapura's laboratory at OSU) (H1N1 virus, belonging to the gamma clade) and pandemic H1N1 A / California / 04 / 2009 (obtained from the National Veterinary Services Laboratory [NVSL], Ames, Iowa) (belonging to the "new epidemic" (npdm) clade) were used for viral challenge and as antigens for whole virus ELISA. Both H1N1 viruses were propagated using MDCK cells in DMEM containing TPCK-treated trypsin (2 μg / ml) and 25 mM HEPES buffer. All IAV-S isolates used for cross-reactivity assays were obtained from NVSL. These isolates include: A / Swine / Iowa / A024852 / 2020 (γ branch), A / Swine / South Dakota / A02156993 / 2018 (γ-2-β-like branch), A / Swine / Missouri / A02479312 / 2020 (npdm branch), A / Swine / Michigan / A02524810 / 2020 (npdm branch), A / Swine / Texas / A02245632 / 2020 (β branch), A / Swine / Oklahoma / A02245707 / 2020 (β branch), A / Swine / Minnesota / A01785306 / 2017 (α branch), A / Swine / Iowa / A02479151 / 2020 (npdm branch), A / Swine / South Dakota / A02156993 / 2018 (γ-2-β-like branch), A / Swine / Missouri / A02479312 / 2020 (npdm branch), A / Swine / Michigan / A02524810 / 2020 (npdm branch), (δ1 branch), A / Swine / Oklahoma / A02214419 / 2017 (δ1 branch), and A / Swine / South Dakota / A02524887 / 2020 (γ branch). More information about these isolates is shown in Table 2.

[0961] Table 2. Information on the IAV-S isolates used in the ELISA, including percent nucleotide (nt) pairwise identity and percent amino acid (aa) pairwise identity with the conH1 sequence.

[0962]

[0963] Table 2 (continued)

[0964] Table 2 (continued)

[0965] Table 2 (continued)

[0966] Construction of recombinant plasmid

[0967] The consensus full-length coding sequence of HA from the H1 subtype (conH1) was generated in silico after analyzing and aligning a final set of 63 IAV-S sequences isolated from clinically diagnosed cases at the South Dakota Animal Disease Research and Diagnostic Laboratory from 2014 to 2017 (Table 3). The restriction endonuclease sites required for insertion into the ORFV121 locus of the ORFV genome and the early poxvirus transcription termination signal (TTTTTNT) were removed from the conH1 sequence by silent nucleotide substitutions. The flag tag epitope coding sequence was added to the 5' end of the HA consensus sequence under the control of the vaccinia virus late I1L promoter. Finally, coding sequences for HindIII and SalI restriction sites were added to the 5' and 3' ends of conH1. A full-length DNA fragment containing the conH1 coding sequence under the control of the I1L promoter and the Flag tag epitope was chemically synthesized (GenScript®, Piscataway, NJ) and subcloned into the poxvirus pUC57-ORFVΔ121-loxP-EGFP transfer vector described previously. Correct cloning of conH1 was confirmed by restriction enzyme analysis using a 1% agarose gel.

[0968] Table 3. The set of 63 HA sequences used for nucleotide alignment to generate the conH1 sequences listed above.

[0969] Generation and characterization of recombinant viruses

[0970] As previously described, the full-length conH1 sequence was inserted into the ORFV121 locus of the ORFV genome by homologous recombination between the parental ORFV strain IA82 and the recombinant expression cassette pUC57-ORFVΔ121-loxP-EGFP. The presence of conH1 and the absence of ORFV121 sequences in the purified recombinant virus were confirmed by PCR screening. PCR amplification of conH1 was performed using two pairs of internal primers: IAV-S-conH1-Fw-5'-ACTGCAAGCTTTATTTAAAAGTTGTTTGGTGAACTTAAATGGACTACAAAGACGATGAGACAAGAAAG-3' (SEQ ID NO: 13) and IAV-S-conH1-Rv-5'-GAGGTGTCGACTTAAATACATATTCTACACTGTAAAGAC-3' (SEQ ID. NO: 14) (1774 bp), and for ORFV121 (401 bp) amplification: ORF121-int-Fw-5'-CCTCGGAAAAGAGCAGACAC-3' (SEQ ID. NO: 15) and ORF121-int-Rv-5'-CTTCATCAGGCAGTCGTTCA-3' (SEQ ID. NO: 16), followed by analysis of PCR amplicons by electrophoresis using 1% agarose gel. The insertion and integrity of full-length conH1 and the identity and integrity of ORFVA121 were further confirmed by sequencing with the Illumina Mi-Seq sequencing platform (Illumina, San Diego, CA) using the Nextera XT DNA library preparation kit (Illumina, San Diego, CA).

[0971] ORFV Δ121 in vitro expression of conH1 by recombinant conH1 virus

[0972] The expression of conH1 by recombinant viruses was assessed by indirect immunofluorescence assay (IFA), flow cytometry (FC), and western blotting (WB) as described previously. Briefly, for IFA and FC, the expression of conH1 by recombinant viruses was assessed by using an anti-Flag tag epitope monoclonal mouse antibody (Genscript®, Piscataway, NJ) in the presence of ORFV. Δ121 In vitro expression of heterologous proteins was assessed in permeabilized and non-permeabilized OFTu cells using conH1 recombinant virus.

[0973] For IFA, goat anti-mouse IgG monoclonal antibody DyLight® 594 (Bethyl Laboratories Inc., Montgomery, TX) was used as the secondary conjugated antibody at 1: 300. All antibody dilutions were prepared in PBS containing 1% bovine serum albumin (BSA), and cells were observed under a fluorescence microscope.

[0974] For FC, antibodies were diluted in Perm / Wash 1x solution for permeabilized cells or in FACS buffer for non-permeabilized cells. Negative control wells received only FACS buffer without any antibody. After incubation with the primary antibody for 30 minutes, cells were washed and probed with Alexa fluor 594 goat anti-mouse antibody (INVI-trogen, catalog number: A11005) at 1:300. After incubation for 30 minutes, cells were washed, resuspended in 150 μl of PBS 1x, and stored in the dark at 4°C before data acquisition using an Attune NxT flow cytometer. Gating was adjusted based on mock-infected cells. Gating and data analysis were performed using FlowJo software (version 10.8.2, Tree Star Inc., Ashland, OR, USA).

[0975] Western blotting was also used to assess the expression of ORFV Δ121 conH1 recombinant virus expressing conH1. Briefly, OFTu cells cultured in 6-well plates were infected with ORFV Δ121 The conH1 recombinant virus was infected with a high MOI (MOI = 10) and collected at 12 hpi, 24 hpi, 48 hpi and 72 hpi. Uninfected OFTu cells were used as negative controls. Cells were lysed and Western blotted as described above. The constitutively expressed protein β-actin was selected as a loading control for quantitative Western blotting. Therefore, the membrane was incubated with a monoclonal antibody against β-actin (Santa Cruz Biotechnology Inc., Santa Cruz, CA) at a dilution of 1:1000 at 37°C for 2 hours. After washing, the membrane was incubated again with a secondary antibody labeled with IRDye 800CW at 37°C for 1 hour, washed, and developed by a ChemiDoc MP imaging system (Bio-Rad, Hercules, CA).

[0976] ORFV in cell culture Δ121 Stability of conH1 recombinant virus

[0977] Evaluation of ORFV insertion after serial passage at 1 MOI in OFTu cells (up to 10 passages) Δ121The stability of the conH1 gene in the ORFV121 locus of the conH1 genome was determined. After 10 passages, the conH1 gene was inoculated with ORFV121 at about 1 MOI at passage 1, 5, or 10. Δ121 OFTu cells were infected with conH1 recombinant virus in 24-well plates and incubated for 24 hpi. The cells were then fixed, permeabilized, and stained as described above, except that a goat anti-mouse IgG monoclonal antibody conjugate, DyLight® 488, was used as the secondary antibody.

[0978] ORFV Δ121 Cross-reactivity of conH1 against different swine H1N1 antisera

[0979] ORFV was detected in vitro Δ121 Cross-reactivity of conH1 recombinant virus with sera from pigs infected with three different H1N1 IAV-S strains. OFTu cells were plated in 24-well plates and infected with ORFV Δ121 The conH1 recombinant virus infects 48 hpi with 0.1 MOI. After incubation, the cells are fixed, permeabilized with 0.2% Triton X-100-PBS, and incubated with serial dilutions of H1N1 antisera developed for A / Sw / CA / 04 / 2009 (npdm branch), A / Sw / IL / 00685 / 2005 (δ2 branch) or A / Sw / KY / 02086 / 2008 (β branch) strains. The hemagglutination inhibition (HI) titer of these antisera for homologous viruses is respectively 1: 80, 1: 160 and 1: 80. After incubation for 1 h, the plate is washed with PBS (3 times), and incubated with anti-swine IgG secondary antibodies (DyLight® 488 conjugates), and further observed under a fluorescence microscope.

[0980] Replication dynamics

[0981] In vitro evaluation of ORFV in OFTu and primary porcine turbinate (STu) cells cultured in 12-well plates Δ121 Replication characteristics of conH1 recombinant virus. ORFV in OFTu cells Δ121 Multi-step (MOI = 0.1) and single-step (MOI = 10) growth curves of conH1. Cells were harvested at 6 hpi, 12 hpi, 24 hpi, 48 hpi, and 72 hpi. Uninfected OFTu and STu were used as controls (0 hpi). Viral titers were determined at each time point using the Spearman and Karber method and expressed as the median tissue culture infectious dose (TCID 50 ) / ml.

[0982] ORFV for evaluation Δ121 ConH1 vaccination of piglets protects against different swine influenza viruses

[0983] Animal experiments

[0984] Evaluation of ORFV in pigs Δ121 The immunogenicity and protective effect of conH1 recombinant virus against different strains. The viruses used for the challenge were A / Sw / OH / 24366 / 2007 (H1N1) (OH07) and A / California / 04 / 2009 (H1N1) (CA09) strains. Forty-one 3-week-old pigs (specific pathogen-free, IAV seronegative) were randomly divided into the following 5 experimental groups: Group 1, sham immunization / mock challenge (n=6); Group 2, ORFV Δ121 conH1 immunization / OH07 challenge (n=9); Group 3, ORFV Δ121 conH1 immunization / CA09 challenge (n = 9), Group 4, sham immunization / CA09 challenge (n = 9), and Group 5, sham immunization / OH07 challenge (n = 8) (Table 4).

[0985] Table 4. Experimental design of animal immunization studies DPV = days after vaccination Intramuscular (IM) injection of 2 ml containing 10 7.5 All immunizations were performed with virus suspensions in 2 ml of MEM or TCID50 / ml. Animals from Groups 1, 4, and 5 were mock-immunized with MEM. Animals from Groups 2 and 3 were immunized with ORFV. Δ121 All animals were immunized on day 0 and received a booster vaccination on day 21 after vaccination (DPV21). On DPV40, pigs from groups 2 and 5 were vaccinated with a viral suspension of OH07 virus intranasally and intratracheally (1 × 10 7 Similarly, on DPV40, pigs from Groups 3 and 4 were challenged with CA09 virus suspension intranasally and intratracheally (1×10 7 TCID50 / ml) challenge.

[0986] Animals were monitored daily. Serum samples and nasal swabs were collected on days 0 and 21 after vaccination (DPV0 and DPV21) and on days 0, 2, 4, and 6 after challenge (DPC0, DPC2, DPC4, and DPC6). All animals were handled in accordance with the Animal Welfare Act, and animal vaccination and challenge studies were conducted at The Ohio State University (OSU), following guidelines and protocols approved by the Institutional Animal Care and Use Committee (IACUC approval number 2017R00000016).

[0987] Humoral responses against different IAV-S viruses

[0988] The virus was assessed by whole virus ELISA in serum and bronchoalveolar lavage fluid using ORFV. Δ121 IAV-S-specific IgG immune responses induced by conH1 immunization. A panel of 12 WIV antigens for IgG-ELISA was prepared as described previously with some modifications. Briefly, viral culture supernatants were subjected to 30% sucrose cushion gradient ultracentrifugation at 17,000 RP for 1.5 h using an Optima-L 100K ultracentrifuge (Beckman Coulter). The viral pellet was resuspended in DMEM and heat-inactivated in a 56°C water bath for 30 min. The optimal coating antigen concentration and dilution of the secondary antibody were determined by checkerboard titration.

[0989] To detect total IAV-S-specific IgG, Immulon 1B ELISA plates (ThermoFisher Scientific, catalog number 3355) were coated in duplicate with 250 ng / well of concentrated, heat-inactivated IAV-S virions in bicarbonate / carbonate coating buffer (15 mM sodium carbonate, 35 mM sodium bicarbonate, pH 9.6) in a 100 μL volume. After incubation at 37°C for 1.5 hours, the plates were washed three times with PBS-T (1× PBS containing 0.5% Tween-20) and blocked overnight at 4°C with 200 μL / well of blocking solution (5% milk in PBS-T). The blocking reagent was removed, and the plates were washed three times with PBS-T. Test and control serum samples were diluted 1:50 in PBS-T 5% nonfat dry milk, and 100 μL of the diluted samples were added to paired coated and uncoated control wells and incubated at room temperature for 1 hour. Unbound antibodies were washed with PBS-T (3 times) and the plates were incubated with a biotinylated secondary antibody against porcine IgG (Bethyl Laboratories, catalog number: A100-104, TX) diluted (1:4000) in blocking buffer at room temperature for 1 hour, then washed (3 times) and incubated with streptavidin-HRP conjugate (Thermo Scientific, catalog number: 21136, Pierce, Rockford, IL) diluted (1:4000) in blocking solution at room temperature for 1 hour. The streptavidin-HRP conjugate reaction was developed using 3,3',5,5'-tetramethylbenzidine substrate (TMB) (KPL, catalog number: 5120-0047, Gaithersburg, MA). Finally, the colorimetric reaction was stopped by adding 100 μL of 1N HCl solution to each well. Optical density (OD) values ​​were measured at 450 nm using a microplate reader (Tecan, Mannedorf, Switzerland). The OD value of each test and control sample was normalized to the uncoated OD value.All assay formats were pre-optimized using serum samples from animals of known serological status.

[0990] Virus titration

[0991] Infectious IAV-S titers in swabs and tissue samples were determined by the Spearman and Karber method and expressed as the median tissue culture infectious dose (TCID 50) / ml. Briefly, 10-fold serial dilutions of the samples were transferred to 96-well plates pre-seeded with MDCK cells for 24 hours and incubated at 37°C for 1 hour. After 1 hour of adsorption, the inoculum was removed, and fresh DMEM containing 2 μg / ml TPCK-treated trypsin was added to the cells. After incubation at 37°C for 48 hours, the cells were fixed with 3.7% formaldehyde, washed three times with PBS, and permeabilized with 0.2% PBS-Triton X-100 for 10 minutes at room temperature. As described in "Immunofluorescence" in this section, virus-positive MDCK cells were detected by immunofluorescence assay using a mouse monoclonal antibody (mAb) targeting the conserved nucleoprotein of influenza virus (IAV-NP HB-65462 mAb; a gift from Drs. Eric Nelson and Steve Lawson of SDSU) at a dilution of 1:500, followed by incubation with goat anti-mouse IgG monoclonal antibody DyLight® 488 (Bethyl Laboratories Inc., Montgomery, TX) at a dilution of 1:350. Virus titer was defined as the reciprocal of the highest dilution of virus at which the appearance of fluorescent foci indicated infection / replication. Appropriate positive and negative control samples were included in all plates.

[0992] Real-time reverse transcriptase PCR (rRT-PCR)

[0993] Viral shedding in nasal secretions and viral load in the lungs were assessed by rRT-PCR. Viral nucleic acids were extracted from nasal swabs and lung tissue homogenates. Lung tissue from euthanized pigs was collected and lung lysates were prepared in serum-free DMEM. About 2-5 g of lung tissue from a single pig was minced and homogenized for 2 minutes using a Stomacher 400 laboratory mixer (Seward, Long Island, NY), clarified by centrifugation, and the supernatant was collected, aliquoted, and frozen at -80 ° C before use. All RNA extractions were performed using the MagMAX Core extraction kit (Thermo Fisher, Waltham, MA, USA) and the automated KingfisherFlex nucleic acid extractor (Thermo Fisher, Waltham, MA, USA) according to the manufacturer's recommendations. The presence of IAV-S RNA was assessed using an RNA-to-Ct one-step kit (Applied Biosystems) and custom-designed primers and probes for the conserved NP gene (Prime-Time qPCR probe detection system, Integrated DNA Technologies, USA). Amplification and detection were performed using a CFX96 Touch Real-Time PCR Detection System (BioRad) under the following conditions: reverse transcription at 48°C for 10 min, polymerase activation at 95°C for 10 min, and 40 cycles of denaturation at 95°C for 15 s and annealing and extension at 60°C for 60 s. 5.25 TCID 50 / ml and 10 5.38 TCID 50 / ml of OH / 07 or CA / 09 virus suspension from 10 -1 to 10 -8 A standard curve was established using 10-fold serial dilutions of 10 μg / mL. Relative viral genome copy number was calculated based on a standard curve derived from a four-parameter logistic regression analysis and determined as log10 genome copies / mL using CFX Maestro software (Bio-Rad). Positive and negative amplification controls were run alongside the test samples.

[0994] Cellular immune response

[0995] On the day of necropsy (DPC6), heparinized blood was collected for isolation of peripheral blood mononuclear cells (PBMCs), while bronchoalveolar lavage (BAL) and tracheobronchial lymph node (TBLN) tissue were collected for isolation of mononuclear cells (MNCs). After recall stimulation with A / Swine / OH / 24366 / 2007 or A / California / 04 / 2009 at an MOI of 0.1, the frequency of T cell subsets secreting IFN-γ and IL-17A was measured using intracellular cytokine staining (ICS) flow cytometry as described above, depending on the challenge virus used in each group. Briefly, 5 million cells were seeded in 1 ml of culture medium (RPMI 1640 10% FBS) per well of a 48-well flat-bottom plate and incubated in vitro for 48 hours (hrs) in the presence of recombinant porcine IL-2 and OH / 07 or CA / 09 (0.1 multiplicity of infection - MOI). During the last 6 hours of incubation, the protein transport inhibitor Brefeldin A (GolgiPlug) was added. At the end of incubation, cells were harvested, washed, blocked with 1% normal rabbit serum, and divided into the appropriate number of wells of a 96-well round-bottom plate for surface and intracellular cytokine labeling. Appropriate isotype control antibodies were included as negative controls. For the FACS panel with purified / unlabeled monoclonal antibodies (mAbs), cells were first labeled with purified mAbs and their corresponding secondary antibodies, then blocked with 1% normal mouse serum. Following this step, other cell markers were labeled together as a cocktail. Cells were transferred to a 96-well round-bottom plate, washed twice with 200 μl of FACS buffer per well, and surface labeled with pre-titrated concentrations of fluorochrome-conjugated mAbs against the indicated markers and their corresponding isotype controls in 50 μl of FACS buffer at 40°C for 30 minutes. Cells were then fixed with 1% paraformaldehyde at 4°C for 30 minutes and resuspended in 200 μl of FACS buffer.

[0996] For intracellular labeling, the cells were washed once and permeabilized with 1% saponin at room temperature for 45 minutes. Subsequently, the cells were washed with saponin wash buffer (0.1% saponin) and incubated with a pre-titrated concentration of mAb conjugated to a fluorescent dye for the indicated marker and its corresponding isotype control in a saponin wash solution containing 1% normal rabbit serum at 4°C for 45 minutes in a final volume of 50 μl. The cells were washed once in saponin wash and labeled with the indicated secondary antibody for 45 minutes at 4°C. The cells were washed once and resuspended in 200 μl FACS buffer, transferred to a FACS tube, and analyzed using a BD FACS Aria II flow cytometer through a live cell gate. For each sample, 100,000 events were collected. The data were analyzed using FlowJo software (FlowJoV10, Becton, Dickinson & Company; BD) and plotted using GraphPad Prism (GraphPad Prism 9, CA).

[0997] Use specific immune marker to carry out T helper / memory cell (CD3+CD4+CD8α+β-) and cytotoxic T lymphocyte (CTL) (CD3+CD4+CD8α+β+) immunostaining with the concentration of previous titration and optimization, described specific immune marker comprises the anti-pig CD3 antibody (Southern biotech, AL) of purification or fluorescent dye labeling, CD4α antibody (Southern biotech, AL), CD8α antibody (Southern biotech, AL), CD8β chain antibody (BD Pharmingen, CA), IFN-γ and IL-17A and their corresponding isotype control.Positive and negative colonies, quadrant markers are set, and these are controlled by unstained sample and the sample that is only hatched with isotype control antibody.Initially pass through CD3+ and CD3-gate separation lymphocyte subsets.The lymphocyte frequency of every kind of independent type is expressed as the frequency (percentage) in these cell counts 100,000 cells.

[0998] Example 2

[0999] ORFV Δ121 Generation and characterization of conH1 recombinant viruses

[1000] Phylogenetic analysis based on nucleotide sequences showed the designed conH1 ( Figure 1A Compared with the HA sequences of the recently circulating strains of the H1 subtype from the computer-designed conH1, the genetic distance between conH1 was significantly shortened ( Figure 1B The full-length consensus H1 protein of IAV-S was inserted into locus 121 (ORFV121) of the ORFV genome by homologous recombination ( Figure 1C After deletion and purification, the deleted ORFV121 gene sequence was not detected in the purified recombinant virus ( Figure 1D ), whereas conH1 sequences were detected in the recombinant virus but not in the wild-type ORFV genome ( Figure 1E Western blotting (WB) was used to determine the expression of ORFV Δ121 The presence of conH1 and the efficient processing and cleavage of conH1 HA0 into HA1 and HA2 subunits were detected in conH1-infected cells ( Figure 1F ). ORFV Δ121 The complete genomic sequence of the conH1 recombinant virus confirmed the insertion of the full-length conH1 sequence of IAV-S, the integrity of the ORFV genome, and the complete absence of the virulence determinant ORFV121 (data not shown).

[1001] ORFV Δ121 The replication characteristics of conH1 are similar to those of wild-type (WT) virus (OV IA82) in OFTu cells ( Figure 2A ). In contrast, WT OV-IA82 and ORFV were observed in primary STu cells. Δ121 The significant growth defect of conH1 ( Figure 2B ), indicating minimal or no viral replication in cells of porcine origin, as previously found in other studies.

[1002] Example 3

[1003] Recombinant ORFV Δ121 conH1 expression in vitro

[1004] The expression of ORFV was assessed by immunofluorescence assay (IFA) and flow cytometry. Δ121 The expression of conH1 by recombinant ORFV was assessed during viral infection of OFTu cells by using anti-Flag mAb, IFA and flow cytometry. Δ121 Importantly, these studies demonstrated the expression of conH1 intracellularly, as indicated by strong fluorescence in permeabilized cells, as well as on the surface of infected cells, as shown by non-permeabilized ORFV Δ121 As shown by immunofluorescence staining in conH1 infected cells ( Figure 3A and Figure 3B ). In addition, in the case of continuous passage of ORFV Δ121 IFA assays performed in cells infected with conH1 showed that the recombinant virus stably expressed the inserted full-length conH1 at passages 1, 5, and 10. In conclusion, these results confirm that conH1 is a promising candidate for the treatment of inflammatory bowel disease in vitro. Δ121Robust expression in cells infected with conH1 virus.

[1005] Example 4

[1006] Cross-reactivity between swine H1N1 antiserum and ConH1 in vitro

[1007] The ability of antibodies present in porcine antisera induced by infection with different H1N1 strains to recognize B cell epitopes in conH1 was investigated. Three different antisera from H1N1 strains isolated from 2005 to 2009 representing different IAV-S clades circulating in the United States were used in this assay: anti-A / California / 04 / 2009 (H1N1) belonging to the pandemic clade, anti-A / Swine / IL / 00685 / 2005 (H1N1) from the delta 2 clade, and anti-A / Swine / Kentucky / 02086 / 2008 (H1N1) from the beta clade. Cross-reactivity and antibody binding to conH1 were observed with all three antisera, indicating that the recombinant ORFV Δ121 conH1 expressed by the conH1 virus is recognized by antibodies present in the sera of pigs infected with different H1N1 viruses ( Figure 4A The conH1 amino acid sequence was compared with the HA proteins of these cross-reactive viruses to assess the genetic relationship between these viruses ( Figure 4B and Figure 4C ). Used to construct ORFV Δ121 The conH1 sequence of conH1 has minimal amino acid differences (12.4%) with the HA sequence of virus A / Swine / IL / 00685 / 2005 (H1N1). However, higher cross-reactivity was found using A / California / 04 / 2009 (86.2% pairwise identity) and A / Swine / Kentucky / 02086 / 2008 (84.2% pairwise identity) sera.

[1008] Example 5

[1009] ORFV Δ121 conH1 recombinant virus induces humoral immune response in pigs

[1010] On DPC 0, when whole virus ELISA was performed using challenge virus antigen, IAV-S specific total IgG antibodies were significantly higher in the immunized group ( Figure 5A and Figure 5B The expression of ORFV was also explored by whole virus ELISA against a panel of 10 different viruses at DPC 0. Δ121The breadth of the IAV-S IgG immune response induced by conH1 (Table 2). On DPC0, in the group of animals further challenged with OH / 07, using the whole A / Swine / South Dakota / A02524887 / 2020 virus as an antigen, the immunized group showed a significant increase in the total IgG level in the serum. Similar to OH / 07, the A / Swine / South Dakota / A02524887 / 2020 virus also belongs to the American H1N1 γ branch and is known as the classic swine strain worldwide. It is worth noting that the nucleotide identity between the HA of conH1 used to generate the recombinant virus and the HA of A / Swine / South Dakota / A02524887 / 2020 is 88.1%, and at the amino acid level, the similarity is 89.1% (Table 2). Within the panel of viruses used for the whole virus ELISA, these figures show that the HA of A / Swine / South Dakota / A02524887 / 2020 has the second highest homology to the HA of conH1, only after OH / 07, with 89.2% and 89.8% pairwise identity, respectively (Table 2). On the other hand, in animals challenged with CA / 09, at DPC 0, the vaccinated group showed a significant increase in total IgG levels against A / Swine / South Dakota / A02524887 / 2020 (γ branch), A / Swine / Michigan / A02524810 / 2020 (npdm branch), A / Swine / Texas / A02245632 / 2020 (β branch), A / Swine / Oklahoma / A02245707 / 2020 (β branch), and A / Swine / Oklahoma / A02214419 / 2017 (δ1 branch). Interestingly, these five viruses belong to different H1N1 branches with amino acid homology ranging from 83% to 89% (Table 2). No difference was found between unvaccinated and vaccinated animals using the challenge virus as an antigen at DPC 6. In the group challenged with OH / 07 (γ branch), the vaccinated group maintained a significant increase in total IgG levels using A / Swine / South Dakota / A02524887 / 2020 (γ branch) whole virus as an antigen. In addition, an increase in total IgG antibody levels was detected against A / Swine / Michigan / A02524810 / 2020 (npdm branch), wherein the HA sequence has 84.3% and 85.3% identity to nucleotides and amino acids, respectively (Table 2).In addition, within the group challenged with CA / 09 (npdm clade), the vaccinated group continued to show increased IgG against whole virus antigens of A / Swine / South Dakota / A02524887 / 2020 (γ clade), A / Swine / Michigan / A02524810 / 2020 (npdm clade), A / Swine / Oklahoma / A02245707 / 2020 (β clade), and A / Swine / Oklahoma / A02214419 / 2017 (δ1 clade).

[1011] ORFV Δ121 The conH1 recombinant virus induced cellular immune responses in pigs. The effects of ORFV on peripheral blood mononuclear cells (PBMCs) and monocytes from bronchoalveolar lavage (BAL) and tracheobronchial lymph nodes (TBLN) were evaluated at DPC 6. Δ121 T cell responses induced by conH1 virus immunization. PBMC secreted IFN-γ (IFNγ+) ( Figure 6A and 6E ) and IL-17 (IL17+) T helper / memory cell (CD3+CD4+CD8α+β-) percentages were significantly higher in the vaccinated group ( Figure 6G ). Similarly, ORFV Δ121 The IFNγ+ cytotoxic T lymphocytes (CTLs) in the conH1 immunization group were also enhanced ( Figure 6G Differences in the percentage of IL17+ CTL were only observed between immunized animals and the corresponding sham-immunized / mock-challenged groups ( Figure 6D and Figure 6H ).

[1012] In BAL, the percentage of CD3+CD8α+CD8β+ T cells secreting IL17A and IFN-γ was induced in the vaccinated group at the time of stimulation, compared with the unvaccinated animals in the group challenged with OH / 07 on DPC 6 ( Figure 7A and Figure 7B ), but no difference in IFN-γ secreting cells was found in the group receiving CA / 09 challenge ( Figure 7C and Figure 7D ).

[1013] In contrast, at DPC 6, the levels of IL-17A+ and IFNγ+ secreting T cells CD3+CD8α+CD8β+ in TBLNMNC of pigs vaccinated and challenged with CA / 09 were significantly higher than those in sham-immunized pigs ( Figure 8C and Figure 8D ), while no difference was found in the OH / 07 challenge group ( Figure 8A and Figure 8B ).

[1014] Viral shedding after intranasal IAV-S challenge. ORFV was assessed during challenge with OH / 07 and CA / 09. Δ121 The protective efficacy of conH1 was 10-fold lower in the nasal swabs of the vaccinated group compared to their corresponding sham-immunized group at DPC2 ( Figure 9A and Figure 9B ), indicating early control of viral replication in immunized animals. A significant decrease (approximately 20-fold) in infectious virus and genome copy number was also observed in nasal swabs of the vaccinated group compared with their corresponding sham-immunized group at DPC6 ( Figure 9A and Figure 9B These results indicate that the use of ORFV Δ121 ConH1 vaccination resulted in reduced infectious virus shedding and lower genomic viral loads in nasal secretions after IAV-S challenge with different H1N1 challenge viruses.

[1015] The macroscopic lesions in the lungs were also assessed. Δ121 The lesion score of the conH1 immunization group was significantly lower ( Figure 10A Finally, at DPC6, the genome copies / ml (log 10) and TCID 50 / ml to determine the pseudo-immunized pigs and ORFV Δ121 Viral loads in BAL of conH1-immunized pigs. Although 10-fold lower IAV-S RNA loads were detected in BAL of vaccinated animals, sham-immunized and ORFV Δ121 There were no significant differences in infectious virus titers between the conH1 immunization groups ( Figure 10B In summary, these results indicate that the use of ORFV Δ121 ConH1 vaccination reduced viral shedding following intranasal infection with different strains of IAV-S H1N1 (OH / 07 and CA / 09) and reduced lung lesions following infection with a pandemic virus isolate (CA / 09).

[1016] Example 6

[1017] Materials and methods

[1018] Cells and viruses

[1019] Primary sheep fetal turbinate (OFTu), porcine fetal turbinate (STu) and Madine Derby canine kidney (MDCK) cells were used for virus propagation and cell-based assays. Cells were cultured in minimal essential medium (MEM) supplemented with 10% fetal bovine serum (FBS, heat inactivated), 2 mM L-glutamine and antibiotics (penicillin, streptomycin and gentamicin). Wild-type ORFV (OV strain IA82) was provided by Dr. Daniel Rock of the University of Illinois at Urbana-Champaign (7, 30, 31) and used as a parental virus to produce recombinant ORFV viruses expressing heterologous HA proteins derived from avian (AIV, head) and swine influenza viruses (IAV-S, handle). Both wild-type (wt) and recombinant Orf viruses were propagated in primary OFTu cells.

[1020] Two H1N1 IAV-S strains, A / Swine / OH / 24366 / 2007 (Oh07) (from the gamma clade) and A / California / 04 / 2009 (Ca09) (from the "new epidemic" (npdm) clade) (provided by Dr. Aradhya Gourapura of Ohio State University and obtained from the National Veterinary Services Laboratories (NVSL) (Ames, Iowa), respectively) were used to evaluate the efficacy of recombinant vaccine candidates. IAV-S was amplified in MDCK cells using DMEM supplemented with HEPES buffer and 2 μg / mL of TPCK-treated trypsin. Subsequently, these viruses were used for viral challenge and antigen preparation for whole-virus ELISA.

[1021] For whole virus IgG ELISA, an additional panel of 10 IAV-S isolates was obtained from NVSL to include representatives of all H1 clades. These isolates are listed in Table 5.

[1022] Table 5. 12 IAV-S groups belonging to the H1 subtype used in different assays in this study

[1023] Table 5 (continued)

[1024] Table 5 (continued)

[1025] Table 5 (continued)

[1026] Construction of recombinant expression cassette

[1027] To generate the two recombinant ORFV-SIV viruses evaluated in this study, chimeric-HA (cHA) coding sequences were designed computationally, chemically synthesized, and cloned into the pUC57 plasmid (Genscript®, Piscataway, NJ). Briefly, cHAs were designed based on the stalk domain (nucleotide residues 1033 to 1698) of a contemporary H1N1 IAV-S strain [(A / Swine / Minnesota / A02245569 / 2020 (H1N1) GenBank Accession No. MT372532], and selected to avoid foreign head domains of IAV subtypes present in swine (H1 and H3) and those associated with HPAI (H5 and H7). The head domains of the cHAs used in this study were based on avian influenza A viruses (AIVs) belonging to the H6 subtype [(A / American Green-Winged Teal / Ohio / 18OS2656 / 2018 (H6N1) GenBank Accession No. MN430905.1] or the H8 subtype [(A / Mallard / Ohio / 18OS1248 / 2018 (H8N4) GenBank Accession No. MN431074.1] ( Figure 11B To design chimeric-HA, cH6 / 1, and cH8 / 1, DNA was inserted into the ORFV genome. ORFV121 The restriction endonuclease sites required for the cH6 / 1 locus were added, while HindIII and SalI were added to the N and C termini of cH6 / 1, and SpeI and SalI were added to the N and C termini of cH8 / 1. A Flag tag epitope was added to the 3' end, and p116, the early / late ORFV internal promoter, or the vaccinia virus late I1L promoter was added to the 5' end of the coding sequence of cH6 / 1 and cH8 / 1, respectively. Finally, any potential transcription termination nucleotide sequences, including the TTTTTNT poxvirus transcription termination signal, were removed from the chimeric HA coding sequence by silent nucleotide substitution. The resulting synthetic DNA fragment was subcloned into the poxvirus transfer vector pUC57-ORFV Δ121 loxP-EGFP, generating two recombinant expression cassettes, pUC57-ORFV Δ121 cH6 / 1-loxP-EGFP and pUC57-ORFV Δ121 Correct cloning was verified by restriction enzyme restriction followed by indirect immunofluorescence assay (IFA) of transfected cells using a Flag tag mouse monoclonal antibody (Thermo Fisher Scientific, Waltham, MA).

[1028] Generation of recombinant ORFV Δ121 cH6 / 1 and ORFVΔ121 cH8 / 1-IAV-S

[1029] To generate recombinant ORFV Δ121 cH6 / 1 and ORFV Δ121 cH8 / 1-IAV-S virus was generated by lipofectamine 3000 (Life Technologies, Carlsbad, CA) via the interaction of wt OV IA82 with the recombinant plasmid pUC57-ORFV. Δ121 cH6 / 1-loxP-EGFP and pUC57-ORFV Δ121 Homologous recombination between cH8 / 1-loxP-EGFP inserted the full-length sequences of cH6 / 1 and cH8 / 1 ORFV121 Locus ( Figure 11C and Figure 11D ), as described by Joshi et al. 2021. The presence of chimeric HA and the absence of wild-type virus were confirmed by electrophoresis analysis of PCR amplicons on 1% agarose gels using two pairs of primers designed to PCR amplify the stalk domain of H1 and the internal region of ORFV121 (401 bp), respectively. The primers for the stalk domain of the chimeric HA were Fw-5'-ACTGCGGTACCTATTTAAAAGTTGTTTGGTGAACTTAAATGGG-CCTATTCGGGGCCATTGC-3' (SEQ ID. NO: 17) and Rv-5'-GAGGTCTCGAGTTACTTGTCGTCATCGTCTTTG-TAGTCAATCT-GGTAGATCTTTGTTGAGTCCAGC-3' (SEQ ID. NO: 18) (637 bp), while the primers for ORFV121 (401 bp) were as described previously. PCR was performed using Q5® Hot Start High-Fidelity DNA Polymerase according to the manufacturer's recommendations. For each reaction, a specific annealing temperature (62°C for amplifying the cHA1 handle fragment and 56.5°C for amplifying the ORF121 locus fragment) was used based on the optimal melting temperature (Tm) of the primers used. To verify the presence and integrity of the heterologous gene and ensure the identity and integrity of OV IA82 and the absence of ORFV121, whole-genome sequencing was performed using the MinION Mk1B sequencing platform (Oxford Nanopore Technologies, ONT).

[1030] ORFV Δ121 cH6 / 1 and ORFV Δ121 In vitro characterization of cH8 / 1-IAV-S

[1031] ORFVΔ121 cH6 / 1 or ORFV Δ121 The replication kinetics of the cH8 / 1-IAV-S recombinant virus were assessed by single-step and multistep growth curves in OFTu and STu cells and compared with the parental OV-IA82 as described previously.

[1032] To evaluate heterologous gene expression by recombinant viruses, ORFV Δ121 cH6 / 1 or ORFV Δ121 cH8 / 1-IAV-S infected OFTu cells and evaluated by indirect immunofluorescence assay (IFA), flow cytometry, and Western blotting as described previously.

[1033] The stability of the cH6 / 1 and cH8 / 1 genes inserted into the ORFV121 locus was assessed by evaluating expression by IFA during 10 serial passages of each recombinant virus in OFTu cells, as described previously.

[1034] Vaccination challenge studies in pigs

[1035] Evaluation of ORFV in pigs Δ121 cH6 / 1 or ORFV Δ121 A total of 42 3-week-old pigs (IAV seronegative) were randomly divided into the following four experimental groups: Group 1 - ORFV Δ121 cH6 / 1 (primary) +ORFV Δ121 cH8 / 1-IAV-S (boost) immunization / Oh07 challenge (n = 7); Group 2, ORFVΔ121cH6 / 1 (primary) + ORFV Δ121 cH8 / 1-IAV-S (boost) immunization / Ca09 challenge (n = 7), Group 3, sham immunization / Oh07 challenge (n = 7), and Group 4 sham immunization / Ca09 challenge (n = 7).

[1036] By intramuscular injection of 2 ml containing 10 7.38 Immunization was performed with TCID50 / ml of virus suspension (Groups 1 and 2) or 2 ml of MEM (Groups 3 and 4). Animals were immunized on day 0 (D0) and boosted on day 21 (D21). Two different viruses were selected for challenge: Sw / Oh / 24366 / 2007 (H1N1) (Oh07) and A / California / 04 / 2009 (H1N1) (Ca09). On day 35 (D35), the animals were challenged with 1×10 7 Pigs in Groups 1 and 3 were challenged intranasally with a viral suspension of TCID50 / ml IAV-S Oh07, while pigs in Groups 2 and 4 were challenged intranasally with 1×107 The animals were challenged intranasally with a TCID50 / ml IAV-S Ca09 virus suspension.

[1037] Throughout the study, animals were monitored daily for clinical symptoms of influenza infection. Serum and whole blood samples were collected on days 0, 21, 28, and 35 after immunization (D0, D21, D28, and D35) and on days 3, 5, and 7 after challenge (DPC3, DPC5, and DPC7). Nasal swabs were collected on DPC0, DPC3, DPC5, and DPC7. Animal studies were conducted at Cornell University, following the guidelines and protocols approved by the Institutional Animal Use and Care Committee (IACUC approval number 2019-0041) and in accordance with the provisions of the Animal Welfare Act Amendments.

[1038] Antibody isotype ELISA

[1039] The levels of specific IgG antibodies in serum and IgA antibodies in bronchoalveolar lavage fluid (BAL) were determined by whole-virus ELISA against 12 different IAV-S strains, following a previously described method with some modifications. IgG (Bethyl Laboratories, catalog number: A100-104, TX). Specifically, Immulon 1B ELISA plates (Thermo Fisher Scientific, catalog number: 3355) were coated with concentrated heat-inactivated virus at 500 ng / well instead of 250 ng and incubated with the corresponding goat anti-porcine biotinylated antibody IgG or IgA (Bethyl Laboratories, catalog number: A100-104 or A100-102A, TX, respectively), which was diluted 1:2000 instead of 1:4000 in 5% skim milk Tris-buffered saline (PBST) containing 0.05% Tween 20 detergent.

[1040] Serum samples were evaluated by measuring their optical density (OD) at 450 nm using a microplate reader (SYNERGY LX, BioTek, Winooski, VT, USA). To normalize OD values, each test and control sample was compared to the OD value of an uncoated well. Prior to actual testing, all assays were optimized using serum samples from animals with known serological status.

[1041] Viral load and infectious virus in nasal secretions and tissues

[1042] Viral loads in nasal secretions, bronchoalveolar lavage (BAL), and lungs were determined by real-time reverse transcriptase polymerase chain reaction (rRT-PCR). Lung lysates for RNA extraction were prepared from 2 g of lung tissue homogenized in 20 mL of MEM from a single pig. Approximately 200 μl of tissue homogenate supernatant was subjected to nucleic acid extraction using the MagMAX Core extraction kit (Thermo Fisher, Waltham, MA, USA) and the Kingfisher Flex automated extraction platform as described previously. Path-ID TM RNA from influenza A virus was detected using real-time RT-PCR targeting the conserved matrix (M) gene (Integrated DNA Technologies; Coralville, IA, USA) using a Multiplex One-Step RT-PCR Kit (Thermo Fisher, Waltham, MA, USA). The probe and forward and reverse primers used were 5'-FAM-TCA GGC CCC CTC AAA GCCGA-BHQ1 -3' (SEQ ID. NO: 19), 5'- AGA TGA GTC TTC TAA CCG AGG TCG -3' (SEQ ID. NO: 20), and 5'- TGC AAA AAC ATC TTC AAG TCT CTG -3' (SEQ ID. NO: 21), respectively. The cycling conditions for amplifying the M gene target were: reverse transcription at 48°C for 10 min, polymerase activation at 95°C for 10 min, 45 cycles of denaturation at 95°C for 15 s and annealing and extension at 60°C for 1 min, using a CFX96 Touch Real-Time PCR Detection System (BioRad). -1 to 10 -8 A standard curve was established using 10-fold serial dilutions of PCR products. As previously described, the genomic copies / mL (log10) for each sample was derived from the CT obtained by the standard curve established using CFX Maestro software (Bio-Rad). Appropriate positive and negative controls were added to each extract and PCR plate and run alongside the test samples.

[1043] In addition, as described in previous studies, the infectious virus in nasal swabs and tissue samples was assessed in MDCK cells and expressed as TCID50 / mL. Mouse monoclonal antibodies (mAbs) for IFA virus titration were provided by Dr. Eric Nelson and Dr. Steve Lawson of SDSU (IAV-NP Hb-65462 mAb; 1:500). This antibody targets the conserved nucleoprotein (NP) of influenza A virus. The appearance of fluorescent foci indicates virus-positive wells.

[1044] Statistical analysis

[1045] GraphPad Prism software v9.0 (GraphPad Software, San Diego, CA, USA) was used for all statistical analyses. A Shapiro-Wilk test was performed to verify data normality, followed by a t-test or unpaired t-test to compare the mean values ​​between normal and non-normal data groups, respectively. Paired comparisons were performed using Tukey's multiple comparison post hoc test. P-values ​​less than 0.05 were considered significant. Flow cytometry data were acquired using an Attune NxT flow cytometer and analyzed using FlowJo software (FlowJo V10, Becton, Dickinson & Company; BD, Franklin Lakes, NJ, USA).

[1046] Example 7

[1047] In vitro expression of heterologous proteins by ORFVΔ121cH6 / 1 and ORFVΔ121cH8 / 1 viruses

[1048] The expression of cH6 / 1 and cH8 / 1 by each recombinant virus was verified by IFA, WB and flow cytometry using anti-FLAG mAb. Figure 12A ) and ORFVΔ121cH8 / 1 ( Figure 12B ) infected cells, red fluorescence was observed inside the cells (permeabilized) and on the cell surface (non-permeabilized). Notably, the level of chimeric HA protein (about 70 kDa) detected by the flag epitope was significantly higher in the recombinant ORFVΔ121cH6 / 1 ( Figure 12C ) and ORFVΔ121cH8 / 1 ( Figure 12D ) increased to 72 hpi in sheep cells infected at 10 MOI. Flow cytometry data reinforced the results from IFA and WB assays ( Figure 12E and Figure 12F ).

[1049] In addition, the expression of cH6 / 1 and cH8 / 1 was assessed by flow cytometry at 72 hpi. The recombinant ORFV was observed intracellularly in approximately 56% and 47% of the cells and on the plasma membrane in 31% and 25% of the infected cells, respectively. Δ121 cH6 / 1 and ORFV Δ 121 cH8 / 1 virus expresses cH6 / 1 and cH8 / 1.

[1050] PCR amplification ( Figure 13A ) and whole genome sequencing confirmed the presence of Δ121 cH6 / 1 and ORFVΔ121 The integrity of the chimeric HA sequence inserted into the cH8 / 1 virus was assessed by multi-step and single-step growth curves in OFTu and STu cells. Δ 121 cH6 / 1 and ORFV Δ121 Replication kinetics of cH8 / 1-IAV-S ( Figure 13B As expected, both viruses replicated to high titers in OFTu cells, whereas a clear impairment of replication was observed in primary porcine cells.

[1051] Example 8

[1052] Pig humoral immune response

[1053] In the presence of two challenge viruses, ORFV Δ121 cH6 / 1 as primary immunization and ORFV Δ121 In animals immunized sequentially with cH8 / 1-IAV-S as a booster (OVCH6 / 1-CH8 / 1), IAV-S-specific total IgG antibodies were detected as early as 3 weeks after primary vaccination and increased significantly after the D21 booster vaccination, remaining elevated until D42 (7 dpc) ( Figure 14A Similar results were detected using Oh07 or Ca09 purified viruses as antigens in ELISA ( Figure 14B ). After Oh07 and CA09 virus challenge infection, memory responses were observed in both groups. It is worth noting that IgG levels reached their peak on D42 (or DPC7), indicating that IgG memory B cells generated after immunization were activated during experimental challenge infection.

[1054] The breadth of IAV-S-specific IgG immune responses induced by vaccination was also assessed against a panel of 10 different viruses on D35 (DPC0) (Table 5). Δ121 cH6 / 1 and ORFV Δ121 Animals immunized with cH8 / 1 virus for the first and second time showed significantly higher IgG responses ( Figure 15A and Figure 15B These results demonstrate that the antibodies induced by IM immunization of the chimeric virus in pigs have broad binding capacity.

[1055] Example 9

[1056] Pulmonary mucosal immunity

[1057] Mucosal antibody responses in vaccinated pigs compared to mock-immunized pigs were assessed by measuring IgA antibody levels in BAL after euthanasia on D42 against the same panel of different viruses used in the previous ELISA assay.

[1058] Compared with sham-immunized pigs, immunized animals in the OH07 challenge group showed significantly higher IgA levels against all 12 IAV-S viruses ( Figure 16A On the other hand, the vaccinated pigs challenged with CA09 virus showed significantly higher IgA responses to all the different viruses except SD / 18, Missouri / 20 and Oh / 07 which belonged to the gamma-2-beta-like, npdm and gamma clades, respectively ( Figure 16B These results indicate that ORFV Δ121 cH6 / 1 and ORFV Δ121 IM immunization with cH8 / 1 virus elicits a cross-reactive mucosal immune response.

[1059] Example 10

[1060] Nasal swab viral shedding

[1061] After intranasal challenge with Oh07 or Ca09 virus, the presence of viral RNA and infectious virus in nasal secretions was assessed. Animals immunized with OV-cH6 / 1-cH8 / 1 and subsequently challenged with Oh07 showed approximately a 10-fold reduction in infectious virus shedding through nasal secretions on day 1 of the immunization, and this difference reached 20-fold on day 5 of the immunization compared to the corresponding sham-immunized group ( Figure 17A ). Similar results were observed for viral RNA loads as determined by RT-qPCR. Peak viral shedding in OV-cH6 / 1-cH8 / 1 + Oh07 challenged pigs was at DPC3, with no significant increase in infectious virus and genome copy number from DPC1 to DPC3. In summary, these results indicate early control of Oh07 replication and early viral clearance in immunized animals. In contrast, no differences were found in the Ca09 challenged group, except for a slight decrease in viral RNA in the vaccinated group at DPC5 ( Figure 17B For both challenges, no infectious virus or IAV-S genome copies were found in any group at DPC 7. These results indicate that vaccination with OV-cH6 / 1-cH8 / 1 resulted in a reduction in viral shedding in nasal secretions following Oh07 infection, but not Ca09.

[1062] Example 11

[1063] Local lesions and viral load

[1064] Macroscopic lesions present in the lungs were scored based on the method developed by Madec and Kobisch. Tissue consolidation, represented by dark plaque areas, was more pronounced in sham-immunized animals ( 19A to 19DIn the OV-cH6 / 1-cH8 / 1 immunization group, the lesion score was significantly lower regardless of the challenge virus ( Figure 18A and Figure 18B In animals sham-immunized + Oh07-challenged, lesion scores were approximately 10-fold higher, which may be due to the higher pathogenicity of this virus strain in pigs. In addition, on D42 (DPC7), viral loads in lung lysates and BAL were determined and expressed as genome copies / mL (log 10). In vaccinated animals, viral loads in both lung and BAL were 10-fold lower ( Figure 18B In summary, these results indicate that the use of ORFV Δ121 cH6 / 1-ORFV Δ 121 cH8 / 1-IAV-S vaccination reduced viral load and lung lesions.

Claims

1. An amino acid sequence comprising a consensus H1 hemagglutinin polypeptide.

2. The amino acid sequence of claim 1, wherein the consensus H1 hemagglutinin polypeptide comprises SEQ ID NO:

1.

3. The amino acid sequence of claim 1, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO:

1.

4. The amino acid sequence of claim 1, wherein the consensus H1 hemagglutinin polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO:

1.

5. A nucleotide sequence comprising a nucleic acid sequence encoding a consensus H1 hemagglutinin polypeptide.

6. The nucleotide sequence of claim 5, wherein the nucleic acid sequence comprises SEQ ID NO:

2.

7. The nucleotide sequence of claim 5, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO:

2.

8. The nucleotide sequence of claim 5, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO:

2.

9. A composition comprising a consensus H1 hemagglutinin polypeptide and a carrier.

10. The composition of claim 9, wherein the consensus H1 hemagglutinin polypeptide comprises an amino acid sequence comprising SEQ ID NO:

1.

11. The composition of claim 9, wherein the vector is a viral vector.

12. The composition of claim 11, wherein the viral vector is an ORFV vector.

13. The composition of claim 9, wherein the vector is a nucleic acid vector.

14. A pharmaceutical composition comprising: i) a consensus H1 hemagglutinin polypeptide, and ii) one or more pharmaceutically acceptable carriers.

15. The pharmaceutical composition of claim 14, wherein the consensus H1 hemagglutinin polypeptide is comprised in the composition of any one of claims 9 to 13.

16. The pharmaceutical composition of claim 14, wherein the consensus H1 hemagglutinin polypeptide comprises an amino acid sequence comprising SEQ ID NO:

1.

17. The pharmaceutical composition of claim 14, wherein the pharmaceutical composition is formulated as a vaccine.

18. A method of inducing an immune response against influenza A virus (IAV) in an animal, the method comprising the step of administering to the animal a therapeutically effective amount of the pharmaceutical composition of any one of claims 14 to 17.

19. The method of claim 18, wherein the immune response comprises a cytotoxic T lymphocyte (CTL) response.

20. The method of claim 18, wherein the immune response comprises an IgG response.

21. The method of claim 18, wherein the immune response comprises a T cell response.

22. The method of claim 18, wherein the immune response comprises a T helper / memory cell response.

23. The method of claim 18, wherein the immune response comprises an IFN-γ response.

24. The method of claim 18, wherein the administration is prophylactic administration to the animal.

25. The method of claim 18, wherein the animal is porcine.

26. The method of claim 18, wherein the method comprises reducing the IAV viral load in the animal.

27. The method of claim 18, wherein the method comprises reducing IAV viral shedding in the animal.

28. A method for preventing influenza A virus (IAV) infection in an animal, the method comprising the step of administering to the animal a therapeutically effective amount of the pharmaceutical composition of any one of claims 14 to 17.

29. The method of claim 28, wherein the administration is prophylactic administration to the animal.

30. The method of claim 28, wherein the animal is porcine.

31. The method of claim 28, wherein the method comprises reducing the IAV viral load in the animal.

32. The method of claim 28, wherein the method comprises reducing IAV viral shedding in the animal.

33. An amino acid sequence comprising a consensus H3 hemagglutinin polypeptide.

34. The amino acid sequence of claim 33, wherein the consensus H3 hemagglutinin polypeptide comprises SEQ ID NO:

3.

35. The amino acid sequence of claim 33, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO:

3.

36. The amino acid sequence of claim 33, wherein the consensus H3 hemagglutinin polypeptide comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO:

3.

37. A nucleotide sequence comprising a nucleic acid sequence encoding a consensus H3 hemagglutinin polypeptide.

38. The nucleotide sequence of claim 37, wherein the nucleic acid sequence comprises SEQ ID NO:

4.

39. The nucleotide sequence of claim 37, wherein the nucleic acid sequence comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO:

4.

40. The nucleotide sequence of claim 37, wherein the nucleic acid sequence comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO:

4.

41. A composition comprising a consensus H3 hemagglutinin polypeptide and a carrier.

42. The composition of claim 41, wherein the consensus H3 hemagglutinin polypeptide comprises an amino acid sequence comprising SEQ ID NO:

3.

43. The composition of claim 41, wherein the vector is a viral vector.

44. The composition of claim 43, wherein the viral vector is an ORFV vector.

45. The composition of claim 41, wherein the vector is a nucleic acid vector.

46. ​​A pharmaceutical composition comprising: i) a consensus H3 hemagglutinin polypeptide, and ii) one or more pharmaceutically acceptable carriers.

47. The pharmaceutical composition of claim 46, wherein the consensus H3 hemagglutinin polypeptide is comprised in the composition of any one of claims 41 to 45.

48. The pharmaceutical composition of claim 46, wherein the consensus H3 hemagglutinin polypeptide comprises an amino acid sequence comprising SEQ ID NO:

3.

49. The pharmaceutical composition of claim 46, wherein the pharmaceutical composition is formulated as a vaccine.

50. A method of inducing an immune response against influenza A virus (IAV) in an animal, the method comprising the step of administering to the animal a therapeutically effective amount of the pharmaceutical composition of any one of claims 46 to 49.

51. The method of claim 50, wherein the immune response comprises a cytotoxic T lymphocyte (CTL) response.

52. The method of claim 50, wherein the immune response comprises an IgG response.

53. The method of claim 50, wherein the immune response comprises a T cell response.

54. The method of claim 50, wherein the immune response comprises a T helper / memory cell response.

55. The method of claim 50, wherein the immune response comprises an IFN-γ response.

56. The method of claim 50, wherein the administration is prophylactic administration to the animal.

57. The method of claim 50, wherein the animal is porcine.

58. The method of claim 50, wherein the method comprises reducing the IAV viral load in the animal.

59. The method of claim 50, wherein the method comprises reducing IAV viral shedding in the animal.

60. A method of preventing influenza A virus (IAV) infection in an animal, said method comprising the step of administering to said animal a therapeutically effective amount of the pharmaceutical composition of any one of claims 46 to 49.

61. The method of claim 60, wherein the administration is prophylactic administration to the animal.

62. The method of claim 60, wherein the animal is porcine.

63. The method of claim 60, wherein the method comprises reducing the IAV viral load in the animal.

64. The method of claim 60, wherein the method comprises reducing IAV viral shedding in the animal.

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