Combination vaccines

Abstract

This disclosure relates to a composition comprising multiple target antigens for use in a vaccine formulation. The composition comprises (a) a polynucleotide (such as RNA) containing a nucleotide sequence encoding an antigen, and (b) an antigen polypeptide. The composition can be used to induce an immune response in a target and / or to treat or prevent diseases such as respiratory viral infections.

Description

Technical Field

[0001] Related Application Data This application claims priority from U.S. Patent Application No. 63 / 501,222, filed May 10, 2023, entitled "Combination vaccine", the entire contents of which are incorporated herein by reference.

[0002] Sequence Listing This application is filed with a sequence listing in electronic form. The entire contents of the sequence listing are incorporated herein by reference.

[0003] The present disclosure relates to a composition comprising a plurality of antigens of interest for use in a vaccine formulation.

Background Art

[0004] Respiratory virus infections are a major threat to human health. Infections such as those caused by influenza virus and severe acute respiratory syndrome coronavirus (SARS-CoV) are known to have caused worldwide pandemics and led to the deaths of millions of people around the world. More recently, SAR-CoV-2 has been the cause of the ongoing worldwide pandemic of severe infectious coronavirus disease 2019 (COVID-19). Furthermore, respiratory syncytial virus (RSV) is the most common single cause of hospitalization due to respiratory disease in infants, and reinfection remains common throughout later life. Several vaccines are available for viral infections such as influenza, SARS-CoV-2, and RSV, such as the mRNA vaccines developed to treat or prevent SARS-CoV-2 infection, but further improvements can be made to increase their effectiveness and / or improve treatment strategies.

[0005] Currently, egg-based manufacturing processes are the most common method of vaccine production. This process requires considerable time to optimize viral replication within the eggs and necessitates resources (i.e., eggs) to produce sufficient quantities of vaccine, especially during a pandemic. Furthermore, given the long development time required, vaccine strain selection takes place before the vaccine becomes available, making it difficult to adapt to changes in the virus. Vaccines are also produced using cell-based manufacturing processes involving cultured mammalian cells (e.g., Madin Darby canine kidney, or MDCK cells) instead of eggs, and virus-based platforms involving recombinant viruses (e.g., baculoviruses encoding influenza antigens) are also being utilized.

[0006] There is still a need for the development of specific and efficient viral vaccines that can be produced more rapidly and with broader utility than current egg-based technologies, for the treatment or prevention of respiratory viral infections such as influenza, RSV, and SARS-CoV-2. Nucleic acid-based vaccines offer clear advantages over current egg-based manufacturing platforms, but several challenges remain. For example, due to the inherently unstable nature of mRNA, most RNA-based vaccines consequently have a limited ability to deliver antigens at the doses and durations required to produce a robust and durable immune response.

[0007] Therefore, it will be clear to those skilled in the art that there is a need in the art for compositions having broader utility and / or improved efficacy that are suitable for use as vaccines. [Overview of the project]

[0008] This disclosure is based on the inventors' findings that combining RNA-based antigens with protein-based antigens in immunogenic compositions such as vaccines can enhance the target immune response to the immunogenic composition.

[0009] The inventors further found that co-administration of RNA vaccines, such as auto-amplified mRNA (sa-mRNA) vaccines and adjuvanted protein-based vaccines, resulted in enhanced neutralizing titers for the antigens of both vaccines and affected the subclass of antigen-specific IgG induced by recombinant protein vaccines. For example, the inventors found that the antigen-specific IgG1:IgG2a ratio induced by adjuvanted protein vaccines shifted significantly toward a balanced IgG1:IgG2a response when co-administered with RNA vaccines, such as auto-amplified mRNA (sa-mRNA) vaccines. These improvements were observed, for example, when RNA vaccines and protein vaccines were co-administered distally to each other.

[0010] The inventors have further found that co-administration of RNA vaccines and protein vaccines locally provides similar benefits to co-formulated RNA vaccines and protein vaccines compared to co-administration of RNA vaccines and protein vaccines distal to each other.

[0011] Accordingly, the present inventors' findings provide a basis for an immunogenic composition comprising (i) RNA containing a nucleotide sequence encoding one or more target antigens selected from the group consisting of respiratory syncytial virus (RSV), influenza virus, and / or severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (ii) a polypeptide antigen selected from the group consisting of RSV, influenza virus, and / or SARS-CoV-2. Furthermore, the present inventors' findings provide a basis for a method of treating, preventing, or delaying the progression of a disease or disorder (e.g., RSV, influenza, SARS-CoV-2 infection, COVID-19, or ARDS) in a subject by administering the immunogenic composition of the present disclosure.

[0012] Therefore, this disclosure is, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen selected from the group consisting of respiratory syncytial virus (RSV), influenza, and severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (ii) To provide an immunogenic composition comprising an antigen polypeptide selected from the group consisting of RSV, influenza, and SARS-CoV-2.

[0013] When discussing antigens selected from the group consisting of respiratory syncytial virus (RSV), influenza, and severe acute respiratory syndrome coronavirus (SARS-CoV-2), those skilled in the art will recognize that the antigens are derived from the listed viruses.

[0014] For example, immunogenic compositions, a) Polynucleotides comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0015] In another example, immunogenic compositions, a) Polynucleotides comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0016] In another example, immunogenic compositions, a) Polynucleotides comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) It contains antigen polypeptides from influenza.

[0017] In another example, the immunogenic composition a) is a polynucleotide containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) contains antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0018] In another example, the immunogenic composition a) is a polynucleotide containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) contains antigen polypeptides from respiratory syncytial virus (RSV).

[0019] In another example, the immunogenic composition a) is a polynucleotide containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) contains antigen polypeptides from influenza.

[0020] In another example, the immunogenic composition a) is a polynucleotide containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) contains antigen polypeptides from respiratory syncytial virus (RSV).

[0021] In another example, the immunogenic composition a) A polynucleotide comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) comprises an antigen polypeptide from influenza.

[0022] In another example, the immunogenic composition is a) A polynucleotide comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) comprises an antigen polypeptide from SARS-CoV-2.

[0023] In one example, the immunogenic composition is (i) RNA comprising a nucleotide sequence encoding an antigen selected from the group consisting of respiratory syncytial virus (RSV), influenza virus, and / or severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (ii) comprises an antigen polypeptide selected from the group consisting of RSV, influenza virus, and / or SARS-CoV-2.

[0024] In one example, the immunogenic composition is a) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) comprises an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0025] In another example, the immunogenic composition is a) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0026] In another example, immunogenic compositions, a) RNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0027] In another example, immunogenic compositions, a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0028] In another example, immunogenic compositions, a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0029] In another example, immunogenic compositions, a) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0030] In another example, immunogenic compositions, a) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0031] In another example, immunogenic compositions, a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0032] In another example, immunogenic compositions, a) RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from SARS-CoV-2.

[0033] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen selected from the group consisting of respiratory syncytial virus (RSV), influenza virus, and / or severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (ii) comprising an antigen polypeptide selected from the group consisting of RSV, influenza virus, and / or SARS-CoV-2.

[0034] For example, immunogenic compositions, a) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0035] In another example, immunogenic compositions, a) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0036] In another example, immunogenic compositions, a) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0037] In another example, immunogenic compositions, a) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0038] In another example, immunogenic compositions, a) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0039] In another example, immunogenic compositions, a) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0040] In another example, immunogenic compositions, a) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from RSV

[0041] In another example, immunogenic compositions, a) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0042] In another example, immunogenic compositions, a) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from SARS-CoV-2.

[0043] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen selected from the group consisting of respiratory syncytial virus (RSV), influenza virus, and / or severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (ii) comprising an antigen polypeptide selected from the group consisting of RSV, influenza virus, and / or SARS-CoV-2.

[0044] For example, immunogenic compositions, a) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0045] In another example, immunogenic compositions, a) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0046] In another example, immunogenic compositions, a) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0047] In another example, immunogenic compositions, a) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0048] In another example, immunogenic compositions, a) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0049] In another example, immunogenic compositions, a) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0050] In another example, immunogenic compositions, a) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from RSV

[0051] In another example, immunogenic compositions, a) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0052] In another example, immunogenic compositions, a) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from SARS-CoV-2.

[0053] In one example, the immunogenic composition of this disclosure comprises a plurality of polynucleotides, each polynucleotide encoding a different polypeptide sequence. In another example, the immunogenic composition of this disclosure comprises a plurality of RNAs, each RNA encoding a different polypeptide sequence. In yet another example, the immunogenic composition of this disclosure comprises a plurality of cRNAs, each cRNA encoding a different polypeptide sequence. In yet another example, the immunogenic composition of this disclosure comprises a plurality of self-replicating RNAs, each self-replicating RNA encoding a different polypeptide sequence.

[0054] Therefore, in one example, an immunogenic composition is (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0055] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Polynucleotides containing nucleotide sequences encoding antigens from respiratory syncytial virus (RSV), (ii) Contains a polynucleotide that includes a nucleotide sequence encoding an antigen from influenza.

[0056] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Polynucleotides containing nucleotide sequences encoding antigens from influenza, and (ii) Contains a polynucleotide comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV).

[0057] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0058] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Polynucleotides containing nucleotide sequences encoding antigens from severe acute respiratory syndrome coronavirus (SARS-CoV-2), (ii) Contains a polynucleotide that includes a nucleotide sequence encoding an antigen from influenza.

[0059] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Polynucleotides containing nucleotide sequences encoding antigens from influenza, and (ii) Contains polynucleotides comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0060] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0061] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Polynucleotides containing nucleotide sequences encoding antigens from severe acute respiratory syndrome coronavirus (SARS-CoV-2), (ii) Contains a polynucleotide comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV).

[0062] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Polynucleotides containing nucleotide sequences encoding antigens from respiratory syncytial virus (RSV), and (ii) Contains polynucleotides comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0063] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0064] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0065] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0066] For example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0067] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0068] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0069] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0070] For example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0071] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0072] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0073] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0074] For example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0075] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0076] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0077] In another example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Polynucleotides comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0078] For example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0079] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) RNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), (ii) Contains RNA containing a nucleotide sequence encoding an antigen from influenza.

[0080] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) RNA containing a nucleotide sequence encoding an antigen from influenza, and (ii) RNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV).

[0081] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0082] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) RNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), (ii) Contains RNA containing a nucleotide sequence encoding an antigen from influenza.

[0083] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) RNA containing a nucleotide sequence encoding an antigen from influenza, and (ii) Contains RNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0084] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0085] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) RNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), (ii) RNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV).

[0086] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) RNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and (ii) Contains RNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0087] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0088] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0089] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0090] For example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0091] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0092] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0093] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0094] For example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0095] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0096] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0097] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0098] For example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0099] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0100] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0101] In another example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0102] For example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0103] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) cRNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), (ii) Contains cRNA containing a nucleotide sequence encoding an antigen from influenza.

[0104] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) cRNA containing a nucleotide sequence encoding an antigen from influenza, and (ii) Containing cRNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV).

[0105] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0106] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) cRNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), (ii) Contains cRNA containing a nucleotide sequence encoding an antigen from influenza.

[0107] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) cRNA containing a nucleotide sequence encoding an antigen from influenza, and (ii) Contains cRNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0108] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0109] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) cRNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), (ii) Containing cRNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV).

[0110] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) cRNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and (ii) Contains cRNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0111] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0112] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0113] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0114] For example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0115] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0116] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0117] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0118] For example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0119] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0120] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0121] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0122] For example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0123] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0124] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0125] In another example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0126] For example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0127] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Self-replicating RNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), (ii) Contains self-replicating RNA containing a nucleotide sequence encoding an antigen from influenza.

[0128] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Self-replicating RNA containing a nucleotide sequence encoding an antigen from influenza, and (ii) Contains self-replicating RNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV).

[0129] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0130] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Self-replicating RNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), (ii) Contains self-replicating RNA containing a nucleotide sequence encoding an antigen from influenza.

[0131] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Self-replicating RNA containing a nucleotide sequence encoding an antigen from influenza, and (ii) Contains self-replicating RNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0132] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0133] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Self-replicating RNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), (ii) Contains self-replicating RNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV).

[0134] In one example, the immunogenic composition is arranged in the order from 5' to 3'. (i) Self-replicating RNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and (ii) Contains self-replicating RNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0135] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0136] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0137] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0138] For example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0139] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0140] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0141] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0142] For example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0143] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0144] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0145] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0146] For example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0147] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0148] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0149] In another example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0150] In one example, the immunogenic composition comprises multiple multi-cistronic self-replicating RNAs, each encoding a polypeptide of the target antigen. In another example, the immunogenic composition of this disclosure comprises multiple self-replicating mono-cistronic RNAs, each encoding a different polypeptide sequence. For example, one self-replicating RNA encodes a polypeptide of the target antigen, and different self-replicating RNAs encode different polypeptides of the target antigen.

[0151] In one example of this disclosure, the immunogenic composition comprises multiple polypeptide antigens, each polypeptide being a different antigen of interest. In another embodiment, the different polypeptides of the target antigen may be from the same virus. For example, the different polypeptides may be from respiratory syncytial virus (RSV), severe acute respiratory syndrome coronavirus (SARS-CoV-2), or influenza.

[0152] For example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0153] For example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0154] For example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (iii) Contains an antigen polypeptide derived from influenza.

[0155] For example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from respiratory syncytial virus (RSV), and (iii) Contains further polypeptides from respiratory syncytial virus (RSV).

[0156] For example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Contains further antigenic polypeptides from influenza.

[0157] For example, immunogenic compositions, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (iii) Contains further antigenic polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0158] For example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0159] For example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0160] For example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (iii) Contains an antigen polypeptide derived from influenza.

[0161] For example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from respiratory syncytial virus (RSV), and (iii) Contains further polypeptides from respiratory syncytial virus (RSV).

[0162] For example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Contains further antigenic polypeptides from influenza.

[0163] For example, immunogenic compositions, (i) RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (iii) Contains further antigenic polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0164] For example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0165] For example, immunogenic compositions, (i) cRNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and (iii) It contains antigen polypeptides from respiratory syncytial virus (RSV).

[0166] In one example, the immunogenic composition is (i) cRNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and (iii) It contains antigen polypeptides from influenza.

[0167] In one example, the immunogenic composition is (i) cRNA containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from respiratory syncytial virus (RSV), and (iii) It contains further polypeptides from respiratory syncytial virus (RSV).

[0168] In one example, the immunogenic composition is (i) cRNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from influenza, and (iii) It contains further antigen polypeptides from influenza.

[0169] In one example, the immunogenic composition is (i) cRNA containing a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (iii) Contains further antigenic polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0170] For example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0171] For example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0172] For example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (iii) Contains an antigen polypeptide derived from influenza.

[0173] For example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from respiratory syncytial virus (RSV), and (iii) Contains further polypeptides from respiratory syncytial virus (RSV).

[0174] For example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Contains further antigenic polypeptides from influenza.

[0175] For example, immunogenic compositions, (i) Self-replicating RNA comprising a nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (iii) Contains further antigenic polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0176] For example, the compositions described herein further include adjuvants.

[0177] In one example of this disclosure, the protein antigen is formulated using an adjuvant.

[0178] In one example, the adjuvant is selected from the group consisting of Freund's adjuvant, incomplete Freund's adjuvant, aluminum phosphate, aluminum hydroxide, GMCSP, BCG, MDP compounds such as thur-MDP and nor-MDP, CGP (MTP-PE), lipid A, monophosphoryl lipid A (MPL), RIBI, MPL, trehalose dimycolate (TDM), Novasomes®, QS21, Quil A (and their derivatives and components), calcium phosphate, calcium hydroxide, zinc hydroxide, MHC antigens, PolyI:C, MF59, glycolipid analogs, octadecyl esters of amino acids, muramyl dipeptide, polyphosphazenes, lipoproteins, ISCOM matrix, DC-Chol, ODA, cytokines, and other adjuvants and derivatives thereof. In another example, the adjuvant is MF59.

[0179] In one example, the effect of the immunogenic composition described herein on the treatment, prevention, or delay of the progression of a disease or disorder is enhanced in the presence of an adjuvant such as MF59 compared to the effect of the immunogenic composition described herein that does not contain an adjuvant.

[0180] In one example, the effect of the immunogenic composition described herein on the treatment, prevention, or delay of the progression of SARS-CoV-2 infection, COVID-19, influenza, and / or respiratory syncytial virus (RSV) is enhanced in the presence of an adjuvant such as MF59 compared to the effect of the immunogenic composition described herein that does not contain an adjuvant.

[0181] In one example, the effect of the immunogenic composition described herein on the treatment, prevention, or delay of the progression of pneumonia, sepsis, or acute respiratory distress syndrome in a subject with COVID-19 is enhanced in the presence of an adjuvant such as MF59 compared to the effect of the immunogenic composition described herein that does not contain an adjuvant.

[0182] For example, the effects of the immunogenic compositions described herein on the treatment, prevention, or delay of the progression of influenza infection are enhanced in the presence of an adjuvant such as MF59 compared to the effects of the immunogenic compositions described herein without an adjuvant.

[0183] For example, the effects of the immunogenic compositions described herein on the treatment, prevention, or delay of the progression of RSV infection are enhanced in the presence of an adjuvant such as MF59 compared to the effects of the immunogenic compositions described herein without an adjuvant.

[0184] For example, the effect of the immunogenic compositions described herein on inducing an immune response in a subject is enhanced in the presence of an adjuvant such as MF59 compared to the effect of the immunogenic compositions described herein without an adjuvant.

[0185] For example, the presence of an adjuvant such as MF59 in an immunogenic composition described herein increases the induction of an immune response by the composition by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 300%, at least about 400%, at least about 500%, or more, compared to the effect of an immunogenic composition described herein that does not contain an adjuvant such as MF59.

[0186] In one example, the adjuvant is administered simultaneously with the immunogenic composition of the present disclosure. In another example, the adjuvant is administered immediately before, prior to, or following the administration of the immunogenic composition of the present disclosure.

[0187] For example, the compositions described herein, (i) One or more antigens, (ii) One or more immunostimulants, (iii) One or more chemiactive substances, and / or (iv) Further comprising an additional polypeptide selected from the group consisting of one or more targeted molecules.

[0188] In one example, a nucleotide sequence is operably linked to a regulatory element selected from a group consisting of a promoter, optionally a subgenome (SG) promoter, an internal ribosome entry site (IRES), and a Kozak consensus sequence, or a combination thereof. In one example, the regulatory element is an SG promoter.

[0189] For example, the polynucleotide of the immunogenic composition includes a nucleotide sequence encoding an antigen operably linked to an SG promoter, as well as a regulatory element selected from IRES and / or Kozak consensus sequences.

[0190] In another example, the RNA of the immunogenic composition includes an SG promoter, as well as a nucleotide sequence encoding an antigen operably ligated to a regulatory element selected from IRES and / or Kozak consensus sequences.

[0191] In another example, the cRNA of the immunogenic composition includes an SG promoter, as well as a nucleotide sequence encoding an antigen operably linked to a regulatory element selected from IRES and / or Kozak consensus sequences.

[0192] In another example, the self-replicating RNA of the immunogenic composition includes an SG promoter, as well as a nucleotide sequence encoding an antigen operably linked to a regulatory element selected from IRES and / or Kozak consensus sequences.

[0193] Therefore, this disclosure is, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and b) a second nucleotide sequence encoding an antigen from influenza operably linked to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0194] In one example, the polynucleotide comprises, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and b) a second nucleotide sequence encoding an antigen from influenza, operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0195] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0196] In another example, this disclosure is: (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from influenza operably linked to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from respiratory syncytial virus (RSV).

[0197] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from influenza, operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0198] In one example, the polynucleotide comprises, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0199] In another example, this disclosure is: (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide derived from influenza.

[0200] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0201] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0202] Therefore, this disclosure also, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and (b) a second nucleotide sequence encoding an antigen from influenza operably linked to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) We also provide immunogenic compositions containing antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0203] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and b) a second nucleotide sequence encoding an antigen from influenza, operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0204] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0205] In another example, this disclosure is: (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from respiratory syncytial virus (RSV).

[0206] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from influenza, operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0207] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0208] In another example, this disclosure is: (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide derived from influenza.

[0209] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0210] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0211] Therefore, this disclosure also, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and b) a second nucleotide sequence encoding an antigen from influenza operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) We also provide immunogenic compositions containing antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0212] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and b) a second nucleotide sequence encoding an antigen from influenza, operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0213] In one example, the cRNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0214] In another example, this disclosure is: (i) a cRNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from respiratory syncytial virus (RSV).

[0215] In one example, the cRNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from influenza, operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0216] In one example, the cRNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0217] In another example, this disclosure is: (i) a cRNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide derived from influenza.

[0218] In one example, the cRNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0219] In one example, the cRNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0220] Therefore, this disclosure also, (i) Self-replicating RNA comprising (a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and (b) a second nucleotide sequence encoding an antigen from influenza operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) We also provide immunogenic compositions containing antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0221] In one example, the self-replicating RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and b) a second nucleotide sequence encoding an antigen from influenza, operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0222] In one example, the self-replicating RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0223] In another example, this disclosure is: (i) Self-replicating RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (b) a second nucleotide sequence encoding an antigen from influenza operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from respiratory syncytial virus (RSV).

[0224] In one example, the self-replicating RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from influenza, operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0225] In one example, the self-replicating RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0226] In another example, this disclosure is: (i) Self-replicating RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide derived from influenza.

[0227] In one example, the self-replicating RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0228] In one example, the self-replicating RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2), operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0229] This disclosure further states that (i) a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0230] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES.

[0231] In one example, the polynucleotide comprises, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0232] In another example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from SARS-CoV-2 operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0233] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0234] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0235] In another example, immunogenic compositions, (i) a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably linked to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0236] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0237] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0238] This disclosure also states, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) We also provide immunogenic compositions containing antigen polypeptides from SARS-CoV-2.

[0239] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES.

[0240] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0241] In another example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0242] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES.

[0243] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0244] In another example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0245] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0246] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0247] This disclosure also states, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES, and (ii) We also provide immunogenic compositions containing antigen polypeptides from SARS-CoV-2.

[0248] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES.

[0249] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0250] In another example, immunogenic compositions, (i) a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0251] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES.

[0252] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0253] In another example, immunogenic compositions, (i) a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0254] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES.

[0255] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0256] This disclosure also states, (i) a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES, and (ii) We also provide immunogenic compositions containing antigen polypeptides from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0257] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES.

[0258] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0259] In another example, immunogenic compositions, (i) a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0260] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES.

[0261] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0262] In another example, immunogenic compositions, (i) a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0263] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES.

[0264] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of a Kozak consensus sequence, IRES, SG promoter, and combinations thereof, and b) a second nucleotide sequence encoding a first antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES.

[0265] This disclosure is, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0266] In one example, the polynucleotide comprises, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0267] In one example, the polynucleotide comprises, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0268] For example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0269] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0270] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0271] For example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0272] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0273] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0274] In another example, this disclosure is: (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0275] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0276] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0277] For example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0278] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0279] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0280] For example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0281] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0282] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0283] In another example, this disclosure is: (i) a cRNA comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0284] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0285] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0286] For example, immunogenic compositions, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0287] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0288] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0289] For example, immunogenic compositions, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0290] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0291] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0292] In another example, this disclosure is: (i) a self-replicating RNA comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0293] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0294] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0295] For example, immunogenic compositions, (i) a self-replicating RNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0296] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0297] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0298] For example, immunogenic compositions, (i) Self-replicating RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0299] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0300] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0301] This disclosure is, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0302] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0303] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0304] For example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0305] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0306] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0307] For example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0308] In one example, the polynucleotide comprises, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0309] In one example, the polynucleotide comprises, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0310] In another example, this disclosure is: (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0311] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0312] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0313] For example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0314] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0315] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0316] For example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0317] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0318] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0319] In another example, this disclosure is: (i) a cRNA comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0320] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0321] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0322] For example, immunogenic compositions, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from SARS-CoV-2 operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0323] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0324] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0325] For example, immunogenic compositions, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from SARS-CoV-2 operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0326] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0327] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0328] In another example, this disclosure is: (i) Self-replicating RNA comprising (a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0329] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0330] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0331] For example, immunogenic compositions, (i) a self-replicating RNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0332] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0333] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0334] For example, immunogenic compositions, (i) Self-replicating RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0335] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0336] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0337] This disclosure is, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0338] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0339] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0340] For example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0341] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0342] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0343] For example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0344] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0345] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0346] In another example, this disclosure is: (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the Kozak consensus sequence and IRES, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0347] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0348] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0349] For example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and IRES, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0350] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0351] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from SARS-CoV-2 operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0352] For example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and IRES, and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0353] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0354] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0355] In another example, this disclosure is: (i) a cRNA comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0356] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0357] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0358] For example, immunogenic compositions, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0359] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0360] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0361] For example, immunogenic compositions, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the Kozak consensus sequence and IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0362] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0363] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0364] In another example, this disclosure is: (i) Self-replicating RNA comprising (a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an IRES, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0365] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0366] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0367] For example, immunogenic compositions, (i) a self-replicating RNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0368] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0369] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the Kozak consensus sequence and the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0370] For example, immunogenic compositions, (i) a self-replicating RNA comprising a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigen polypeptides from influenza.

[0371] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0372] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a Kozak consensus sequence and an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0373] This disclosure is, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0374] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0375] In one example, the polynucleotide comprises, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0376] For example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably linked to an SG promoter, and (b) a second nucleotide sequence encoding an antigen from influenza operably linked to a regulatory element selected from the group consisting of an SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0377] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from SARS-CoV-2 operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0378] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0379] For example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigen polypeptides from influenza.

[0380] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0381] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0382] In another example, this disclosure is: (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an SG promoter, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0383] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0384] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0385] For example, immunogenic compositions, (i) RNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an SG promoter, and b) RNA comprising a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0386] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0387] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0388] For example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the SG promoter, and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES, and (ii) Contains antigen polypeptides from influenza.

[0389] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0390] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0391] In another example, this disclosure is: (i) a cRNA comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0392] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0393] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0394] For example, immunogenic compositions, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0395] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0396] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0397] For example, immunogenic compositions, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigen polypeptides from influenza.

[0398] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0399] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0400] In another example, this disclosure is: (i) a) self-replicating RNA comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0401] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0402] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0403] For example, immunogenic compositions, (i) a) self-replicating RNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0404] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0405] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0406] For example, immunogenic compositions, (i) Self-replicating RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an SG promoter, and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigen polypeptides from influenza.

[0407] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to the SG promoter, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of the SG promoter and IRES.

[0408] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an SG promoter, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0409] This disclosure is, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0410] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0411] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0412] For example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably linked to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0413] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0414] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0415] For example, immunogenic compositions, (i) a polynucleotide comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably linked to an IRES, and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably linked to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigen polypeptides from influenza.

[0416] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0417] In one example, the polynucleotide includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0418] In another example, this disclosure is: (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0419] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0420] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0421] For example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and (b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0422] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0423] In one example, the RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0424] For example, immunogenic compositions, (i) RNA comprising (a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and (b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigen polypeptides from influenza.

[0425] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0426] In one example, the RNA contains, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0427] In another example, this disclosure is: (i) a cRNA comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0428] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0429] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0430] For example, immunogenic compositions, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0431] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0432] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0433] For example, immunogenic compositions, (i) a cRNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigen polypeptides from influenza.

[0434] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0435] In one example, the cRNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0436] In another example, this disclosure is: (i) a) self-replicating RNA comprising a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) To provide an immunogenic composition comprising an antigen polypeptide from severe acute respiratory syndrome coronavirus (SARS-CoV-2).

[0437] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0438] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0439] For example, immunogenic compositions, (i) a self-replicating RNA comprising a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigenic polypeptide from respiratory syncytial virus (RSV).

[0440] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from influenza operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0441] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from influenza operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0442] For example, immunogenic compositions, (i) a self-replicating RNA comprising a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES, and (ii) Contains antigen polypeptides from influenza.

[0443] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0444] In one example, the self-replicating RNA includes, in 5' to 3' order, a) a first nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV) operably ligated to an IRES, and b) a second nucleotide sequence encoding an antigen from severe acute respiratory syndrome coronavirus (SARS-CoV-2) operably ligated to a regulatory element selected from the group consisting of an SG promoter and an IRES.

[0445] For example, this disclosure further states (i) F protein from RSV, and (ii) To provide an immunogenic composition containing sa-mRNA encoding HA from influenza.

[0446] For example, this disclosure further states (i) F protein from RSV, and (ii) To provide an immunogenic composition comprising sa-mRNA encoding HA and NA from influenza.

[0447] For example, this disclosure further states (i) F protein from RSV, and (ii) Provide an immunogenic composition in which each sa-mRNA contains multiple sa-mRNAs encoding HA from influenza.

[0448] For example, this disclosure further states (i) F protein from RSV, and (ii) Provide an immunogenic composition in which each sa-mRNA comprises multiple sa-mRNAs encoding HA and NA from influenza.

[0449] For example, this disclosure further states (i) F protein from RSV, and (ii) Provide an immunogenic composition in which each sa-mRNA contains three or more sa-mRNAs encoding HA from influenza.

[0450] For example, this disclosure further states (i) F protein from RSV, and (ii) Provide an immunogenic composition in which each sa-mRNA comprises three or more sa-mRNAs encoding HA and NA from influenza.

[0451] (i) F protein from RSV, and (ii) Each sa-mRNA is one of four or more sa-mRNAs that encode HA from influenza.

[0452] For example, this disclosure further states (i) F protein from RSV, and (ii) Provide an immunogenic composition in which each sa-mRNA comprises four or more sa-mRNAs encoding HA and NA from influenza.

[0453] In some examples, for instance, when multiple sequences encoding an antigen are contained within a single sa-mRNA, the first nucleotide sequence is operably ligated to the SG promoter, and the second nucleotide sequence is operably ligated to the SG promoter.

[0454] For example, the SG promoter is a natural SG promoter. For instance, the natural SG promoter is a promoter that is natural to and / or based on RNA viruses (e.g., alphaviruses). For example, the natural SG promoter is a natural alphavirus SG promoter.

[0455] In some examples, the SG promoter is coded by a sequence that includes or consists of sequence number 32.

[0456] In one example, the SG promoter is either a minimum SG promoter or an extended SG promoter.

[0457] For example, the SG promoter is the minimal SG promoter. For example, the natural SG promoter is the minimal SG promoter. For example, the minimal SG promoter is the smallest sequence required to initiate transcription. For example, the minimal natural SG promoter is 49 nucleotides long. For example, the minimal SG promoter is 49 nucleotides long. For example, the minimal natural SG promoter is encoded by a sequence containing or consisting of the sequence shown in SEQ ID NO: 13. For example, the minimal SG promoter is encoded by a sequence containing or consisting of the sequence shown in SEQ ID NO: 13.

[0458] In one example, the SG promoter is an extended SG promoter. In another example, the natural SG promoter is an extended SG promoter. For example, the extended SG promoter is extended at its 5' end by nucleotides introduced into the sequence encoding a non-structural protein (e.g., NSP4) of an RNA virus (e.g., alphavirus). In one example, the extended SG promoter is extended at its 5' end by nucleotides introduced into the sequence encoding alphavirus NSP4. The addition of nucleotides to the 5' end of the SG promoter sequence did not interfere with the expression of non-structural proteins and viral replicases, such as alphavirus NSP4.

[0459] In one example, the SG promoter is extended at its 5' end by 51 or fewer nucleotides occurring in a sequence encoding a non-structural protein (e.g., alphavirus NSP4). In another example, the extended SG promoter is a minimal SG promoter extended at its 5' end by 51 or fewer nucleotides occurring in a sequence encoding a non-structural protein (e.g., alphavirus NSP4). In yet another example, the extended SG promoter is encoded by a sequence containing or consisting of the sequence shown in SEQ ID NO: 13, which is extended at its 5' end by 51 or fewer nucleotides occurring in a sequence encoding a non-structural protein (e.g., alphavirus NSP4). For example, the extended SG promoter has a nucleotide length of 100 or fewer. In yet another example, the extended SG promoter is encoded by a sequence containing or consisting of nucleotides 2-101 of SEQ ID NO: 17.

[0460] In one example, the SG promoter is extended at its 5' end by approximately 5 to 20 nucleotides, for example, about 5 nucleotides, or about 10 nucleotides, or about 12 nucleotides, or about 15 nucleotides, or about 20 nucleotides, occurring within the sequence encoding a non-structural protein (e.g., alphavirus NSP4). In another example, the SG promoter is extended at its 5' end by approximately 20 to 35 nucleotides, for example, about 25 nucleotides, or about 27 nucleotides, or about 30 nucleotides, or about 35 nucleotides, occurring within the sequence encoding a non-structural protein (e.g., alphavirus NSP4).

[0461] In one example, the SG promoter is extended at its 5' end by approximately 12 nucleotides that occur within a sequence encoding a non-structural protein (e.g., alphavirus NSP4). In another example, the extended SG promoter is encoded by the sequence shown in SEQ ID NO: 13, which is extended at its 5' end by 12 nucleotides that occur within a sequence encoding a non-structural protein (e.g., alphavirus NSP4). For example, the extended SG promoter has a nucleotide length of 61 or less. In one example, the extended SG promoter is encoded by a sequence containing or consisting of nucleotides 41-101 of SEQ ID NO: 17. In yet another example, the extended SG promoter is encoded by a sequence containing or consisting of the sequence shown in SEQ ID NO: 14.

[0462] In one example, the SG promoter is extended at its 5' end by approximately 31 nucleotides arising from a sequence encoding a non-structural protein (e.g., alphavirus NSP4). In another example, the extended SG promoter is encoded by the sequence shown in SEQ ID NO: 13, which is extended at its 5' end by 31 nucleotides arising from a sequence encoding a non-structural protein (e.g., alphavirus NSP4). For example, the extended SG promoter has a nucleotide length of 80 or less. In one example, the extended SG promoter is encoded by a sequence containing or consisting of nucleotides 22-101 of SEQ ID NO: 17. In yet another example, the extended SG promoter is encoded by a sequence containing or consisting of the sequence shown in SEQ ID NO: 15.

[0463] For example, the extended SG promoter contains a repeating sequence corresponding to nucleotides 66-75 of SEQ ID NO: 17. For instance, the extended SG promoter is encoded by a sequence containing nucleotides 50-75 and nucleotides 66-101 of SEQ ID NO: 17. For example, the extended SG promoter is encoded by the sequence shown in SEQ ID NO: 27.

[0464] For example, IRESs are derived from encephalomyocarditis virus (EMCV), poliovirus (PV), human enterovirus, foot-and-mouth disease virus (FMDV), hepatitis C virus (HCV), classical swine fever virus (CSFV), mouse leukemia virus (MLV), simian immunodeficiency virus (SIV), eukaryotic translation initiation factor 4G (elF4G), death-related protein 5 (DAP5), cellular Myc (c-Myc), NF-κB inhibitor (NRF), vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF-2), platelet-derived growth factor B (PDGF-B), Antennapedia, X-linked inhibitor of apoptosis (XIAP or Apaf-1), immunoglobulin heavy chain binding protein BiP, or fibroblast growth factor 1a (FGF1A), GTX, or combinations thereof.

[0465] For example, IRES is a wild-type IRES derived from encephalomyocarditis virus (EMCV). For instance, wild-type EMCV IRES contains the sequence shown in SEQ ID NO: 16.

[0466] In one example, the nucleotide sequence is codon-optimized.

[0467] In one example, the G / C content of a nucleotide sequence is modified.

[0468] For example, the G / C content of a nucleotide sequence increases by at least 5% compared to the G / C content of an unmodified sequence. For instance, the G / C content of the first and / or second nucleotide sequence, and / or one or more additional nucleotide sequences, increases by at least 10%, 15%, 20%, 25%, 30%, 35%, or 40% compared to the G / C content of an unmodified sequence.

[0469] For example, a polynucleotide contains at least one chemically modified nucleotide.

[0470] For example, the chemically modified nucleotides are selected from the group consisting of N6,2'-O-dimethyladenosine (m6Am), 5-methyluridine (m5U), N4-acetylcytidine (ac4C), 2-thiocytidine (s2C), 2-thiouridine (s2U), 5-methylcytidine (m5C), N6-methyladenosine (m6a), pseudouridine (ψ), 1-methylpseudridine (m1ψ), and combinations thereof. For example, the chemically modified nucleotide is N6,2'-O-dimethyladenosine (m6Am). For example, the chemically modified nucleotide is 5-methyluridine (m5U). For example, the chemically modified nucleotide is N4-acetylcytidine (ac4C). For example, the chemically modified nucleotide is 2-thiocytidine (s2C). For example, a chemically modified nucleotide is 2-thiouridine (s2U). For example, a chemically modified nucleotide is 5-methylcytidine (m5C). For example, a chemically modified nucleotide is N6-methyladenosine (m6a). For example, a chemically modified nucleotide is pseudouridine (ψ). For example, a chemically modified nucleotide is 1-methylpseudridine (m1ψ).

[0471] For example, the nucleotide sequence may include haptoglobin (HP), fibrinogen beta chain (FGB), haptoglobin-related protein (HPR), albumin (ALB), complement component 3 (C3), fibrinogen alpha chain (FGA), alpha-6 collagen (Col6A), alpha-1-antitrypsin (SERPINA1), alpha-1-antichymotrypsin (SERPINA3), its 5'-UTR, fragments, and / or variants.

[0472] In one example, the 5'UTR is the 5'UTR of Venezuelan encephalitis virus (VEEV) or a modified form thereof. For example, the 5'UTR contains the sequence shown in SEQ ID NO: 25.

[0473] For example, 5'-UTR, its fragments, and / or variants have a nucleotide length of 40 to 2000. For example, 5'-UTR, its fragments, and / or variants have a nucleotide length of 40 to 100. For example, 5'-UTR, its fragments, and / or variants have a nucleotide length of 100 to 250. For example, 5'-UTR, its fragments, and / or variants have a nucleotide length of 250 to 500. For example, 5'-UTR, its fragments, and / or variants have a nucleotide length of 500 to 750. For example, 5'-UTR, its fragments, and / or variants have a nucleotide length of 750 to 1000. For example, 5'-UTR, its fragments, and / or variants have a nucleotide length of 1000 to 1250. For example, 5'-UTR, its fragments, and / or variants have a nucleotide length of 1250 to 1500. For example, 5'-UTR, its fragments, and / or variants have a nucleotide length of 1500 to 1750. For example, 5'-UTR, its fragments, and / or variants have a nucleotide length of 1750 to 2000.

[0474] For example, a 5'-UTR, its fragments, and / or variants contain a nucleotide sequence that is at least 90% identical to the nucleotide sequence shown in any one of SEQ ID NOs. For instance, a 5'-UTR, its fragments, and / or variants contain a nucleotide sequence that is 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence shown in any one of SEQ ID NOs.

[0475] In one example, a polynucleotide contains a combination of two or more 5'-UTRs, their fragments, and / or variants. In one example, two or more 5'-UTRs are identical. In another example, two or more 5'-UTRs are different.

[0476] In one example, a nucleotide sequence containing the 5'UTR includes at least one microRNA binding site, an AU-rich element (ARE), a GC-rich element, a stem-loop, and combinations thereof. In one example, the nucleotide sequence includes a microRNA binding site. In one example, the nucleotide sequence includes an AU-rich element (ARE). In one example, the nucleotide includes a GC-rich element. In one example, the nucleotide sequence includes a stem-loop. For example, the stem-loop is a histone stem-loop.

[0477] In one example, a polynucleotide further comprises a nucleotide sequence containing a 3'UTR. In one example, the nucleotide sequence containing the 3'UTR is located at 3' of a second or one or more additional nucleotide sequences. For example, the nucleotide sequence containing the 3'UTR is located at 3' of a second nucleotide sequence. In one example, the 3'UTR includes the arachidonic acid 5-lipoxygenase (ALOX5), alpha-I collagen (COL1A1), tyrosine hydroxylase (TH) gene, the amino-terminal enhancer (AES) of a split, the 3'-UTR of human mitochondrial 12S rRNA (mtRNR1), fragments thereof, and / or variants.

[0478] For example, the 3'UTR is the 3'UTR of Sindbisvirus (SINV) or a modified form thereof. For instance, the 3'UTR contains the sequence shown in SEQ ID NO: 26.

[0479] For example, a 3'UTR, its fragments, and / or variants have a nucleotide length of 40 to 400. For instance, a 3'-UTR has a nucleotide length of 40 to 50, or 50 to 60, or 60 to 70, or 70 to 80, or 80 to 90, or 90 to 100, or 100 to 125, or 125 to 150, or 150 to 175, or 175 to 200, or 200 to 225, or 225 to 250, or 250 to 275, or 275 to 300, or 300 to 325, or 325 to 350, or 350 to 375, or 375 to 400. For example, a 3'-UTR, its fragments, and / or variants have a nucleotide length of 40 to 50. For example, a 3'-UTR, its fragments, and / or variants have a nucleotide length of 50 to 60. For example, 3'-UTR, its fragments and / or variants have a nucleotide length of 60-70. For example, 3'-UTR, its fragments and / or variants have a nucleotide length of 70-80. For example, 3'-UTR, its fragments and / or variants have a nucleotide length of 80-90. For example, 3'-UTR, its fragments and / or variants have a nucleotide length of 90-100. For example, 3'-UTR, its fragments and / or variants have a nucleotide length of 100-125. For example, 3'-UTR, its fragments and / or variants have a nucleotide length of 125-150. For example, 3'-UTR, its fragments and / or variants have a nucleotide length of 150-175. For example, 3'-UTR, its fragments and / or variants have a nucleotide length of 175-200. For example, 3'-UTR, its fragments and / or variants have a nucleotide length of 200-225. For example, 3'-UTR, its fragments, and / or variants have a nucleotide length of 225-250. For example, 3'-UTR, its fragments, and / or variants have a nucleotide length of 250-275. For example, 3'-UTR, its fragments, and / or variants have a nucleotide length of 275-300. For example, 3'-UTR, its fragments, and / or variants have a nucleotide length of 300-325. For example, 3'-UTR, its fragments, and / or variants have a nucleotide length of 325-350.For example, 3'-UTR, its fragments, and / or variants have a nucleotide length of 350 to 375. For example, 3'-UTR, its fragments, and / or variants have a nucleotide length of 375 to 400.

[0480] In one example, a polynucleotide contains a combination of two or more 3'-UTRs, their fragments, and / or variants. In one example, the two or more 3'-UTRs are identical. In another example, the two or more 3'-UTRs are different.

[0481] In one example, a nucleotide sequence containing the 3'UTR, its fragments, and / or variants includes at least one microRNA binding site, an AU-rich element (ARE), a GC-rich element, a triple helix, a stem-loop, one or more stop codons, and combinations thereof. In one example, the nucleotide sequence includes a microRNA binding site. In one example, the nucleotide sequence includes an AU-rich element (ARE). In one example, the nucleotide sequence includes a GC-rich element. In one example, the nucleotide sequence includes a triple helix. In one example, the nucleotide sequence includes a stem-loop. For example, the stem-loop is a histone stem-loop. In one example, the nucleotide sequence includes one or more stop codons. For example, one or more stop codons are located at the 5' end of the 3'-UTR.

[0482] In one example, a polynucleotide comprises a nucleotide sequence containing one or more 3' tailing sequences located at the 3' end of a nucleotide sequence containing a 3' UTR. In one example, one or more 3' tailing sequences are selected from the group consisting of poly-A sequences, polyadenylation signals, G quadruplexes, poly-C sequences, stem-loops, and combinations thereof. For example, the 3' tailing sequence contains a poly-A sequence. In one example, the 3' tailing sequence contains a polyadenylation signal. In one example, the 3' tailing sequence contains a G quadruplex. In one example, the 3' tailing sequence contains a poly-C sequence. In one example, the 3' tailing sequence contains a stem-loop. For example, the stem-loop is a histone stem-loop. In one example, the 3' tailing sequence contains a poly-A sequence and a G quadruplex. In one example, the 3' tailing sequence contains a stem-loop (e.g., a histone stem-loop) and a poly-A sequence.

[0483] For example, one or more 3' tailing sequences contain one or more polyA sequences, each containing 10 to 300 consecutive adenosine nucleotides. For example, each polyA sequence contains 10 to 20, or 20 to 30, or 30 to 40, or 40 to 50, or 50 to 60, or 60 to 70, or 70 to 80, or 80 to 90, or 90 to 100, or 100 to 125, or 125 to 150, or 150 to 175, or 175 to 200, or 200 to 225, or 225 to 250, or 250 to 275, or 275 to 300 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 10 to 20 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 20 to 30 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 30 to 40 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 36 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 40 to 50 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 50 to 60 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 60 to 70 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 70 to 80 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 80 to 90 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 90 to 100 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 100 to 125 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 125 to 150 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 150 to 175 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 175 to 200 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 200 to 225 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 225 to 250 consecutive adenosine nucleotides.For example, one or more polyA sequences each contain 250 to 275 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 275 to 300 consecutive adenosine nucleotides.

[0484] For example, one or more polyA sequences each contain 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, or 300 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 10 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 20 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 30 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 40 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 50 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 60 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 70 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 80 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 90 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 100 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 125 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 150 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 175 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 200 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 225 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 250 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 275 consecutive adenosine nucleotides. For example, one or more polyA sequences each contain 300 consecutive adenosine nucleotides.

[0485] For example, a polyA sequence contains 36 consecutive adenosine nucleotides. For instance, a polyA sequence includes the sequence shown in Sequence ID No. 28.

[0486] In one example, one or more polyA sequences are separated by a break linker. For example, a 3' tailing sequence may contain, from 5' to 3', a polyA sequence containing consecutive adenosine nucleotides, a break linker, and a further polyA sequence containing consecutive adenosine nucleotides.

[0487] For example, a break linker has a nucleotide length of 10-50, 50-100, or 100-150.

[0488] For example, a break linker has a nucleotide length of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, or 150 nucleotides. For example, a break linker has a nucleotide length of 1. For example, a break linker has a nucleotide length of 2. For example, a interrupted linker has a length of 3 nucleotides. For example, a interrupted linker has a length of 4 nucleotides. For example, a interrupted linker has a length of 5 nucleotides. For example, a interrupted linker has a length of 6 nucleotides. For example, a interrupted linker has a length of 7 nucleotides. For example, a interrupted linker has a length of 8 nucleotides. For example, a interrupted linker has a length of 9 nucleotides. For example, a interrupted linker has a length of 10 nucleotides. For example, a interrupted linker has a length of 11 nucleotides. For example, a interrupted linker has a length of 12 nucleotides. For example, a interrupted linker has a length of 13 nucleotides. For example, a interrupted linker has a length of 14 nucleotides. For example, a interrupted linker has a length of 15 nucleotides. For example, a interrupted linker has a length of 16 nucleotides. For example, a interrupted linker has a length of 17 nucleotides. For example, a interrupted linker has a length of 18 nucleotides. For example, a interrupted linker has a length of 19 nucleotides. For example, a interrupted linker has a length of 20 nucleotides. For example, a break linker has a length of 25 nucleotides. For example, a break linker has a length of 30 nucleotides. For example, a break linker has a length of 35 nucleotides. For example, a break linker has a length of 40 nucleotides. For example, a break linker has a length of 45 nucleotides. For example, a break linker has a length of 50 nucleotides.For example, a break linker has a length of 55 nucleotides. For example, a break linker has a length of 60 nucleotides. For example, a break linker has a length of 65 nucleotides. For example, a break linker has a length of 70 nucleotides. For example, a break linker has a length of 75 nucleotides. For example, a break linker has a length of 80 nucleotides. For example, a break linker has a length of 85 nucleotides. For example, a break linker has a length of 90 nucleotides. For example, a break linker has a length of 95 nucleotides. For example, a break linker has a length of 100 nucleotides. For example, a break linker has a length of 110 nucleotides. For example, a break linker has a length of 120 nucleotides. For example, a break linker has a length of 130 nucleotides. For example, a break linker has a length of 140 nucleotides. For example, a break linker has a length of 150 nucleotides.

[0489] In one example, the interrupted linker is 10 nucleotides long. In another example, the interrupted linker contains or consists of the nucleotide sequence shown in Sequence ID No. 20. For example, the interrupted linker contains or consists of the nucleotide sequence GCAUAUGACU.

[0490] In one example, the 3' tailing sequence contains, in order from 5' to 3', a polyA sequence containing 30 consecutive adenosine nucleotides, a 10-nucleotide interruption linker, and a further polyA sequence containing 70 consecutive adenosine nucleotides.

[0491] In one example, the 3' tailing sequence includes, in order from 5' to 3', a polyA sequence containing 30 consecutive adenosine nucleotides, a interruption linker containing or consisting of the nucleotide sequence shown in SEQ ID NO: 20, and a further polyA sequence containing 70 consecutive adenosine nucleotides.

[0492] In one example, RNA further includes a 5' end cap structure.

[0493] For example, the 5' end cap structure is an endogenous cap or an analogue of it.

[0494] For example, the 5'-terminal cap structure contains guanine or a guanine analog.

[0495] For example, the 5'-terminal cap structure is selected from the group consisting of anti-reverse cap analog (ARCA), N7,2'-O-dimethyl-guanosine (mCAP), inosine, N1-methyl-guanosine, 2'-fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, 2-azido-guanosine, N6,2'-O-dimethyladenosine, 7-methylguanosine (m7G), cap 1, and cap 2. For example, the 5'-terminal cap structure is anti-reverse cap analog (ARCA). For example, the 5'-terminal cap structure is N7,2'-O-dimethyl-guanosine (mCAP). For example, the 5'-terminal cap structure is inosine. For example, the 5'-terminal cap structure is N1-methyl-guanosine. For example, the 5'-terminal cap structure is 2'-fluoro-guanosine. For example, the 5' end cap structure is 7-deaza-guanosine. For example, the 5' end cap structure is 8-oxo-guanosine. For example, the 5' end cap structure is 2-amino-guanosine. For example, the 5' end cap structure is LNA-guanosine. For example, the 5' end cap structure is 2-azido-guanosine. For example, the 5' end cap structure is N6,2'-O-dimethyladenosine. For example, the 5' end cap structure is 7-methylguanosine (m7G). For example, the 5' end cap structure is cap 1. For example, the 5' end cap structure is cap 2.

[0496] For example, the 5'-terminal cap structure is ligated to the 5' end of the RNA by a 5'-5'-triphosphate bond or a 5'-5'-phosphorothioate bond.

[0497] In one example, the self-replicating RNA is derived from an alphavirus. For instance, the alphavirus is selected from a group consisting of Semliki Forest Virus (SFV), Sindobis Virus (SIN), Venezuelan Encephalitis Virus (VEE), and combinations thereof.

[0498] In one example, the self-replicating RNA originates from the Semliki Forest Virus (SFV).

[0499] In one example, the self-replicating RNA originates from the Sindbis virus (SIN).

[0500] In one example, the self-replicating RNA originates from the Venezuelan encephalitis virus (VEE).

[0501] For example, self-replicating RNA contains or consists of a polynucleotide sequence as defined in Sequence ID No. 30.

[0502] In one example, the antigen is selected from a group consisting of respiratory syncytial virus (RSV), influenza virus, and / or SARS-CoV-2.

[0503] In one example, the antigen is derived from respiratory syncytial virus (RSV).

[0504] In one example, the antigen is derived from influenza.

[0505] In one example, the antigen is derived from SARS-CoV-2.

[0506] For example, the antigen may be from a single strain (i.e., monovalent) or multiple strains (i.e., polyvalent) of the influenza virus. For instance, an immunogenic composition may contain antigens from one or more (e.g., one, two, or three) influenza virus strains.

[0507] In one example, the composition comprises first and second influenza virus antigens from different strains of influenza virus. In another example, the composition comprises first, second, and third influenza virus antigens from different strains of influenza virus. For example, the composition comprises (i) RNA containing a nucleotide sequence encoding the first influenza antigen, and (ii) polypeptide antigens from different strains of influenza virus. In yet another example, the composition comprises (i) RNA containing a nucleotide sequence encoding the first influenza antigen, (ii) RNA containing a nucleotide sequence encoding the second influenza antigen, and (iii) polypeptide of the third influenza antigen. In yet another example, the composition comprises (i) RNA containing a nucleotide sequence encoding the first influenza antigen from influenza A, (ii) RNA containing a nucleotide sequence encoding the second influenza antigen from influenza B, and (iii) polypeptide of the third influenza antigen from influenza C.

[0508] In one example, the antigen is derived from an influenza A virus strain. For instance, the antigen may be an influenza A virus hemagglutinin (HA) protein, neuraminidase (NA) protein, matrix (M) protein, nucleoprotein (NP), non-structural (NS) protein, or an immunogenic fragment or variant thereof. In one example, the antigen may be an influenza A hemagglutinin (HA) subtype H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15, or H16, and / or an influenza A neuraminidase (NA) subtype N1, N2, N3, N4, N5, N6, N7, N8, or N9, and / or an influenza A matrix (M) protein subtype M1 or M2, and / or an influenza A non-structural (NS) protein subtype NS1 or NS2.

[0509] For example, influenza virus antigens are derived from different subtypes of the influenza virus. For instance, different hemagglutinin subtypes and / or different neuraminidase subtypes and / or matrix protein subtypes and / or nucleoprotein subtypes and / or non-structural protein subtypes.

[0510] Those skilled in the art will recognize that pandemic strains of influenza viruses are generally H1, H2, H3, H5, H6, H7, or H9 subtype influenza A virus strains. For example, H1N1, H2N2, H3N2, H5N1, H5N3, H6N1, H7N2, H7N3, H7N7, H7N9, and H9N2 strains. In one example, the antigen is the H1N1 antigen from the A / Delaware / 55 / 2019 virus strain. In another example, the antigen is the H1 antigen from the A / Delaware / 55 / 2019 virus strain.

[0511] In one example, the antigen is from an influenza A virus strain having the same hemagglutinin subtype. In another example, the antigen is from an influenza A virus strain having a different hemagglutinin subtype. In one example, the antigen is an influenza A virus strain with H1, H2, H3, H5, H6, H7, or H9 subtypes. For example, the antigen is H1 hemagglutinin, or H2 hemagglutinin, or H3 hemagglutinin, or H5 hemagglutinin, or H6 hemagglutinin, or H7 hemagglutinin, or H9 hemagglutinin. For example, the antigen is an influenza A virus strain with H5 subtype (i.e., H5 hemagglutinin). In one example, the H5 hemagglutinin is from the A / turkey / Turkey / 1 / 2005 virus strain. In another example, the H3 hemagglutinin is from the A / Delaware / 39 / 2019 virus strain.

[0512] In one example, the antigen is an influenza A virus strain having the same neuraminidase subtype. In another example, the antigen is an influenza A virus strain having a different neuraminidase subtype. In one example, the antigen is an influenza A virus strain with N1, N2, N3, N7, or N9 subtypes. For example, the antigen is N1 neuraminidase, or N2 neuraminidase, or N3 neuraminidase, or N7 neuraminidase, or N9 neuraminidase. For example, the antigen is an influenza A virus strain with N1 neuraminidase subtype. In one example, the N1 neuraminidase is the A / turkey / turkey / 1 / 2005 strain. In another example, the N2 neuraminidase is the A / Delaware / 39 / 2019 virus strain.

[0513] In one example, the antigens are H5 hemagglutinin protein and / or N1 neuraminidase protein. For instance, the first antigen is an H5 hemagglutinin subtype influenza A virus strain, and the second antigen is an N1 neuraminidase subtype influenza A virus strain.

[0514] In one example, the antigen is an influenza B virus strain. Those skilled in the art will recognize that influenza B viruses are not divided into subtypes, but rather into two lineages, namely B / Yamagata and B / Victoria.

[0515] In one example, the antigen is the B / Yamagata influenza B virus strain. For example, the influenza B virus strain is the B / Singapore / INFTT 16 0610 / 16(By) virus strain. In another example, the antigen is the B / Victoria influenza B virus strain. In one example, the antigen is an influenza B virus strain from the same lineage. In yet another example, the antigen is an influenza B virus strain from a different lineage.

[0516] In one example, the antigen is the influenza B virus Hyam protein and / or Nyam protein. For example, the antigen is the influenza B virus Hyam protein. In another example, the antigen is the influenza B virus Nyam protein. In yet another example, the antigen is the influenza B virus Hyam and Nyam proteins.

[0517] In one example, the antigen is the H1N1 antigen from influenza A, encoded by a sequence that is at least approximately 70%, at least approximately 75%, at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, at least approximately 99%, or 100% identical to SEQ ID NO: 5. In another example, the antigen is the H5N1 antigen from influenza A, encoded by a sequence that is at least approximately 70%, at least approximately 75%, at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, at least approximately 99%, or 100% identical to SEQ ID NO: 7.

[0518] In one example, the antigen is the H1N1 antigen from influenza A and has an amino acid sequence that is at least approximately 70%, at least approximately 75%, at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, at least approximately 99%, or 100% identical to SEQ ID NO: 6. In another example, the antigen is the H5N1 antigen from influenza A and has an amino acid sequence that is at least approximately 70%, at least approximately 75%, at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, at least approximately 99%, or 100% identical to SEQ ID NO: 8.

[0519] In one example, the antigen is derived from influenza B.

[0520] In one example, the antigen is derived from influenza C.

[0521] For example, the antigen may be from a single strain of SARS-CoV-2 (i.e., monovalent) or from multiple strains of SARS-CoV-2 (i.e., polyvalent). For instance, an immunogenic composition may contain antigens from one or more (e.g., one, two, or three) SARS-CoV-2 strains.

[0522] In one example, the composition comprises first and second SARS-CoV-2 antigens from different strains of SARS-CoV-2. In another example, the composition comprises first, second, and third SARS-CoV-2 antigens from different strains of the SARS-CoV-2 virus. For example, the composition comprises (i) RNA containing a nucleotide sequence encoding the first SARS-CoV-2 antigen, and (ii) polypeptide antigens from different strains of SARS-CoV-2. In yet another example, the composition comprises (i) RNA containing a nucleotide sequence encoding the first SARS-CoV-2 antigen, (ii) RNA containing a nucleotide sequence encoding the second SARS-CoV-2 antigen, and (iii) polypeptides of the third SARS-CoV-2 antigen. In another example, the composition comprises (i) RNA containing a nucleotide sequence encoding a first SARS-CoV-2 antigen from the delta strain of SARS-CoV-2, (ii) RNA containing a nucleotide sequence encoding a second SARS-CoV-2 antigen from the omicron strain of SARS-CoV-2, and (iii) a polypeptide of a third SARS-CoV-2 antigen from the Wuhan (original) strain of SARS-CoV-2. In yet another example, the antigen is from one or more of the following SARS-CoV-2 strains: alpha (B.1.1.7), beta (B.1.351), gamma (P1), epsilon (B.1.429), delta (B.1.617.2) mutant, kappa (B.1.617.1) mutant, Wuhan (original) strain, and / or omicron (B.1.1.529) strain.

[0523] In one example, the antigen is the SARS-CoV-2 spike (S) protein and / or nucleocapsid (N) protein. For example, the antigen is the SARS-CoV-2 N protein and / or S protein from the alpha (B.1.1.7), beta (B.1.351), gamma (P1), epsilon (B.1.429), delta (B.1.617.2) mutant, kappa (B.1.617.1) mutant, Wuhan (original) strain and / or omicron (B.1.1.529) strain. In one example, the antigen is the SARS-CoV-2 N protein and / or S protein from the Wuhan (original) SARS-CoV-2 strain. In another example, the antigen is the SARS-CoV-2 N protein and / or S protein from the delta SARS-CoV-2 strain. In one example, the antigen is the SARS-CoV-2 N protein and / or S protein from the delta-SARS-CoV-2 omicron strain.

[0524] In one example, the antigen is the SARS-CoV-2 N protein.

[0525] In another example, the antigen is the extracellular domain of the S protein from the Omicron strain of SARS-CoV-2.

[0526] For example, the antigen is the extracellular domain of the SARS-CoV-2 S protein encoded by a sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to the sequence shown in SEQ ID NO: 9 or 11.

[0527] In another example, the antigen is the extracellular domain of the S protein of the SARS-CoV-2 spike protein, having a sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to the sequence shown in SEQ ID NO: 10 or 12.

[0528] In another example, the S protein is a mutant S protein.

[0529] For example, mutant S proteins contain mutations in the receptor-binding domain. Examples of mutations include: S438F, N439K, N440K, L441I, K444R, V445A, V445I, G446V, G446S, N450K, L452R, L452P, L455F, K458N, N460T, D467V, I468F, I468T, I468V, E471O, I472V, A475V, G476S, S477G, S477I, S477N, S477R, T478I, The group is selected from P479L, P479L, P479S, N481D, N481H, V483F, V483A, E484D, E484K, E484K, E484O, G485S, Y489H, Y489D, Y489F, Y489C, Y489N, F490L, F490S, P491R, Q493L, S494P, Y495N, T500N, N501S, and Y505H, Y508H. In one example, the mutant S protein contains a mutation in the receptor-binding domain selected from the group consisting of N439K, N439L, L452R, S477N, T478I, V483A, and E484D.

[0530] For example, mutant S proteins contain mutations in the receptor-binding domain. Examples of mutations include: R346K, K417N, K417T, S438F, N439K, N440K, L441I, K444R, V445A, V445I, G446V, G446S, N450K, L452R, L452P, L455F, K458N, N460T, D467V, I468F, I468T, I468V, E471O, I472V, A475V, G476S, S477G, S477I, S477N, S477R, The group is selected from T478I, T478K, P479L, P479S, N481D, N481H, V483F, V483A, E484D, E484K, E484K, E484O, G485S, Y489H, Y489D, Y489F, Y489C, Y489N, F490L, F490S, P491R, Q493L, S494P, Y495N, T500N, N501S, N501Y, Y505H, and Y508H. For example, the mutant S protein contains mutations in the receptor-binding domain selected from the group consisting of R346K, K417N, K417T, N439K, N439L, L452R, S477N, T478I, V483A, E484D, E484K, and N501Y.

[0531] For example, mutant S proteins include P337S, F338L, F338C, G339D, E340K, V341I, A344S, T345S, R346K, A348S, A348T, W353R, N354D, N354K, N354S, S359N, D364Y, V367F, S373L, V382L, P384L, P384S, T385A, T393P, V395I, F400C, R403K, R403S, and D4 This includes mutations selected from the group consisting of 05V, R408I, Q414E, Q414K, Q414P, Q414R, T415S, K417R, K417N, I418V, Y421S, Y423C, Y423F, Y423S, D427Y, R509K, V510L, V511E, V512L, L518I, H519O, A520S, A520V, P521R, P521S, A522P, A522S, and D614G.

[0532] For example, mutant S proteins include L18F, D80A, T95I, Y144S, Y145N, D215G, P337S, F338L, F338C, G339D, E340K, V341I, A344S, T345S, R346K, A348S, A348T, W353R, N354D, N354K, N354S, S359N, D364Y, V367F, S373L, V382L, P384L, P384S, T385A, T393P, V39 5I, F400C, R403K, R403S, D405V, R408I, Q414E, Q414K, Q414P, Q414R, T415S, K417N, K417T, K417R, I418V, Y421S, Y423C , Y423F, Y423S, D427Y, S438F, N439K, N440K, L441I, K444R, V445A, V445I, G446V, G446S, N450K, L452R, L452P, L455F, K 458N, N460T, D467V, I468F, I468T, I468V, E471O, I472V, A475V, G476S, S477G, S477I, S477N, S477R, T478I, T478K, P4 79L, P479S, N481D, N481H, V483F, V483A, E484D, E484K, E484K, E484O, G485S, Y489H, Y489D, Y489F, Y489C, Y489N, F490 This includes mutations selected from the group consisting of L, F490S, P491R, Q493L, S494P, Y495N, T500N, N501S, N501Y, Y505H, Y508H, R509K, V510L, V511E, V512L, L518I, H519O, A520S, A520V, P521R, P521S, A522P, A522S, A570D, D614G, P680H, P681H, A701V, T716I, and D950N.

[0533] In one example, the S protein lacks a fulin cleavage site at the S1 / S2 boundary and / or the S2' region.

[0534] For example, a mutant S protein contains mutations (e.g., substitutions) that stabilize the S protein in the pre-fusion conformation. For instance, the S protein contains substitutions K986P and V987P.

[0535] In one example, the antigen may be from a single strain (i.e., monovalent) of respiratory syncytial virus (RSV) or from multiple strains (i.e., polyvalent) of RSV. For example, an immunogenic composition may contain antigens from one or more (e.g., one, two, or three) RSV strains.

[0536] In one example, the composition comprises first and second RSV antigens from different strains of RSV. In another example, the composition comprises first, second, and third RSV antigens from different strains of RSV. For example, the composition comprises (i) RNA containing a nucleotide sequence encoding the first RSV antigen, and (ii) polypeptide antigens from different strains of RSV. In yet another example, the composition comprises (i) RNA containing a nucleotide sequence encoding the first RSV antigen, (ii) RNA containing a nucleotide sequence encoding the second RSV antigen, and (iii) polypeptides of the third RSV antigen from RSV.

[0537] For example, antigens are selected from RSV surface glycoproteins, including fusion proteins (F), glycoproteins (G), small hydrophobic proteins (SH), matrix proteins M and M2, nucleocapsid proteins N, P and L, and non-structural proteins NS1 and NS2. In certain examples, the first and second antigens are RSV-F antigens, respectively. In certain examples, the first, second, and third antigens are RSV-F antigens, respectively.

[0538] In one example, the antigen is from the Pre-F protein of RSV, encoded by a sequence that is at least approximately 70%, at least approximately 75%, at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, at least approximately 99%, or 100% identical to SEQ ID NO: 1 or 3.

[0539] In one example, the antigen is from the Pre-F protein of RSV having a sequence that is at least approximately 70%, at least approximately 75%, at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, at least approximately 99%, or 100% identical to SEQ ID NO: 2 or 4.

[0540] In one example, the antigen is derived from the G protein of RSV.

[0541] In one example, the composition further comprises one or more additional antigens. Thus, in some examples, the composition comprises a third antigen. In another example, the composition comprises a fourth antigen. In one example, one or more additional antigens, or a third or fourth antigen, are viral antigens. For example, the viral antigens are from respiratory viruses. In one example, the respiratory viruses are selected from the group consisting of influenza viruses, respiratory syncytial viruses, parainfluenza viruses, metapneumonia viruses, rhinoviruses, coronaviruses, adenoviruses, and bocaviruses.

[0542] In one example, one or more additional viral antigens are from the influenza virus.

[0543] In one example, one or more additional viral antigens are from respiratory syncytial virus.

[0544] In one example, one or more additional viral antigens are from the parainfluenza virus.

[0545] In one example, one or more additional viral antigens are from metapneumonia virus.

[0546] In one example, one or more additional viral antigens are derived from rhinovirus.

[0547] In one example, one or more additional viral antigens are derived from coronaviruses.

[0548] In one example, one or more additional viral antigens are derived from adenovirus.

[0549] In one example, one or more additional viral antigens are derived from the bocavirus.

[0550] In one example, one or more additional antigens are viral antigens from influenza viruses or coronaviruses.

[0551] In one example, the first, second, third, and / or fourth antigens are expressed at substantially the same level. For example, the antigens have expression levels of about 10%, 5%, or 1% of each other. In another example, the first, second, third, and / or fourth antigens are expressed at different levels. For example, the antigens have expression levels of more than 10%, 15%, or 20% of each other. Methods for determining the levels of expression are known in the art and / or are described herein.

[0552] This disclosure also provides pharmaceutical compositions comprising the immunogenic composition of this disclosure and a pharmaceutically acceptable carrier. Pharmacochemically acceptable carriers suitable for use in this disclosure will be obvious to those skilled in the art and / or are described herein.

[0553] In one example, the pharmaceutical composition further comprises lipid nanoparticles (LNPs), polymer microparticles, and an oil-in-water emulsion. For example, polynucleotides, RNA, cRNA, or self-replicating RNA are encapsulated, bound to, or adsorbed within the LNPs, polymer microparticles, and the oil-in-water emulsion. In one example, polynucleotides are encapsulated, bound to, or adsorbed within the LNPs, polymer microparticles, and the oil-in-water emulsion. In another example, RNA is encapsulated, bound to, or adsorbed within the LNPs, polymer microparticles, and the oil-in-water emulsion. For example, cRNA is encapsulated, bound to, or adsorbed within the LNPs, polymer microparticles, and the oil-in-water emulsion. For example, self-replicating RNA is encapsulated, bound to, or adsorbed within the LNPs, polymer microparticles, and the oil-in-water emulsion.

[0554] In one example, the pharmaceutical composition further comprises LNPs. For example, polynucleotides are encapsulated within the LNPs. In another example, RNA is encapsulated within the LNPs. For example, cRNA is encapsulated within the LNPs. For example, self-replicating RNA is encapsulated within the LNPs. For example, polynucleotides are bound to the LNPs. In another example, RNA is bound to the LNPs. For example, cRNA is bound to the LNPs. In yet another example, self-replicating RNA is bound to the LNPs. For example, polynucleotides are adsorbed onto the LNPs. In yet another example, RNA is adsorbed onto the LNPs. For example, cRNA is adsorbed onto the LNPs. In yet another example, self-replicating RNA is adsorbed onto the LNPs. In yet another example, each RNA is formulated together within the LNPs. In yet another example, each RNA is formulated separately within the LNPs.

[0555] In one example, when multiple RNAs are intended, they are contained within the same LNP. In another example, multiple RNAs are contained within different LNPs.

[0556] For example, this composition, (i) One or more antigens, (ii) One or more immunostimulants, (iii) One or more chemiactive substances, and / or (iv) Further comprising additional RNA encoding one or more target molecules.

[0557] In one example, the additional RNA may be contained within the same LNP as the antigen-coding RNA, or within a separate LNP from the antigen-coding RNA.

[0558] In one example, LNP includes PEG lipids, structural lipids, and / or neutral lipids. For example, LNP includes PEG lipids, structural lipids, and neutral lipids. In another example, LNP includes PEG lipids, structural lipids, or neutral lipids.

[0559] In one example, the LNP further contains cationic lipids. In another example, the LNP does not contain cationic lipids.

[0560] In one example, the pharmaceutical composition further comprises polymer microparticles. For example, polynucleotides are encapsulated within the polymer microparticles. In another example, RNA is encapsulated within the polymer microparticles. For example, cRNA is encapsulated within the polymer microparticles. For example, self-replicating RNA is encapsulated within the polymer microparticles. For example, polynucleotides are bound to polymer microparticles. In another example, RNA is bound to polymer microparticles. For example, cRNA is bound to polymer microparticles. In yet another example, self-replicating RNA is bound to polymer microparticles. For example, polynucleotides are adsorbed onto polymer microparticles. In yet another example, RNA is adsorbed onto polymer microparticles. For example, cRNA is adsorbed onto polymer microparticles. In yet another example, self-replicating RNA is adsorbed onto polymer microparticles.

[0561] In one example, the pharmaceutical composition further comprises an oil-in-water emulsion. For example, polynucleotides are encapsulated in the oil-in-water emulsion. In another example, RNA is encapsulated in the oil-in-water emulsion. For example, cRNA is encapsulated in the oil-in-water emulsion. For example, self-replicating RNA is encapsulated in the oil-in-water emulsion. For example, polynucleotides are bound to the oil-in-water emulsion. In another example, RNA is bound to the oil-in-water emulsion. For example, cRNA is bound to the oil-in-water emulsion. In yet another example, self-replicating RNA is bound to the oil-in-water emulsion. In a further example, self-replicating RNA is adsorbed onto the oil-in-water emulsion. In a further example, self-replicating RNA is resuspended in the oil-in-water emulsion.

[0562] This disclosure also provides immunogenic compositions or pharmaceutical compositions of this disclosure for use as vaccines.

[0563] In one example, the polynucleotide is DNA. In another example, the DNA encodes the cRNA described herein. In yet another example, the DNA encodes the self-replicating RNA vaccine described herein.

[0564] For example, DNA is a plasmid.

[0565] This disclosure provides a method for treating, preventing, or slowing the progression of a disease or condition in a subject, comprising administering the immunogenic composition or pharmaceutical composition of this disclosure to a subject in need. In one example, this disclosure provides a method for treating a disease or condition in a subject, comprising administering the immunogenic composition or pharmaceutical composition of this disclosure to a subject in need. In another example, this disclosure provides a method for preventing a disease or condition in a subject, comprising administering the immunogenic composition or pharmaceutical composition of this disclosure to a subject in need. In yet another example, this disclosure provides a method for slowing the progression of a disease or condition in a subject, comprising administering the immunogenic composition or pharmaceutical composition of this disclosure to a subject in need.

[0566] In one example, the subject suffers from a disease or condition. In another example, the subject has been diagnosed with a disease or condition. In yet another example, the subject is receiving treatment for a disease or condition.

[0567] In one example, the disease or condition is selected from the group consisting of influenza, COVID-19, SARS-CoV-2, respiratory syncytial virus (RSV), and acute respiratory distress syndrome (ARDS), or a combination thereof. In another example, the disease or condition is influenza and COVID-19. In yet another example, the disease or condition is influenza and RSV. In yet another example, the disease or condition is RSV and COVID-19. In yet another example, the disease or condition is influenza, RSV, and COVID-19. In yet another example, the disease or condition is influenza and SARS-CoV-2. In yet another example, the disease or condition is SARS-CoV-2 and RSV. In yet another example, the disease or condition is SARS-CoV-2. In yet another example, the disease or condition is RSV. In yet another example, the disease or condition is influenza. In yet another example, the disease or condition is COVID-19. In yet another example, the disease or condition is ARDS.

[0568] Those skilled in the art will understand that when the treatment, prevention, or delay of the progression of influenza is intended, the immunogenic compositions described herein include at least one polynucleotide comprising a nucleotide sequence encoding an antigen from influenza, or an antigen polypeptide from influenza. The compositions may include additional polynucleotide or polypeptide antigens, for example, from RSV or SARS-CoV-2, as described herein.

[0569] For example, an immunogenic composition suitable for the treatment, prevention, or delaying of the progression of influenza, a) RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0570] In another example, immunogenic compositions suitable for treating, preventing, or delaying the progression of influenza, a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from SARS-CoV-2.

[0571] In another example, immunogenic compositions suitable for treating, preventing, or delaying the progression of influenza, a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0572] In another example, immunogenic compositions suitable for treating, preventing, or delaying the progression of influenza, a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0573] In another example, immunogenic compositions suitable for treating, preventing, or delaying the progression of influenza, a) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0574] In another example, immunogenic compositions suitable for treating, preventing, or delaying the progression of influenza, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from SARS-CoV-2.

[0575] In another example, immunogenic compositions suitable for treating, preventing, or delaying the progression of influenza, (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0576] In another example, immunogenic compositions suitable for treating, preventing, or delaying the progression of influenza, (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0577] In another example, immunogenic compositions suitable for treating, preventing, or delaying the progression of influenza, (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0578] In another example, immunogenic compositions suitable for treating, preventing, or delaying the progression of influenza, (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptide from SARS-CoV-2, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0579] In another example, immunogenic compositions suitable for treating, preventing, or delaying the progression of influenza, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptide from SARS-CoV-2, and (iii) Contains an antigen polypeptide derived from influenza.

[0580] Those skilled in the art will understand that, when the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19 is sought, the immunogenic compositions described herein include at least one polynucleotide containing a nucleotide sequence encoding an antigen from SARS-CoV-2, or an antigen polypeptide from SARS-CoV-2. The compositions may also include additional polynucleotide or polypeptide antigens from, for example, influenza or RSV, as described herein.

[0581] For example, an immunogenic composition suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, a) RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0582] In another example, immunogenic compositions suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from SARS-CoV-2.

[0583] In another example, immunogenic compositions suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, a) RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from SARS-CoV-2.

[0584] In another example, immunogenic compositions suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, a) RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0585] In another example, immunogenic compositions suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, a) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from SARS-CoV-2.

[0586] In another example, immunogenic compositions suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from SARS-CoV-2.

[0587] In another example, immunogenic compositions suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0588] In another example, immunogenic compositions suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0589] In another example, immunogenic compositions suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0590] In another example, immunogenic compositions suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptide from SARS-CoV-2, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0591] In another example, immunogenic compositions suitable for the treatment, prevention, or delay of progression of SARS-CoV-2 infection or COVID-19, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptide from SARS-CoV-2, and (iii) Contains an antigen polypeptide derived from influenza.

[0592] Those skilled in the art will understand that when the treatment, prevention, or delay of progression of RSV is sought, the immunogenic compositions described herein include at least one polynucleotide containing a nucleotide sequence encoding an antigen from RSV, or an antigen polypeptide from RSV. The compositions may include additional polynucleotide or polypeptide antigens, for example, from influenza or SARS-CoV-2, as described herein.

[0593] For example, an immunogenic composition suitable for the treatment, prevention, or delaying of the progression of RSV is: a) RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from RSV

[0594] In another example, immunogenic compositions suitable for the treatment, prevention, or delaying of the progression of RSV are: a) RNA containing a nucleotide sequence encoding an antigen from RSV, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from SARS-CoV-2.

[0595] In another example, immunogenic compositions suitable for the treatment, prevention, or delaying of the progression of RSV are: a) RNA containing a nucleotide sequence encoding an antigen from RSV, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptide from RSV

[0596] In another example, immunogenic compositions suitable for the treatment, prevention, or delaying of the progression of RSV are: a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides from respiratory syncytial virus (RSV).

[0597] In another example, immunogenic compositions suitable for the treatment, prevention, or delaying of the progression of RSV are: a) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Contains antigen polypeptides derived from influenza.

[0598] In another example, immunogenic compositions suitable for the treatment, prevention, or delaying of the progression of RSV are: (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from SARS-CoV-2.

[0599] In another example, immunogenic compositions suitable for the treatment, prevention, or delaying of the progression of RSV are: (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0600] In another example, immunogenic compositions suitable for the treatment, prevention, or delaying of the progression of RSV are: (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains an antigen polypeptide derived from influenza.

[0601] In another example, immunogenic compositions suitable for the treatment, prevention, or delaying of the progression of RSV are: (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0602] In another example, immunogenic compositions suitable for the treatment, prevention, or delaying of the progression of RSV are: (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptide from SARS-CoV-2, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0603] In another example, immunogenic compositions suitable for the treatment, prevention, or delaying of the progression of RSV are: (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptide from SARS-CoV-2, and (iii) Contains an antigen polypeptide derived from influenza.

[0604] This disclosure relates to methods for treating, preventing, or slowing the progression of a disease or condition in a subject, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen selected from the group consisting of respiratory syncytial virus (RSV), influenza, and severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (ii) A method is provided comprising topically co-administering an antigen polypeptide selected from the group consisting of RSV, influenza, and SARS-CoV-2.

[0605] In one example, the subject suffers from a disease or condition. In another example, the subject has been diagnosed with a disease or condition. In yet another example, the subject is receiving treatment for a disease or condition.

[0606] In another example, this method is for preventative purposes, for instance, to prevent or reduce the risk of contracting a disease or condition.

[0607] In one example, the disease or condition is selected from the group consisting of influenza, COVID-19, SARS-CoV-2, respiratory syncytial virus (RSV), and acute respiratory distress syndrome (ARDS), or a combination thereof. In another example, the disease or condition is influenza and COVID-19. In yet another example, the disease or condition is influenza and RSV. In yet another example, the disease or condition is RSV and COVID-19. In yet another example, the disease or condition is influenza, RSV, and COVID-19. In yet another example, the disease or condition is influenza and SARS-CoV-2. In yet another example, the disease or condition is SARS-CoV-2 and RSV. In yet another example, the disease or condition is SARS-CoV-2. In yet another example, the disease or condition is RSV. In yet another example, the disease or condition is influenza. In yet another example, the disease or condition is COVID-19. In yet another example, the disease or condition is ARDS.

[0608] Those skilled in the art will understand that when the treatment, prevention, or delay of the progression of influenza is sought, the methods described herein involve administering at least one polynucleotide or antigen polypeptide from influenza, comprising a nucleotide sequence encoding an antigen from influenza. The composition may include additional polynucleotides or polypeptide antigens from, for example, RSV or SARS-CoV-2, as described herein.

[0609] For example, methods for treating, preventing, or delaying the progression of an infection are: a) RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Including topical co-administration of an antigen polypeptide derived from influenza.

[0610] In another example, this method, a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Including administration of an antigen polypeptide derived from SARS-CoV-2.

[0611] In another example, this method, a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Including administering an antigen polypeptide derived from influenza.

[0612] In another example, this method, a) RNA containing a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) Including administration of an antigen polypeptide derived from respiratory syncytial virus (RSV).

[0613] In another example, this method, a) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and b) Including administering an antigen polypeptide derived from influenza.

[0614] In another example, this method, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Including administering an antigen polypeptide derived from SARS-CoV-2.

[0615] In another example, this method, (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Including administering an antigen polypeptide derived from respiratory syncytial virus (RSV).

[0616] In another example, this method, (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Including administering an antigen polypeptide derived from influenza.

[0617] In another example, this method, (i) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigenic polypeptides from influenza, and (iii) Including administering an antigen polypeptide derived from respiratory syncytial virus (RSV).

[0618] In another example, this method, (i) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptide from SARS-CoV-2, and (iii) Including administering an antigen polypeptide derived from respiratory syncytial virus (RSV).

[0619] In another example, this method, (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) Antigen polypeptide from SARS-CoV-2, and (iii) Including administering an antigen polypeptide derived from influenza.

[0620] Those skilled in the art will understand that the aim is to treat, prevent, or delay the progression of influenza infection and / or RSV infection and / or SARS-CoV-2 infection or COVID-19.

[0621] Preferred combinations of antigens and / or the antigens themselves are described herein with respect to immunogenic compositions and are deemed to be applicable mutatis mutandis to the methods of treatment described herein.

[0622] For example, this disclosure relates to a method for treating, preventing, or delaying the progression of infection caused by influenza and / or RSV. (i) F protein from RSV, and (ii) The present invention provides a method comprising locally co-administering sa-mRNA encoding HA from influenza.

[0623] For example, this disclosure relates to a method for treating, preventing, or delaying the progression of infection caused by influenza and / or RSV. (i) F protein from RSV, and (ii) The present invention provides a method comprising locally co-administering sa-mRNA encoding HA and NA from influenza.

[0624] For example, this disclosure relates to a method for treating, preventing, or delaying the progression of infection caused by influenza and / or RSV. (i) F protein from RSV, and (ii) The present invention provides a method comprising locally and simultaneously administering multiple sa-mRNAs, each sa-mRNA encoding HA from influenza.

[0625] For example, this disclosure relates to a method for treating, preventing, or delaying the progression of infection caused by influenza and / or RSV. (i) F protein from RSV, and (ii) The present invention provides a method comprising locally co-administering multiple sa-mRNAs, each sa-mRNA encoding HA and NA from influenza.

[0626] For example, this disclosure relates to a method for treating, preventing, or delaying the progression of infection caused by influenza and / or RSV. (i) F protein from RSV, and (ii) The present invention provides a method comprising locally co-administering three or more sa-mRNAs, each encoding HA from influenza.

[0627] For example, this disclosure relates to a method for treating, preventing, or delaying the progression of infection caused by influenza and / or RSV. (i) F protein from RSV, and (ii) The present invention provides a method comprising locally co-administering three or more sa-mRNAs, each sa-mRNA encoding HA and NA from influenza.

[0628] (i) F protein from RSV, and (ii) Each sa-mRNA is one of four or more sa-mRNAs that encode HA from influenza.

[0629] For example, this disclosure relates to a method for treating, preventing, or delaying the progression of infection caused by influenza and / or RSV. (i) F protein from RSV, and (ii) The present invention provides a method comprising locally co-administering four or more sa-mRNAs, each sa-mRNA encoding HA and NA from influenza.

[0630] For example, the present disclosure provides a method for inducing an immune response in a subject, which includes administering to a subject in need of such response a pharmaceutical composition disclosed herein, an immunogenic composition disclosed herein, or a vaccine disclosed herein, or carrying out a method described herein.

[0631] For example, this disclosure provides the use of a pharmaceutical composition, an immunogenic composition, or a vaccine disclosed herein in the manufacture of a pharmaceutical product for inducing an immune response in a subject requiring induction of an immune response.

[0632] For example, this disclosure provides a pharmaceutical composition, an immunogenic composition, or a vaccine disclosed herein for use in inducing an immune response in subjects requiring induction of an immune response.

[0633] In one example, the composition or method of the present disclosure induces a humoral immune response in a subject. For example, the humoral immune response is an antibody-mediated immune response. For example, the production of a neutralizing antibody. In another example, the composition induces a cell-mediated immune response. For example, the cell-mediated immune response includes the activation of antigen-specific cytotoxic T cells. For example, the T cells are CD4 T cells and / or CD8 T cells. In one example, the T cells are CD4 T cells. In another example, the T cells are CD8 T cells. In yet another example, the T cells are CD4 and CD8 T cells.

[0634] For example, administration of a pharmaceutical composition disclosed herein, an immunogenic composition disclosed herein, or a vaccine disclosed herein induces a CD4 T cell-mediated immune response.

[0635] For example, administration of a pharmaceutical composition, an immunogenic composition disclosed herein, or a vaccine disclosed herein induces a CD8 T cell-mediated immune response.

[0636] For example, administration of a pharmaceutical composition disclosed herein, an immunogenic composition disclosed herein, or a vaccine disclosed herein induces a CD4 and CD8 T cell-mediated immune response.

[0637] In one example, the CD4 T cell-mediated immune response is a Th0, Th1, and / or Th2 response. For example, the CD4 T cell-mediated immune response is a Th0 response. In another example, the CD4 T cell-mediated immune response is a Th1 response. In yet another example, the CD4 T cell-mediated immune response is a Th2 response. In one example, the CD4 T cell-mediated immune response is a Th0 and Th1 response. In another example, the CD4 T cell-mediated immune response is a Th0 and Th2 response. In yet another example, the CD4 T cell-mediated immune response is a Th1 and Th2 response. In yet another example, the CD4 T cell-mediated immune response is a Th0, Th1, and Th2 response.

[0638] In one example, the CD8 T cell-mediated immune response is a Th0, Th1, and / or Th2 response. For example, the CD8 T cell-mediated immune response is a Th0 response. In another example, the CD8 T cell-mediated immune response is a Th1 response. In yet another example, the CD8 T cell-mediated immune response is a Th2 response. In one example, the CD8 T cell-mediated immune response is a Th0 and Th1 response. In another example, the CD8 T cell-mediated immune response is a Th0 and Th2 response. In yet another example, the CD8 T cell-mediated immune response is a Th1 and Th2 response. In yet another example, the CD8 T cell-mediated immune response is a Th0, Th1, and Th2 response.

[0639] For example, the compositions described herein can modify the Th2:Th1 ratio. For example, the compositions described herein can reduce the Th2:Th1 ratio.

[0640] In one example, the Th0 response cytokine expresses interleukin-2 (IL2+) and / or tumor necrosis factor alpha (TNFa+), and / or is negative for interferon-gamma (IFNg-), IL5-, and / or IL13-. For example, the cytokine is IL2+. In another example, the cytokine is TNFa+. In one example, the cytokine is IFNg-. In yet another example, the cytokine is IL5-. In yet another example, the cytokine is IL13-.

[0641] In one example, the Th1 response cytokine expresses interferon-gamma (IFNg+) and / or is negative for IL5- and / or IL13-. For example, the cytokine is IFNg+. In another example, the cytokine is IL5-. In yet another example, the cytokine is IL13-.

[0642] In one example, the Th2-responsive cytokine expresses IL5+ and / or IL13+ and / or is negative for IFNg. For example, the cytokine is IL5+. In another example, the cytokine is IL13+. For example, the cytokine is IFNg-.

[0643] For example, the present disclosure provides a method for inducing a Th1 immune response in a subject requiring such induction, comprising administering a pharmaceutical composition, an immunogenic composition, or a vaccine disclosed herein to the subject, or carrying out a method described herein.

[0644] For example, this disclosure provides the use of a pharmaceutical composition, an immunogenic composition, or a vaccine disclosed herein in the preparation of a drug for inducing a Th1 immune response in a subject.

[0645] For example, this disclosure provides a pharmaceutical composition, an immunogenic composition, or a vaccine disclosed herein for use in inducing a Th1 immune response in a subject.

[0646] For example, the present disclosure provides a method for reducing the viral load in a subject having a viral infection, the method comprising administering a pharmaceutical composition, an immunogenic composition, or a vaccine disclosed herein to the subject having a viral infection, or carrying out a method described herein.

[0647] For example, this disclosure provides the use of a pharmaceutical composition, an immunogenic composition, or a vaccine disclosed herein in the preparation of a pharmaceutical for reducing the viral load in a subject having a viral infection.

[0648] For example, this disclosure provides a pharmaceutical composition, an immunogenic composition, or a vaccine disclosed herein for use in reducing the viral load in a subject having a viral infection.

[0649] In one example, the subjects are people aged 18 and over. In another example, the subjects are people of any age, for example, from about 1 month to 100 years old, for example, from about 2 months to about 80 years old, from about 6 months to about 3 years old, from about 3 years to about 18 years old, from about 12 years to about 18 years old, from about 18 years to about 55 years old, from about 50 years to about 75 years old, and from about 40 years to about 65 years old. In yet another example, the subjects are people from 2 years old. In yet another example, the subjects are people from 18 years old, 30 years old, 40 years old, 50 years old, 60 years old, 70 years old, 80 years old, or about 90 years old. In yet another example, the subjects are under 2 years old, under 18 months old, under 12 months old, under 6 months old, or under 3 months old.

[0650] In one example, the immunogenic compositions described herein are administered to an infant. In one example, the infant has or is suspected of having RSV, influenza and / or SARS-CoV-2 infection. In one example, the infant is approximately 1 to 12 months old, or 2 to 11 months old, or 3 to 10 months old, or 4 to 9 months old, or 5 to 8 months old, or 6 to 7 months old. In one example, the infant is approximately 1 to 11 months old, approximately 1 to 10 months old, approximately 1 to 9 months old, approximately 1 to 8 months old, approximately 1 to 7 months old, approximately 1 to 6 months old, approximately 1 to 5 months old, approximately 1 to 4 months old, approximately 1 to 3 months old, or approximately 1 to 2 months old. In another example, the infants are approximately 1 month old, 2 months old, 3 months old, 4 months old, 5 months old, 6 months old, 7 months old, 8 months old, 9 months old, 10 months old, 11 months old, or 12 months old.

[0651] Accordingly, one example provided is a method for treating, preventing, or delaying the progression of respiratory syncytial virus (RSV) in an infant, which includes administering an immunogenic composition, pharmaceutical composition, or vaccine disclosed herein to an infant, or carrying out a method described herein.

[0652] Another example provided is the use of immunogenic compositions, pharmaceutical compositions, or vaccines disclosed herein in the preparation of pharmaceuticals for the treatment, prevention, or delay of the progression of respiratory syncytial virus (RSV) in infants.

[0653] In another example, immunogenic compositions, pharmaceutical compositions, or vaccines disclosed herein are provided for use in the treatment, prevention, or delay of the progression of respiratory syncytial virus (RSV) in infants.

[0654] Another example provided is a method for inducing an immune response to respiratory syncytial virus (RSV) in an infant, comprising administering to the infant an immunogenic composition, pharmaceutical composition, or vaccine disclosed herein, or carrying out a method described herein.

[0655] Another example provided is the use of immunogenic compositions, pharmaceutical compositions, or vaccines disclosed herein in the preparation of a drug for inducing an immune response to respiratory syncytial virus (RSV) in infants.

[0656] In another example, an immunogenic composition, pharmaceutical composition, or vaccine disclosed herein is provided for use in inducing an immune response to respiratory syncytial virus (RSV) in infants.

[0657] When the treatment, prevention, or delay of the progression of RSV is sought in infants, those skilled in the art will recognize the preferred immunogenic compositions described herein for administration to infants.

[0658] For example, an immunogenic composition suitable for administration to infants comprises at least one polynucleotide containing a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), and / or an antigen polypeptide from RSV. The composition may also contain additional polynucleotide or polypeptide antigens from, for example, influenza or SARS-CoV-2, as described herein.

[0659] For example, an immunogenic composition suitable for the treatment of RSV in infants is: (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, and (ii) comprising an antigen polypeptide selected from the group consisting of RSV, influenza virus, and / or SARS-CoV-2.

[0660] In another example, an immunogenic composition suitable for the treatment of RSV in infants is: (i) RNA comprising a nucleotide sequence encoding an antigen selected from the group consisting of RSV, influenza virus, and / or SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and (ii) Contains an antigen polypeptide selected from RSV.

[0661] In another example, an immunogenic composition suitable for the treatment of RSV in infants is: (i) RNA comprising a nucleotide sequence encoding an antigen from respiratory syncytial virus (RSV), wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen selected from the group consisting of RSV, influenza, or SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) comprising an antigen polypeptide selected from the group consisting of RSV, influenza, or SARS-CoV-2.

[0662] In another example, an immunogenic composition suitable for the treatment of RSV in infants is: (i) RNA comprising a nucleotide sequence encoding an antigen from influenza or SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, (ii) RNA comprising a nucleotide sequence encoding an antigen from influenza or SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and (iii) Contains antigen polypeptide from respiratory syncytial virus (RSV).

[0663] For example, immunogenic compositions, (i) F protein from RSV, and (ii) Contains sa-mRNA encoding HA from influenza.

[0664] For example, immunogenic compositions, (i) F protein from RSV, and (ii) Contains sa-mRNA encoding HA and NA from influenza.

[0665] For example, immunogenic compositions, (i) F protein from RSV, and (ii) Each sa-mRNA contains multiple sa-mRNAs encoding HA from influenza.

[0666] For example, immunogenic compositions, (i) F protein from RSV, and (ii) Each sa-mRNA contains multiple sa-mRNAs encoding HA and NA from influenza.

[0667] For example, immunogenic compositions, (i) F protein from RSV, and (ii) Each sa-mRNA contains three or more sa-mRNAs encoding HA from influenza.

[0668] For example, immunogenic compositions, (i) F protein from RSV, and (ii) Each sa-mRNA contains three or more sa-mRNAs encoding HA and NA from influenza.

[0669] (i) F protein from RSV, and (ii) Each sa-mRNA is one of four or more sa-mRNAs that encode HA from influenza.

[0670] For example, immunogenic compositions, (i) F protein from RSV, and (ii) Each sa-mRNA contains four or more sa-mRNAs encoding HA and NA from influenza.

[0671] In one example, the composition or vaccine described herein is administered in a single-dose regimen. In another example, the composition is administered in two, three, or four-dose regimens, with doses administered at intervals of approximately one, two, or three months.

[0672] This disclosure also provides polynucleotides encoding self-replicating RNA as described herein. For example, a polynucleotide is recombinant DNA. In one example, recombinant DNA is a plasmid. In one example, the plasmid contains the sequence shown in SEQ ID NO: 31.

[0673] The Disclosure also provides a kit comprising, optionally, at least one immunogenic composition of the Disclosure in a delivery system and / or a pharmaceutically acceptable carrier or diluent, packaged with instructions for use in the treatment, prevention, or delay of the progression of a disease or disorder in a subject.

[0674] The Disclosure also provides a kit comprising, optionally, at least one immunogenic composition of the Disclosure in a delivery system and / or a pharmaceutically acceptable carrier or diluent, packaged with instructions for administering the immunogenic composition to a subject suffering from or at risk of suffering from a disease or disorder.

[0675] In one example, the immunogenic composition, pharmaceutical composition, or vaccine of the Disclosure is supplied in a vial. In another example, the immunogenic composition, pharmaceutical composition, or vaccine of the Disclosure is supplied in a syringe.

[0676] Any discussion of documents, acts, materials, devices, articles, etc., contained herein shall not be deemed to acknowledge that any or all of these matters constitute common knowledge in the art relating to this disclosure, such as forming part of the foundation of the prior art or existing prior to the respective priority dates of the attached claims. [Brief explanation of the drawing]

[0677] [Figure 1] A shows the effect of sa-mRNA on the neutralizing titers of adjuvanted protein vaccines MN and HAI. MN titers were determined by a fluorescence-focused assay (FFA) microneutralization test on MDCK cells. GMT titers represent EC50 values ​​from nonlinear regression with a 95% confidence interval. B shows the effect of sa-mRNA on the titer of adjuvanted protein vaccine IgG. IgG titers were determined by indirect ELISA against monobulk-H1N1:A / Delaware / 55 / 2019. Geometric mean titers for each group are shown. C shows the dose-effect of sa-mRNA on IgG subclasses. IgG1 and IgG2a titers were determined by indirect ELISA against monobulk-H1N1:A / Delaware / 55 / 2019. Geometric mean titers for each group are shown. D shows the effect of sa-mRNA on the IgG1:IgG2 ratio. [Figure 1-1] Same as above. [Figure 2] A shows the effect of sa-mRNA on CD4 T cell cytokines at day 42 in BALB / c mice. B shows the effect of sa-mRNA on CD8 T cell cytokines at day 42 in BALB / c mice. Peptide A is IYSTVASSL influenza A hemagglutinin epitope ID 29690. [Figure 3] A shows the effect of sa-mRNA on the neutralizing titer of adjuvanted protein vaccine MN. B shows the effect of sa-mRNA on the titer of adjuvanted protein vaccine IgG. C shows the dose effect of sa-mRNA on IgG subclasses. D shows the effect of sa-mRNA on the IgG1:IgG2a ratio. [Figure 3-1] Same as above. [Figure 3-2] Same as above. [Figure 4] A shows the effect of sa-mRNA on the Th response. B shows the effect of sa-mRNA on the CD4 T cell response. C shows the effect of sa-mRNA on the CD8 T cell response. D shows the effect of sa-mRNA on the T helper cell response. [Figure 4-1] Same as above. [Figure 4-2] Same as above. [Figure 4-3] Same as above. [Figure 5] A shows the effect of sa-mRNA on influenza MN, NA, and IgG titers. B shows the effect of sa-mRNA on influenza IgG1 and IgG2a responses. C shows the effect of sa-mRNA on omicron MN titer and ACE2 binding. D shows the effect of sa-mRNA on omicron IgG1 and IgG2a responses. [Figure 5-1] Same as above. [Figure 5-2] Same as above. [Figure 5-3] Same as above. [Figure 6]A shows the effect of sa-mRNA and protein-based vaccines on the CD4 T cell response. B shows the effect of sa-mRNA on the T helper cell response. When the ratio was set to 50, 0 Th1 cells were detected. If no response / low response was detected, the S protein ratio was excluded. C shows the effect of sa-mRNA on the CD8 T cell response. [Figure 6-1] Same as above. [Figure 7] A shows the effect of sa-mRNA and protein-based vaccines on the H1N1-specific IgG response. B shows the effect of sa-mRNA on IgG subclasses. C shows the effect of sa-mRNA on H1N1 HAI titer. D shows the effect of sa-mRNA on H1N1 MN titer. E shows the effect of sa-mRNA on N1H1 NAI titer. [Figure 7-1] Same as above. [Figure 7-2] Same as above. [Figure 7-3] Same as above. [Figure 8] A shows the effect of sa-mRNA and protein-based vaccines on neutralization titer in the presence of LNP adjuvants. B shows the effect of sa-mRNA and protein-based vaccines on the Pre-F specific IgG1 response in the presence of LNP adjuvants. C shows the effect of sa-mRNA and protein-based vaccines on the Pre-F specific IgG2a response in the presence of LNP adjuvants. D-E show the effect of sa-mRNA and protein-based vaccines on the Pre-F specific IgG1:IgG2a ratio in the presence of LNP adjuvants. F shows the effect of sa-mRNA and protein-based vaccines on neutralization titer in the presence of LNP adjuvants. [Figure 8-1] Same as above. [Figure 8-2] Same as above. [Figure 9] A represents the effect of sa-mRNA and protein-based vaccines on the Th1 response. [Figure 10]A shows the effect of sa-mRNA on H1N1 HAI, FFA, and IgG titers. B-C show the effect of sa-mRNA on the B subunit antibody response. [Figure 10-1] Same as above. [Figure 10-2] Same as above. [Figure 11] A is the effect of sa-mRNA on the nSP1 Th1 response. B is the effect of sa-mRNA on the H1 Th1 response. C is the effect of sa-mRNA on the B / Yamagata Th2 response. D is the ratio of Th2 / Th1 bias due to the antigen. [Figure 11-1] Same as above. [Figure 11-2] Same as above. [Figure 12A] Includes a series of figures. Shows study designs investigating co-formulation and topical or distal co-administration of sa-mRNA and non-adjuvant or adjuvant-treated protein vaccines. [Figure 12B] The effects of non-adjuvant or adjuvant-treated proteins on the neutralizing antibody response of sa-mRNA vaccines in the context of co-formulation, local co-administration, and distal co-administration were determined by influenza HAI, NAI, and FFA-based MN assays. [Figure 12C] The effects of non-adjuvant or adjuvant-treated proteins on the neutralizing antibody response of sa-mRNA vaccines in the context of co-formulation, local co-administration, and distal co-administration were determined by influenza HAI, NAI, and FFA-based MN assays. [Figure 12D] The effects of non-adjuvant or adjuvant-treated proteins on the neutralizing antibody response of sa-mRNA vaccines in the context of co-formulation, local co-administration, and distal co-administration were determined by influenza HAI, NAI, and FFA-based MN assays. [Figure 12E]The study includes a series of figures. The effect of sa-mRNA on neutralizing antibody responses to non-adjuvant or adjuvant-treated protein vaccines in the context of co-formulation, topical co-administration, and distal co-administration was determined by RSV FFA-based MN assays. [Figure 12F] The effects of co-formulation and topical or distal co-administration on IgG subclass responses were measured by influenza or RSV-conjugated IgG1 and IgG2a ELISA, and the data are presented as the ratio of antigen-specific IgG1 to IgG2a titers. [Figure 12G] The effects of co-formulation and topical or distal co-administration on IgG subclass responses were measured by influenza or RSV-conjugated IgG1 and IgG2a ELISA, and the data are presented as the ratio of antigen-specific IgG1 to IgG2a titers.

[0678] [Table 1] [Modes for carrying out the invention]

[0679] overview Throughout this specification, unless otherwise specifically stated or the context requires, any reference to a single step, composition, group of steps, or group of compositions shall be construed as encompassing one or more (i.e., one or more) of those steps, compositions, groups of steps, or groups of compositions.

[0680] Those skilled in the art will understand that this disclosure is subject to variations and modifications other than those specifically described. It should be understood that this disclosure includes all such variations and modifications. This disclosure also includes, individually or collectively, all of the steps, features, compositions, and compounds referred to or indicated herein, as well as any and all combinations or any two or more of such steps or features.

[0681] This disclosure is not limited in scope by the specific examples described herein, which are for illustrative purposes only. Functionally equivalent products, compositions, and methods are clearly within the scope of this disclosure.

[0682] Any example in this disclosure shall apply mutatis mutandis to any other example in this disclosure unless otherwise specified. In other words, any specific example in this disclosure may be combined with any other specific example in this disclosure (unless they are mutually exclusive).

[0683] Any example in this disclosure disclosing specific features or sets of features or methods or method steps would be construed as providing express support for rejecting those specific features or sets of features or methods or method steps.

[0684] Unless specifically defined otherwise, all technical and scientific terms used herein shall be construed to have the same meaning as those commonly understood by those skilled in the art (e.g., in cell culture, molecular genetics, immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0685] Unless otherwise indicated, the recombinant proteins, cell cultures, and immunological techniques used in this disclosure are standard procedures well known to those skilled in the art. Such techniques are described in J. Perbal, A Practical Guide to Molecular Cloning, John Wiley and Sons (1984), J. Sambrook et al. Molecular Cloning: A Laboratory Manual, Cold Spring Harbour Laboratory Press (1989), TA Brown (editor), Essential Molecular Biology: A Practical Approach, Volumes 1 and 2, IRL Press (1991), DMGlover and BDHames (editors), DNA Cloning: A Practical Approach, Volumes 1-4, IRL Press (1995 and 1996), and FMAusubel et al. (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates to date), Ed Harlow and David Lane (editors), Antibodies: A Laboratory Manual, Cold Spring Harbour Laboratory, (1988), and JEColigan et al. This is described and explained through literature from sources such as al. (editors) Current Protocols in Immunology, John Wiley & Sons (including all updates to date).

[0686] The term "and / or," for example, "X and / or Y," is understood to mean "X and Y" or "X or Y," and is considered to provide explicit support for both meanings or either of them.

[0687] Throughout this specification, the word “comprise,” or variations such as “comprises” or “comprising,” will be understood to mean that they include the elements, elements, or steps, or groups of elements, elements, or steps described herein, but not that they exclude any other elements, elements, or steps, or groups of elements, elements, or steps.

[0688] Where used herein, the term “derived from” shall be interpreted as indicating that a particular integer may be derived from a particular source, but not necessarily directly from that source. Similarly, the term “based on” shall be interpreted as indicating that a particular integer may be developed or used from a particular source, but not necessarily directly from that source.

[0689] Selected definition As used herein, the term “self-replicating RNA” refers to constructs based on RNA viruses that have been engineered to enable the expression of heterologous RNA and proteins. Self-replicating RNA (e.g., in the form of naked RNA) can be amplified in host cells, resulting in the expression of desired gene products in those host cells.

[0690] As used herein, the terms “conventional mRNA,” “cRNA,” or “non-amplified RNA” refer to RNA that is a construct enabling the expression of heterologous RNA and proteins, but which cannot be amplified within a host cell.

[0691] As used herein, the term “monocistronic” with respect to self-replicating RNA refers to RNA that encodes a single polypeptid...

Claims

1. An immunogenic composition, a) RNA comprising a nucleotide sequence encoding an antigen selected from the group consisting of respiratory syncytial virus (RSV), influenza virus, and severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and b) An immunogenic composition comprising an antigen polypeptide selected from the group consisting of RSV, influenza virus, and SARS-CoV-2.

2. a) RNA comprising a nucleotide sequence encoding an antigen from RSV, wherein the nucleotide sequence is operably linked to a regulatory element, and b) The immunogenic composition according to claim 1, comprising an antigen polypeptide from SARS-CoV-2.

3. a) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and b) The immunogenic composition according to claim 1, comprising an antigen polypeptide from RSV.

4. a) RNA comprising a nucleotide sequence encoding an antigen from SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element, and b) The immunogenic composition according to claim 1, comprising an antigen polypeptide from influenza.

5. a) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) The immunogenic composition according to claim 1, comprising an antigen polypeptide from SARS-CoV-2.

6. a) RNA comprising a nucleotide sequence encoding an antigen from influenza, wherein the nucleotide sequence is operably linked to a regulatory element, and b) The immunogenic composition according to claim 1, comprising an antigen polypeptide from RSV.

7. a) RNA comprising a nucleotide sequence encoding an antigen from RSV, wherein the nucleotide sequence is operably linked to a regulatory element, and b) The immunogenic composition according to claim 1, comprising an antigen polypeptide from influenza.

8. The immunogenic composition according to claim 1, further comprising a second RNA comprising a nucleotide sequence encoding an antigen selected from the group consisting of RSV, influenza virus, and SARS-CoV-2, wherein the nucleotide sequence is operably linked to a regulatory element.

9. The immunogenic composition according to claim 1, further comprising a second antigen polypeptide selected from the group consisting of RSV, influenza virus, and SARS-CoV-2.

10. The immunogenic composition according to claim 2 or 3, further comprising a second RNA containing a nucleotide sequence encoding an antigen from influenza operably linked to a regulatory element.

11. From 5' to 3', (i) RNA containing a nucleotide sequence encoding an antigen from RSV or SARS-CoV-2, (ii) The immunogenic composition according to claim 10, comprising RNA containing a nucleotide sequence encoding an antigen from influenza.

12. From 5' to 3', (i) RNA containing a nucleotide sequence encoding an antigen from influenza, and (ii) The immunogenic composition according to claim 10, comprising RNA containing a nucleotide sequence encoding an antigen from RSV or SARS-CoV-2.

13. The immunogenic composition according to claim 2 or 3, further comprising a second antigenic polypeptide from influenza.

14. The immunogenic composition according to claim 4 or 5, further comprising a second RNA containing a nucleotide sequence encoding an antigen from RSV operably linked to a regulatory element.

15. From 5' to 3', (i) RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2, (ii) The immunogenic composition according to 14, comprising RNA containing a nucleotide sequence encoding an antigen from RSV.

16. From 5' to 3', (i) RNA containing a nucleotide sequence encoding an antigen from RSV, and (ii) The immunogenic composition according to 14, comprising RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2.

17. The immunogenic composition according to claim 4 or 5, further comprising a second antigen polypeptide from RSV.

18. The immunogenic composition according to claim 6 or 7, further comprising a second RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2 operably linked to a regulatory element.

19. From 5' to 3', (i) RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2, (ii) The immunogenic composition according to claim 18, comprising RNA containing a nucleotide sequence encoding an antigen from influenza.

20. From 5' to 3', (i) RNA containing a nucleotide sequence encoding an antigen from influenza, and (ii) The immunogenic composition according to claim 18, comprising RNA containing a nucleotide sequence encoding an antigen from SARS-CoV-2.

21. The immunogenic composition according to claim 6 or 7, further comprising a second antigen polypeptide from SARS-CoV-2.

22. The immunogenic composition according to any one of claims 8, 10-12, 14-16, or 18-20, comprising multiple RNAs, each self-replicating RNA encoding a different polypeptide sequence.

23. The immunogenic composition according to any one of claims 1 to 22, wherein the regulatory element is selected from the group consisting of a promoter, an internal ribosome entry site (IRES), a Kozak consensus sequence, or a combination thereof.

24. The immunogenic composition according to claim 23, wherein the promoter is a subgenome (SG) promoter.

25. The immunogenic composition according to claim 24, wherein the SG promoter is a minimal SG promoter.

26. The immunogenic composition according to claim 24, wherein the SG promoter is an extended SG promoter.

27. The immunogenic composition according to claim 25, wherein the minimum SG promoter is encoded by the sequence shown in SEQ ID NO:

13.

28. The immunogenic composition according to claim 26, wherein the extended SG promoter is encoded by the sequence shown in SEQ ID NO:

14.

29. The immunogenic composition according to claim 23, wherein the nucleotide sequence encoding the antigen is operably linked to an IRES located at 5' with respect to the nucleotide sequence encoding the antigen.

30. The immunogenic composition according to claim 23 or 29, wherein the IRES is an IRES derived from encephalomyocarditis virus (EMCV), poliovirus (PV), human enterovirus, foot-and-mouth disease virus (FMDV), hepatitis C virus (HCV), classical swine fever virus (CSFV), mouse leukemia virus (MLV), simian immunodeficiency virus (SIV), eukaryotic translation initiation factor 4G (elF4G), death-related protein 5 (DAP5), cellular Myc (c-Myc), NF-κB inhibitor (NRF), vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF-2), platelet-derived growth factor B (PDGF-B), Antennapedia, X-linked inhibitor of apoptosis (XIAP or Apaf-1), immunoglobulin heavy chain binding protein BiP, or fibroblast growth factor 1a (FGF1A), GTX, or a combination thereof.

31. The immunogenic composition according to claim 30, wherein the EMCV IRES is a wild-type IRES encoded by the sequence shown in Sequence ID No.

16.

32. The immunogenic composition according to any one of claims 1, 2, 7, 8, 11, 12, or 14-16, wherein the nucleotide sequence encoding the antigen from RSV is identical to sequence number 1 or 3 by at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%.

33. The immunogenic composition according to any one of the claims, wherein the nucleotide sequence encoding the antigen from RSV has the sequence shown in SEQ ID NO: 1 or 3.

34. The immunogenic composition according to any one of claims 1, 3, 4, 8, 12, 15, 16, 18, 19, or 20, wherein the nucleotide sequence encoding the antigen from SARS-CoV-2 is identical to sequence number 9 or 11 by at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%.

35. The immunogenic composition according to any one of claims 1, 3, 4, 8, 12, 15, 16, or 18-20, wherein the nucleotide sequence encoding the antigen from SARS-CoV-2 has the sequence shown in SEQ ID NO: 9 or 11.

36. The immunogenic composition according to any one of claims 1, 5, 6, 8, 10-12, 19, or 20, wherein the nucleotide sequence encoding an antigen from influenza has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% the same sequence as SEQ ID NO: 5 or 7.

37. The immunogenic composition according to any one of claims 1, 5, 6, 8, 10-12, 19, or 20, wherein the nucleotide sequence encoding an antigen from influenza has the sequence shown in SEQ ID NO: 5 or 7.

38. The immunogenic composition according to any one of claims 1, 3, 6, 9, or 17, wherein the antigen polypeptide from RSV has an amino acid sequence identical to at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% of SEQ ID NO: 2 or 4.

39. The immunogenic composition according to any one of claims 1, 3, 6, 9, or 17, wherein the antigen polypeptide from RSV has the amino acid sequence shown in SEQ ID NO: 2 or 4.

40. The immunogenic composition according to any one of claims 1, 2, 5, 9, or 21, wherein the antigen polypeptide from SARS-CoV-2 has an amino acid sequence identical to at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% of SEQ ID NO: 10 or 12.

41. The immunogenic composition according to any one of claims 1, 2, 5, 9, or 21, wherein the antigen polypeptide from SARS-CoV-2 has the amino acid sequence shown in SEQ ID NO: 10 or 12.

42. The immunogenic composition according to any one of claims 1, 4, 7, 9, or 13, wherein the antigen polypeptide from influenza has an amino acid sequence identical to that of SEQ ID NO: 6 or 8 by at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%.

43. The immunogenic composition according to any one of claims 1, 4, 7, 9, or 13, wherein the antigen polypeptide from influenza has the amino acid sequence shown in SEQ ID NO: 6 or 8.

44. The immunogenic composition according to any one of claims 1 to 43, wherein the RNA is selected from the group consisting of cRNA or self-replicating RNA.

45. The immunogenic composition according to claim 44, wherein the self-replicating RNA is optionally selected from the group consisting of Semliki Forest Virus (SFV), Sindobis Virus (SIN), Venezuelan Encephalitis Virus (VEE), and combinations thereof, from an alphavirus.

46. An immunogenic composition according to any one of claims 1 to 45, further comprising an adjuvant.

47. The immunogenic composition according to claim 46, wherein the adjuvant is MF59.

48. (i) One or more antigens, (ii) One or more immunostimulants, (iii) One or more chemoattractants, and / or (iv) The immunogenic composition according to any one of claims 1 to 47, further comprising additional RNAs encoding one or more target molecules, or polypeptides selected therefrom.

49. A pharmaceutical composition comprising an immunogenic composition according to any one of claims 1 to 48 and a pharmaceutically acceptable carrier.

50. The pharmaceutical composition according to claim 49, further comprising lipid nanoparticles (LNPs), polymer microparticles, or an oil-in-water emulsion.

51. The pharmaceutical composition according to claim 50, wherein the RNA is encapsulated in, bound to, or adsorbed to LNPs, polymer microparticles, or an oil-in-water emulsion.

52. The pharmaceutical composition according to any one of claims 49 to 51, wherein each RNA is formulated together in the LNP.

53. The pharmaceutical composition according to any one of claims 49 to 51, wherein each RNA is separately formulated in the LNP.

54. An immunogenic composition according to any one of claims 1 to 48, or a pharmaceutical composition according to any one of claims 49 to 53, for use as a vaccine.

55. A vaccine comprising a pharmaceutical composition according to any one of claims 49 to 53, or an immunogenic composition according to any one of claims 1 to 48.

56. A method for carrying out treatment or prevention or delay of the progression of a disease or condition in a subject, comprising administering to the subject an immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55.

57. Use of an immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55 in the manufacture of a pharmaceutical product for carrying out the treatment or prevention or delay of the progression of a disease or condition in a subject that requires such treatment or prevention or delay of progression.

58. An immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55, for use in carrying out treatment or prevention or delay of progression of a disease or condition in a subject requiring such treatment or prevention or delay of progression.

59. A pharmaceutically acceptable composition, immunogenic composition, or vaccine for the use described in claim 56, the use described in claim 57, or the use described in claim 58, wherein the disease or condition is selected from influenza, RSV, SARS-CoV-2 infection, and / or COVID-19.

60. A method for inducing an immune response in a subject, comprising administering to the subject in need of such response an immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55.

61. Use of an immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55 in the manufacture of a pharmaceutical product for inducing an immune response in a subject that requires such induction.

62. An immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55, for use in subjects requiring induction of an immune response.

63. An immunogenic composition, pharmaceutical composition, or vaccine for the method according to claim 60, the use according to claim 61, or the use according to claim 62, wherein the immune response is a cell-mediated immune response, optionally a Th2 response.

64. A method for reducing the viral load in a subject, comprising administering to the subject in need of such reduction an immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55.

65. The use of an immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55 in the preparation of a pharmaceutical for which a reduction in viral load is required in a subject.

66. An immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55, for use in subjects requiring a reduction in viral load.

67. A method for inducing a Th1 immune response in a subject requiring such induction, comprising administering to the subject a pharmaceutical composition according to any one of claims 49 to 53 or a vaccine according to claim 55.

68. The use of a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55, in the preparation of a pharmaceutical for inducing a Th1 immune response in a subject.

69. A pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55, for use in inducing a Th1 immune response in a subject.

70. An immunogenic composition, pharmaceutical composition, or vaccine for the use described in any one of claims 56, 59, 60, 63, 64, or 67, the use described in any one of claims 57, 59, 61, 63, 65, or 68, or the use described in any one of claims 58, 59, 62, 63, 66, or 69, wherein the subject is a human being 18 years of age or older.

71. A method for treating, preventing, or delaying the progression of respiratory syncytial virus (RSV) in an infant, comprising administering to the infant an immunogenic composition, a pharmaceutical composition thereof, or a vaccine according to any one of claims 1, 2, 3, 6-9, 11, 12, or 14-17.

72. Use of an immunogenic composition according to any one of claims 1, 2, 3, 6-9, 11, 12, or 14-17, or a pharmaceutically active composition or vaccine thereof, in the preparation of a pharmaceutical for treating, preventing, or delaying the progression of respiratory syncytial virus (RSV) in infants.

73. An immunogenic composition according to any one of claims 1, 2, 3, 6-9, 11, 12, or 14-17, or a pharmaceutically active composition thereof, or a vaccine, for use in the treatment, prevention, or delay of the progression of respiratory syncytial virus (RSV) in infants.

74. A method for inducing an immune response in an infant, comprising administering to the infant an immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55.

75. Use of an immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55 in the preparation of a pharmaceutical product for inducing an immune response in an infant.

76. An immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55, for use in inducing an immune response in an infant.

77. A pharmaceutical composition, immunogenic composition, or vaccine for the method according to claim 74, the use according to claim 75, or the use according to claim 76, wherein the immune response is a cell-mediated immune response, optionally a Th1 response.

78. An immunogenic composition, pharmaceutical composition, or vaccine for the method according to claim 71, the use according to claim 72, or the use according to claim 73, wherein the infant is approximately 1 to 12 months old.

79. A pharmaceutically acceptable composition, immunogenic composition, or vaccine for use according to any one of claims 56, 59, 60, 63, 64, 67, 70, 71, 74, or 77, wherein the vaccine or composition is administered in a single-dose regimen; a use according to any one of claims 57, 59, 61, 63, 65, 68, 70, 72, 75, 77, or 78; or a pharmaceutically acceptable composition, immunogenic composition, or vaccine for use according to any one of claims 58, 59, 62, 63, 66, 69, 70, 73, 76-78.

80. The method according to any one of claims 56, 59, 60, 63, 64, 67, 70, 71, 74, or 77, wherein the composition or vaccine is administered in two, three, or four dose regimens, with the doses administered at intervals of about one, two, or three months; the use according to any one of claims 57, 59, 61, 63, 65, 68, 70, 72, 75, 77, or 78; or a pharmaceutical composition, immunogenic composition, or vaccine for the use according to any one of claims 58, 59, 62, 63, 66, 69, 70, 73, 76-78.

81. It's a kit, (a) an immunogenic composition according to any one of claims 1 to 48, a pharmaceutical composition according to any one of claims 49 to 53, or a vaccine according to claim 55, (b) the user manual, and optionally, (c) A kit containing a diluent.

82. A method for treating, preventing, or slowing the progression of a disease or condition in a subject, (i) A polynucleotide comprising a nucleotide sequence encoding an antigen selected from the group consisting of respiratory syncytial virus (RSV), influenza, and severe acute respiratory syndrome coronavirus (SARS-CoV-2), wherein the nucleotide sequence is operably linked to a regulatory element, and (ii) A method comprising topically co-administering an antigen polypeptide selected from the group consisting of RSV, influenza, and SARS-CoV-2.

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