Low-dose vaccine composition

Abstract

The present disclosure provides a vaccine composition comprising six or more homologous distinct antigenic components. Any two of the six or more homologous distinct antigenic components may share 98% or less than 95% sequence identity. The homologous distinct antigenic components may comprise a protein. The amount of protein in a dose of the human adult vaccine composition may be about 600 nanograms (ng) to about 3 micrograms (μg). The homologous distinct antigenic components may comprise a plurality of RNAs. The amount of RNA in the plurality of RNAs in the dose of the vaccine composition may be about 1 ng to about 5 μg per dose. The homologous distinct antigenic components may comprise a plurality of proteins displayed on heterologous virus-like particles (VLPs). The amount of protein displayed on the heterologous VLP among the plurality of proteins displayed on the heterologous VLP in a dose of the vaccine composition may be about 1 ng to about 5 μg per dose.

Description

[Technical Field]

[0001] cross reference This application claims the benefit of U.S. Provisional Application No. 63 / 383,034, filed November 9, 2022, which is incorporated herein by reference. [Background technology]

[0002] Abstract Vaccines may contain multiple homologous antigenic components, particularly for pathogens belonging to diverse phylogenetic families (e.g., influenza). For some antigenic components, there may be a minimal dose of that antigenic component required to elicit an optimal immune response to that antigenic component alone. Summary of the Invention [Means for solving the problem]

[0003] overview The present disclosure provides vaccine compositions comprising a plurality of RNAs collectively encoding six or more homologous distinct antigenic components, wherein any two of the six or more homologous distinct antigenic components share less than 98% sequence identity, and each RNA encoding a distinct antigenic component is present in the composition in an amount of 1 ng to 5 micrograms per dose; a plurality of virus-like particles collectively displaying at least six or more homologous distinct antigenic components, wherein any two of the six or more homologous distinct antigenic components share less than 98% sequence identity, and each distinct antigenic component is present in the composition in an amount of 1 ng to 5 micrograms; or at least six or more homologous distinct antigenic components, wherein any two of the six or more homologous distinct antigenic components share less than 98% sequence identity, and each distinct antigenic component of the six or more homologous distinct antigenic components is present in the composition in an amount of 550 ng to 5 micrograms. In some embodiments, the vaccine composition is for the prevention of influenza. In some embodiments, the vaccine composition is for the prevention of HIV. In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount of 1 ng to 2.5 micrograms per dose. In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount of 1 ng to 1.5 micrograms per dose. In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount of 1 ng to 1 microgram per dose. In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount of about 0.5 micrograms per dose. In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount of 1 microgram per dose. In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount of 2 micrograms per dose. In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount of 1 ng to 4 micrograms per dose.In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount greater than 10 nanograms per dose. In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount greater than 100 nanograms per dose. In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount greater than 250 nanograms per dose. In some embodiments, each RNA encoding a different antigenic component is present in the composition in an amount greater than 500 nanograms per dose. In some embodiments, each different antigenic component displayed on the VLP is present in the composition in an amount of 1 ng to 2.5 micrograms per dose. In some embodiments, each different antigenic component displayed on the VLP is present in the composition in an amount of 1 ng to 1.5 micrograms per dose. In some embodiments, each different antigenic component displayed on the VLP is present in the composition in an amount of 1 ng to 1 microgram per dose. In some embodiments, each different antigenic component displayed on the VLP is present in the composition in an amount of about 0.5 micrograms per dose. In some embodiments, each distinct antigenic component of the six or more homologous different antigenic components is present in the composition in an amount of 550 ng to 2.5 micrograms per dose. In some embodiments, each distinct antigenic component of the six or more homologous different antigenic components is present in the composition in an amount of 550 ng to 1.5 micrograms per dose. In some embodiments, each distinct antigenic component of the six or more homologous different antigenic components is present in the composition in an amount of 550 ng to 1 microgram per dose. In some embodiments, each of the at least six or more homologous different antigenic components is present in the composition in an amount of 550 ng to 3000 ng. In some embodiments, each of the at least six or more homologous different antigenic components is present in the composition in an amount of 600 ng to 3000 ng. In some embodiments, each of the at least six or more homologous different antigenic components is present in the composition in an amount of 750 ng to 3000 ng.In some embodiments, each of the at least six or more homologous distinct antigenic components is present in the composition in an amount of 1000 ng to 3000 ng. In some embodiments, each of the at least six or more homologous distinct antigenic components is present in the composition in an amount of 750 ng to 2000 ng. In some embodiments, the vaccine composition is for administration to a human subject. In some embodiments, the vaccine composition is for administration to adults 18 years of age or older. In some embodiments, the vaccine composition is for administration to adults 25 years of age or older. In some embodiments, the vaccine composition is for administration to adults 50 years of age or older. In some embodiments, the vaccine composition is for administration to adults 75 years of age or older. In some embodiments, the vaccine composition is for administration to children between about 1 day of age and about 18 years of age. In some embodiments, the vaccine composition is for administration to children between about 1 day of age and about 5 years of age. In some embodiments, the vaccine composition is for administration to children between about 5 years of age and about 18 years of age. In some embodiments, the vaccine composition is for administration to an animal. In some embodiments, the animal is a livestock animal. In some embodiments, the livestock animal is a cow, a bull, an alpaca, a llama, a sheep, a pig, or a bird. In some embodiments, the animal is a domestic animal. In some embodiments, the domestic animal is a primate. In some embodiments, the six or more homologous distinct antigenic components comprise antigens from a virus, a bacterium, a fungus, a prion, or a plant. In some embodiments, the homologous distinct antigenic components comprise a viral component. In some embodiments, the viral component is a protein comprising a receptor binding domain. In some embodiments, the viral component is a receptor binding domain. In some embodiments, at least two homologous distinct antigenic components of the six or more homologous distinct antigenic components comprise a receptor binding domain for a cell surface protein. In some embodiments, the cell surface protein is mammalian. In some embodiments, the receptor binding domains of any two homologous distinct antigenic components of the six or more homologous distinct antigenic components share less than 99% sequence identity.In some embodiments, the receptor binding domains of any two of the six or more homologous different antigenic components share less than 98% sequence identity. In some embodiments, the receptor binding domains of any two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of any two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of any two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of any two of the six or more homologous different antigenic components share less than 90% sequence identity. In some embodiments, the receptor binding domains of any two of the six or more homologous different antigenic components share less than 85% sequence identity. In some embodiments, the receptor binding domains of any two of the six or more homologous different antigenic components share less than 80% sequence identity. In some embodiments, the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 75% sequence identity. In some embodiments, the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 70% sequence identity. In some embodiments, administration of the vaccine composition reduces the predominance of immunogenic single variant epitopes corresponding to the homologous distinct antigenic components. In some embodiments, any two of the six or more homologous distinct antigenic components share less than about 98% sequence identity. In some embodiments, any two of the six or more homologous distinct antigenic components share less than about 97% sequence identity. In some embodiments, any two of the six or more homologous distinct antigenic components share less than about 96% sequence identity. In some embodiments, any two of the six or more homologous distinct antigenic components share less than about 95% sequence identity.In some embodiments, any two of the six or more homologous different antigenic components share less than about 96% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 95% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 90% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 85% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 80% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 75% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 70% sequence identity. In some embodiments, the six or more homologous different antigenic components comprise seven or more homologous different antigenic components. In some embodiments, the six or more homologous different antigenic components comprise 10 or more homologous different antigenic components. In some embodiments, the six or more homologous different antigenic components comprise 15 or more homologous different antigenic components. In some embodiments, the six or more homologous different antigenic components comprise 20 or more homologous different antigenic components. In some embodiments, the six or more homologous different antigenic components comprise 25 or more homologous different antigenic components. In some embodiments, the six or more homologous different antigenic components comprise 30 or more homologous different antigenic components. In some embodiments, any two of the six or more homologous different antigenic components share at least 30% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least 40% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least 50% sequence identity.In some embodiments, any two of the six or more homologous distinct antigenic components share at least 60% sequence identity. In some embodiments, any two of the six or more homologous distinct antigenic components share at least 70% sequence identity. In some embodiments, the vaccine composition further comprises an adjuvant. In some embodiments, the plurality of RNAs are present in the vaccine composition in the same amount. In some embodiments, the plurality of RNAs are present in the vaccine composition in different amounts. In some embodiments, the dose of one RNA of the plurality of RNAs is different from the homologous distinct antigenic components of the six or more homologous distinct antigenic components. In some embodiments, the dose of one RNA of the plurality of RNAs is calculated to be proportional to the average phylogenetic distance of a homologous distinct antigenic component of the six or more homologous distinct antigenic components to another homologous distinct antigenic component of the six or more homologous distinct antigenic components.

[0004] Another aspect of the present disclosure provides a method of inducing an immune response to an influenza pathogen in a subject, the method comprising administering any of the vaccine compositions disclosed herein. In some embodiments, the method is for prophylaxis against influenza.

[0005] One aspect of the present disclosure provides a vaccine composition comprising six or more homologous distinct antigenic components, any two of which share less than 95% sequence identity; the homologous distinct antigenic components comprise a protein, and the protein concentration / amount in a dose of the vaccine composition for a human adult is about 1 nanogram (ng) to about 3 micrograms (μg); the homologous distinct antigenic components comprise a plurality of RNAs, and the RNA concentration / amount of the plurality of RNAs in a dose of the vaccine composition is about 1 ng to about 5 μg per dose; or the homologous distinct antigenic components comprise a plurality of proteins displayed on heterologous virus-like particles (VLPs), and the protein concentration / amount of the plurality of proteins displayed on the heterologous VLPs in a dose of the vaccine composition is about 1 ng to about 5 μg per dose. In some embodiments, the vaccine composition is for administration to a human subject. In some embodiments, the vaccine composition is for administration to adults 18 years of age or older. In some embodiments, the vaccine composition is for administration to adults 25 years of age or older. In some embodiments, the vaccine composition is for administration to adults 50 years of age or older. In some embodiments, the vaccine composition is for administration to adults 75 years of age or older. In some embodiments, the vaccine composition is for administration to children between about 1 day of age and about 18 years of age. In some embodiments, the vaccine composition is for administration to children between about 1 day of age and about 5 years of age. In some embodiments, the vaccine composition is for administration to children between about 5 years of age and about 18 years of age. In some embodiments, the vaccine composition is for administration to an animal. In some embodiments, the animal is a livestock animal. In some embodiments, the livestock animal is a cow, a bull, an alpaca, a llama, a sheep, a pig, or a bird. In some embodiments, the animal is a farm animal. In some embodiments, the farm animal is a primate. In some embodiments, the six or more homologous different antigenic components comprise antigens selected from the group consisting of a virus, a bacterium, a fungus, a prion, a plant, or a combination thereof.In some embodiments, the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is from about 1 nanogram (ng) to about 5 micrograms (μg). In some embodiments, the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is from about 1 nanogram (ng) to about 1 microgram (μg). In some embodiments, the homologous distinct antigenic component comprises a viral component. In some embodiments, the viral component is a receptor binding domain. In some embodiments, at least two of the six or more homologous distinct antigenic components comprise a receptor binding domain for a cell surface protein. In some embodiments, the cell surface protein is mammalian. In some embodiments, the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 99% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 98% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 90% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 85% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 80% sequence identity.In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 75% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 70% sequence identity. In some embodiments, administration of the vaccine composition reduces the predominance of immunogenic single variant epitopes corresponding to the homologous different antigenic components. In some embodiments, any two of the six or more homologous different antigenic components share less than about 90% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 85% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 80% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 75% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 70% sequence identity. In some embodiments, the vaccine composition comprises seven or more homologous different antigenic components. In some embodiments, the vaccine composition comprises ten or more homologous different antigenic components. In some embodiments, the vaccine composition comprises fifteen or more homologous different antigenic components. In some embodiments, the vaccine composition comprises twenty or more homologous different antigenic components. In some embodiments, the vaccine composition comprises twenty-five or more homologous different antigenic components. In some embodiments, the vaccine composition comprises thirty or more homologous different antigenic components. In some embodiments, the vaccine composition further comprises one or more heterologous antigenic components. In some embodiments, the vaccine composition further comprises an adjuvant. In some embodiments, the six or more homologous different antigenic components are present in the vaccine composition at the same concentration.In some embodiments, the six or more homologous distinct antigenic components are present in different concentrations / amounts in the vaccine composition. In some embodiments, a single dose of a homologous distinct antigenic component among the six or more homologous distinct antigenic components is calculated to be proportional to the phylogenetic distance of the homologous distinct antigenic component to another homologous distinct antigenic component, where another homologous distinct antigenic component has the smallest distance to the homologous distinct antigenic component among the six or more homologous distinct antigenic components. In some embodiments, a single dose of a homologous distinct antigenic component among the six or more homologous distinct antigenic components is calculated to be proportional to its average phylogenetic distance from other homologous distinct antigenic components among the six or more homologous distinct antigenic components. In some embodiments, the vaccine composition comprises a fragment of a SARS virus. In some embodiments, the vaccine composition comprises a fragment of an influenza virus. In some embodiments, the vaccine composition comprises a fragment of an HIV virus. In some embodiments, the vaccine composition comprises a fragment of a SARS1 virus.

[0006] Another aspect of the present disclosure provides a vaccine composition for human adults comprising six or more homologous distinct antigenic components, two of which share less than 95% sequence identity; the homologous distinct antigenic components comprise proteins, wherein the concentration / amount of protein in a dose of the human adult vaccine composition is about 1 nanogram (ng) to about 3 micrograms (μg); the homologous distinct antigenic components comprise a plurality of RNAs, wherein the concentration / amount of RNA of the plurality of RNAs in a dose of the human adult vaccine composition is about 1 ng to about 5 μg per dose; or the homologous distinct antigenic components comprise a plurality of proteins displayed on heterologous virus-like particles (VLPs), wherein the concentration / amount of protein displayed on the heterologous VLP of the plurality of proteins displayed on the heterologous VLP in a dose of the human adult vaccine composition is about 1 ng to about 5 μg per dose. In some embodiments, the vaccine composition is for administration to adults 25 years of age or older. In some embodiments, the vaccine composition is for administration to adults 50 years of age or older. In some embodiments, the vaccine composition is for administration to adults 75 years of age or older. In some embodiments, the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is about 1 nanogram (ng) to about 2 μg. In some embodiments, the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is about 1 nanogram (ng) to about 1 microgram (μg). In some embodiments, the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is about 1 nanogram (ng) to about 0.1 microgram (μg). In some embodiments, the homologous distinct antigenic component comprises a viral component. In some embodiments, the viral component is a receptor binding domain. In some embodiments, at least two of the six or more homologous distinct antigenic components comprise receptor binding domains for cell surface proteins. In some embodiments, the cell surface protein is mammalian.In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 99% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 98% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 90% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 85% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 80% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 75% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 70% sequence identity. In some embodiments, administration of the vaccine composition reduces the predominance of immunogenic single variant epitopes corresponding to the homologous different antigenic components. In some embodiments, any two of the six or more homologous different antigenic components share less than about 90% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 85% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 80% sequence identity.In some embodiments, any two of the six or more homologous different antigenic components share less than about 75% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 70% sequence identity. In some embodiments, the vaccine composition comprises seven or more homologous different antigenic components. In some embodiments, the vaccine composition comprises ten or more homologous different antigenic components. In some embodiments, the vaccine composition comprises fifteen or more homologous different antigenic components. In some embodiments, the vaccine composition comprises twenty or more homologous different antigenic components. In some embodiments, the vaccine composition comprises twenty-five or more homologous different antigenic components. In some embodiments, the vaccine composition comprises thirty or more homologous different antigenic components. In some embodiments, any two of the six or more homologous different antigenic components share at least about 25% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 30% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 35% sequence identity. In some embodiments, the vaccine composition further comprises one or more non-homologous antigenic components. In some embodiments, the vaccine composition further comprises an adjuvant. In some embodiments, the six or more homologous different antigenic components are present in the vaccine composition at the same concentration / amount. In some embodiments, the six or more homologous different antigenic components are present in the vaccine composition at different concentrations / amounts. In some embodiments, a dose of a homologous different antigenic component among the six or more homologous different antigenic components is calculated to be proportional to the phylogenetic distance between the homologous different antigenic component and another homologous different antigenic component, wherein the other homologous different antigenic component has the smallest distance to the homologous different antigenic component among the six or more homologous different antigenic components.In some embodiments, a dose of a homologous distinct antigenic component of the six or more homologous distinct antigenic components is calculated to be proportional to its average phylogenetic distance from other homologous distinct antigenic components of the six or more homologous distinct antigenic components. In some embodiments, the vaccine composition comprises a fragment of a SARS virus. In some embodiments, the vaccine composition comprises a fragment of an influenza virus. In some embodiments, the vaccine composition comprises a fragment of an HIV virus. In some embodiments, the vaccine composition comprises a fragment of a SARS1 virus.

[0007] Another aspect of the present disclosure provides a vaccine composition comprising six or more homologous distinct antigenic components, two of which share less than 95% sequence identity; the homologous distinct antigenic components comprise a protein, wherein the protein concentration / amount in a dose of the vaccine composition is about 1 nanogram (ng) to about 1 microgram (μg); the homologous distinct antigenic components comprise a plurality of RNAs, wherein the RNA concentration / amount in a dose of the vaccine composition is about 1 ng to about 2.5 μg per dose; or the homologous distinct antigenic components comprise a plurality of proteins displayed on heterologous virus-like particles (VLPs), wherein the protein concentration / amount in a dose of the vaccine composition is about 1 ng to about 2.5 μg per dose; and the vaccine composition is a human pediatric vaccine composition. In some embodiments, the vaccine composition is for administration to children between about 1 day old and about 18 years old. In some embodiments, the vaccine composition is for administration to children between about 1 day old and about 5 years old. In some embodiments, the vaccine composition is for administration to children between about 5 and about 18 years of age. In some embodiments, the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is between about 1 nanogram (ng) and about 0.5 micrograms (μg). In some embodiments, the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is between about 1 nanogram (ng) and about 0.1 micrograms (μg). In some embodiments, the homologous distinct antigenic components comprise a viral component. In some embodiments, the viral component is a receptor binding domain. In some embodiments, at least two of the six or more homologous distinct antigenic components comprise a receptor binding domain for a cell surface protein. In some embodiments, the cell surface protein is mammalian. In some embodiments, the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 99% sequence identity.In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 98% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 90% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 85% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 80% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 75% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 70% sequence identity. In some embodiments, administration of the vaccine composition reduces the predominance of immunogenic single variant epitopes corresponding to the homologous different antigenic components. In some embodiments, any two of the six or more homologous different antigenic components share less than about 90% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 85% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 80% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 75% sequence identity.In some embodiments, any two of the six or more homologous different antigenic components share less than about 70% sequence identity. In some embodiments, the vaccine composition comprises seven or more homologous different antigenic components. In some embodiments, the vaccine composition comprises ten or more homologous different antigenic components. In some embodiments, the vaccine composition comprises fifteen or more homologous different antigenic components. In some embodiments, the vaccine composition comprises twenty or more homologous different antigenic components. In some embodiments, the vaccine composition comprises twenty-five or more homologous different antigenic components. In some embodiments, the vaccine composition comprises thirty or more homologous different antigenic components. In some embodiments, any two of the six or more homologous different antigenic components share at least about 25% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 30% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 35% sequence identity. In some embodiments, the vaccine composition further comprises one or more heterologous antigenic components. In some embodiments, the vaccine composition further comprises an adjuvant. In some embodiments, the six or more homologous different antigenic components are present in the vaccine composition at the same concentration. In some embodiments, the six or more homologous different antigenic components are present in the vaccine composition at different concentrations / amounts. In some embodiments, a dose of a homologous different antigenic component among the six or more homologous different antigenic components is calculated to be proportional to the phylogenetic distance between the homologous different antigenic component and another homologous different antigenic component, where the other homologous different antigenic component has the smallest distance to the homologous different antigenic component among the six or more homologous different antigenic components. In some embodiments, a dose of a homologous different antigenic component among the six or more homologous different antigenic components is calculated to be proportional to its average phylogenetic distance from other homologous different antigenic components among the six or more homologous different antigenic components.In some embodiments, the vaccine composition comprises a fragment of a SARS virus. In some embodiments, the vaccine composition comprises a fragment of an influenza virus. In some embodiments, the vaccine composition comprises a fragment of an HIV virus. In some embodiments, the vaccine composition comprises a fragment of a SARS1 virus.

[0008] Another aspect of the present disclosure provides a vaccine composition comprising six or more homologous distinct antigenic components, wherein two of the six or more homologous distinct antigenic components share less than 95% sequence identity, the homologous distinct antigenic components comprise proteins, wherein the concentration / amount of protein in a dose of the vaccine composition is about 1 nanogram (ng) to about 3 micrograms (μg), the homologous distinct antigenic components comprise a plurality of RNAs, wherein the concentration / amount of RNA of the plurality of RNAs in a dose of the vaccine composition is about 1 ng to about 5 μg per dose, or the homologous distinct antigenic components comprise a plurality of proteins displayed on heterologous virus-like particles (VLPs), wherein the concentration / amount of protein displayed on the heterologous VLP of the plurality of proteins displayed on the heterologous VLP in a dose of the vaccine composition is about 1 ng to about 5 μg per dose, and the vaccine composition is a veterinary vaccine composition. In some embodiments, the vaccine composition is for administration to livestock animals. In some embodiments, the vaccine composition is for administration to cows. In some embodiments, the vaccine composition is for administration to sheep. In some embodiments, the vaccine composition is for administration to swine. In some embodiments, the vaccine composition is for administration to avian animals. In some embodiments, the vaccine composition is for administration to canine animals. In some embodiments, the vaccine composition is for administration to feline animals. In some embodiments, the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is from about 1 nanogram (ng) to about 3 micrograms (μg). In some embodiments, the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is from about 1 nanogram (ng) to about 1 microgram (μg). In some embodiments, the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is from about 1 nanogram (ng) to about 0.1 microgram (μg). In some embodiments, the homologous distinct antigenic component comprises a viral component. In some embodiments, the viral component is a receptor-binding domain.In some embodiments, at least two of the six or more homologous different antigenic components comprise receptor binding domains for a cell surface protein. In some embodiments, the cell surface protein is mammalian. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 99% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 98% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 95% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous different antigenic components share less than 90% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 85% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 80% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 75% sequence identity. In some embodiments, the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 70% sequence identity. In some embodiments, administration of the vaccine composition reduces the predominance of immunogenic single variant epitopes corresponding to the homologous distinct antigenic components. In some embodiments, any two of the six or more homologous distinct antigenic components share less than about 90% sequence identity.In some embodiments, any two of the six or more homologous different antigenic components share less than about 85% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 80% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 75% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share less than about 70% sequence identity. In some embodiments, the vaccine composition comprises seven or more homologous different antigenic components. In some embodiments, the vaccine composition comprises ten or more homologous different antigenic components. In some embodiments, the vaccine composition comprises fifteen or more homologous different antigenic components. In some embodiments, the vaccine composition comprises twenty or more homologous different antigenic components. In some embodiments, the vaccine composition comprises twenty-five or more homologous different antigenic components. In some embodiments, the vaccine composition comprises thirty or more homologous different antigenic components. In some embodiments, any two of the six or more homologous different antigenic components share at least about 25% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 30% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 35% sequence identity. In some embodiments, the vaccine composition further comprises one or more non-homologous antigenic components. In some embodiments, the vaccine composition further comprises an adjuvant. In some embodiments, the six or more homologous different antigenic components are present in the vaccine composition at the same concentration. In some embodiments, the six or more homologous different antigenic components are present in the vaccine composition at different concentrations / amounts.In some embodiments, a dose of a homologous distinct antigenic component among six or more homologous distinct antigenic components is calculated to be proportional to the phylogenetic distance of the homologous distinct antigenic component to another homologous distinct antigenic component, where another homologous distinct antigenic component has the smallest distance to the homologous distinct antigenic component among the six or more homologous distinct antigenic components. In some embodiments, a dose of a homologous distinct antigenic component among six or more homologous distinct antigenic components is calculated to be proportional to its average phylogenetic distance from other homologous distinct antigenic components among the six or more homologous distinct antigenic components. In some embodiments, the vaccine composition comprises a fragment of a SARS virus. In some embodiments, the vaccine composition comprises a fragment of an influenza virus. In some embodiments, the vaccine composition comprises a fragment of an HIV virus. In some embodiments, the vaccine composition comprises a fragment of a SARS1 virus.

[0009] Another aspect of the present disclosure provides a method of inducing an immune response against a pathogen in a subject, the method comprising administering a vaccine composition disclosed herein. In some embodiments, the method is for prophylaxis against a disease caused by a pathogen. Further aspects and advantages of the present disclosure will become readily apparent to those skilled in the art from the following detailed description, wherein only exemplary embodiments of the present disclosure are shown and described. As will be understood, the present disclosure is capable of other and different embodiments, and its several details are capable of modification in various obvious respects, all without departing from the present disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature and not restrictive.

[0010] BRIEF DESCRIPTION OF THE DRAWINGS The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings (also referred to herein as "Figure" and "FIG."). [Brief explanation of the drawings]

[0011] [Figure 1] Figure 1 shows the number of FDA approved vaccines for various doses (μg) per component of the vaccine. There are no FDA approved vaccines with a dose per component less than 3.75 μg.

[0012] [Figure 2A] Figure 2A shows the amount of induced antibodies, measured by ELISA, reactive to specific recombinant influenza hemagglutinin antigens in the serum of mice in response to vaccination with either Centi-Flu (0.031 μg mRNA / antigen), Flu-Biv (0.25 μg / antigen), or Flu-Biv (0.031 μg / antigen). Reactivity is demonstrated with antigens present in Centi-Flu (H3N2 Hong Kong / 1 / 1968, H3N2 Alaska / 01 / 2021), antigens present in both Centi-Flu and Flu-Biv (California / 07 / 2004), or heterologous antigens not present in either (H3N2A / Victoria / 361 / 2011, A / Maryland / 02 / 2021). Figure 2B shows the amount of induced antibodies, measured by ELISA, reactive to specific recombinant influenza hemagglutinin antigens in the serum of mice in response to vaccination with either Centi-Flu (0.25 μg mRNA / antigen), Flu-Biv (2 μg / antigen), or Flu-Biv (0.25 μg / antigen). Reactivity is demonstrated with antigens present in Centi-Flu (H3N2 Hong Kong / 1 / 1968, H3N2 Alaska / 01 / 2021), antigens present in both Centi-Flu and Flu-Biv (California / 07 / 2004), or heterologous antigens not present in either (H3N2A / Victoria / 361 / 2011, A / Maryland / 02 / 2021). [Figure 2B] Same as above.

[0013] [Figure 3A]Figure 3A shows seroreactivity measured by ELISA in ferrets immunized with 1 μg of mRNA encoding influenza hemagglutinin antigens for H3N2 Alaska / 01 / 2021, H3N2 California / 07 / 2004, H3N2 Cambodia / 2020, H3N2 Indiana / 11 / 2018, H3N2 Bilthoven / 1761 / 1976, H3N2 Indiana / 08 / 2011, H3N2 Nanchang / 933 / 1995, or H3N2 Memphis / 1 / 1980 against the same antigens with which the animals were immunized, and Figure 3B shows the seroreactivity measured by ELISA in ferrets immunized with 1 μg of mRNA encoding influenza hemagglutinin antigens for H3N2 Alaska / 01 / 2021, H3N2 California / 07 / 2004, H3N2 Cambodia / 2020, H3N2 Indiana / 11 / 2018, H3N2 Bilthoven / 1761 / 1976, H3N2 Indiana / 08 / 2011, H3N2 Nanchang / 933 / 1995, or H3N2 Memphis / 1 / 1980 against the same antigens with which the animals were immunized. Figure 1 shows seroreactivity measured by ELISA in ferrets immunized with approximately 0.5 μg of mRNA per antigen encoding eight influenza hemagglutinin antigens: H3N2 Alaska / 01 / 2021, H3N2 California / 07 / 2004, H3N2 Cambodia / 2020, H3N2 Indiana / 11 / 2018, H3N2 Bilthoven / 1761 / 1976, H3N2 Nanchang / 933 / 1995, H3N2 Memphis / 1 / 1980, and H3N2 Hong Kong / 1 / 1968, to the indicated recombinant protein antigens. [Figure 3B] Same as above.

[0014] [Figure 4] Figure 4 shows seroreactivity measured by ELISA in mice immunized with either Centi-HIV (10 recombinant gp160 antigens; 1 ug / human equivalent dose (HED) of antigen = 10 ug total), or single recombinant HIV gp160 antigens (HED 1 ug or 10 ug).

[0015] [Figure 5] Figure 5 shows seroreactivity measured by ELISA against snake venom from Crotalus horridus in mice immunized with either a mixture of 15 snake venoms (3ug / ag effective HED = 45ug total) or Crotalus horridus snake venom alone (3ug effective HED or 45ug).

[0016] [Figure 6]FIG. 6 shows the pairwise sequence identity between a set of different antigenic components homologous to the H3N2 hemagglutinin.

[0017] [Figure 7] FIG. 7 shows the pairwise sequence identity between a set of different antigenic components homologous to HIV gp160.

[0018] [Figure 8] FIG. 8 shows pairwise sequence identities between different antigenic components homologous to a set of snake venom PLA2s. DETAILED DESCRIPTION OF THE INVENTION

[0019] Detailed Description While various embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions may be made by those skilled in the art without departing from the invention. It is understood that various alternatives to the embodiments of the invention described herein may be used.

[0020] Vaccines may contain multiple homologous antigenic components, particularly for pathogens belonging to diverse phylogenetic families (e.g., influenza). For any particular antigenic component, there may be a minimal dose of that antigenic component sufficient to elicit an optimal immune response to that antigen alone. The present disclosure describes vaccine compositions comprising a mixture of multiple homologous antigenic components, each of which is administered below such a threshold.

[0021] In some embodiments, the vaccine composition is an RNA vaccine. For RNA vaccines, a typical dose per antigen component can be at least about 30 μg to at least about 100 μg. Examples of such RNA vaccines can include the Pfizer or Moderna COVID vaccines. Dose-escalation studies have shown that about 10 μg may be a threshold dose in adults that is insufficient to elicit a sufficient immune response. The RNA can be messenger RNA (mRNA). The mRNA can encode an antigenic protein. The antigen can be an influenza protein or a fragment thereof. The antigen can be a hemagglutinin protein or a fragment thereof. In the context of the RNA vaccines disclosed herein, "antigenic component" can refer to the mRNA encoding the protein component against which the vaccine is directed. In the context of the RNA vaccines disclosed herein, sequence identity between two antigenic components can refer to sequence identity between the amino acid sequences of the protein components encoded by the mRNA.

[0022] In the context of the non-RNA vaccines disclosed herein, "antigenic component" can refer to the protein component against which the antigen is directed.

[0023] In some embodiments, the vaccine composition is a virus-like particle (VLP) vaccine. For VLP vaccines, typical doses per antigen component are at least about 20 μg to at least about 40 μg. The minimum reported dose may be about 5 μg for infants, children, and adolescents. The minimum reported dose may be about 10 μg for adults. Examples of minimum dose vaccines of about 5 μg for infants, children, and adolescents include RECOMBIVAX HB. Examples of minimum dose vaccines of about 10 μg for adults include RECOMBIVAX HB.

[0024] The minimum per-antigen dose of any clinically approved vaccine in a particular format means that administration below that dose will not elicit a protective immune response, or the elicited immune response will be suboptimal compared to higher doses. The present disclosure provides a novel vaccine approach in which five or more antigens are combined into a single vaccine, with each individual antigen dose being below the minimum dose utilized in any clinically approved vaccine of that format. This may enable the selective stimulation of B cells that recognize broadly conserved epitopes by combining the preservation of these epitopes with their concentration / amount in the vaccine formulation. By administering a mixture of many (i.e., ≥ 5) diverse variants of a recombinant antigen, each individually, below the threshold dose required to elicit an optimal strain-specific immune response, the vaccine compositions disclosed herein may reduce the predominance of immunogenic single variant epitopes, leaving conserved epitopes (found across multiple components) at higher effective doses. Conserved epitopes may be primarily represented in the vaccine compositions claimed herein, even though each immunogen retains its intact native structure. Furthermore, the conserved conformational epitopes are completely conserved.

[0025] In some embodiments, two antigenic components are considered "different" if their amino acid sequences, or their corresponding amino acid sequences (in the case of RNA vaccines), share less than about 99%, 98%, 97%, 96%, 95%, 90%, 85%, 80%, 75%, or 70% sequence identity.

[0026] In other embodiments, two antigen components are considered "different" if they comprise receptor-binding domains of mammalian cell surface proteins and the amino acid sequences of their respective receptor-binding domains share less than about 99%, 98%, 96%, 95%, 90%, 85%, 80%, 75%, or 70% sequence identity.

[0027] In some embodiments, the vaccine compositions of the invention comprise six or more homologous different antigenic components. In some embodiments, any two of the six or more homologous different antigenic components share at least about 30% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 40% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 50% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 60% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 70% sequence identity. In some embodiments, any two of the six or more homologous different antigenic components share at least about 80% sequence identity.

[0028] In some embodiments, the vaccine compositions of the present invention comprise six or more RNAs encoding homologous, distinct antigenic components. In some embodiments, each RNA is present in an amount of at least 10 nanograms per dose. In some embodiments, each RNA is present in an amount of at least 100 nanograms per dose. In some embodiments, each RNA is present in an amount of at least 250 nanograms per dose. In some embodiments, each RNA is present in an amount of at least 500 nanograms per dose.

[0029] The vaccine compositions of the present invention comprise five or more homologous antigen components, each pair being different ("distinct sets"). The distinct sets may comprise approximately 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more than 30 homologous antigen components. The vaccine compositions may further comprise additional homologous antigen components that are not different for the distinct sets or that are different from each other. For example, a vaccine composition of the present invention may comprise eight antigen components. Components 1-6 form distinct sets in which any two share less than 95% sequence identity and components 7-8 share more than 95% sequence identity with each other. As another example, a vaccine composition of the present invention may comprise eight antigen components. Components 1-6 form a distinct set where any two share less than 95% sequence identity and components 7-8 share more than 95% sequence identity to component 1. The vaccine composition may further comprise additional heterologous antigen components.

[0030] In some embodiments, the dose of each antigen component in a protein vaccine intended for adult use is more than about 1 ng per dose but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose. In some embodiments, the dose of each antigen component in a protein vaccine intended for adult use is more than about 600 ng per dose, but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, or less than about 1 μg per dose.

[0031] In some embodiments, the dose of each antigen component in a protein vaccine intended for pediatric use is more than about 1 ng per dose but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose. In some embodiments, the dose of each antigen component in a protein vaccine intended for pediatric use is more than about 600 ng per dose, but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, or less than about 1 μg per dose.

[0032] In some embodiments, the dose of each antigen component in a protein vaccine intended for veterinary use is more than about 1 ng per dose but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose.

[0033] In some embodiments, the dose of each antigen component in an RNA vaccine intended for adult use is greater than about 1 ng per dose but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose. In some embodiments, the dose of each antigen component in an RNA vaccine intended for adult use is greater than about 10 ng per dose but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose. In some embodiments, the dose of each antigen component in an RNA vaccine intended for adult use is greater than about 100 ng per dose, but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, or less than about 0.5 μg per dose.

[0034] In some embodiments, the dose of each antigen component in an RNA vaccine intended for pediatric use is greater than about 1 ng per dose but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose. In some embodiments, the dose of each antigen component in an RNA vaccine intended for pediatric use is more than about 10 ng per dose but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose. In some embodiments, the dose of each antigen component in an RNA vaccine intended for pediatric use is more than about 100 ng per dose, but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, or less than about 0.5 μg per dose.

[0035] In some embodiments, the dose of each antigen component in an RNA vaccine intended for veterinary use is greater than about 1 ng per dose but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose. In some embodiments, the dose of each antigen component in an RNA vaccine intended for veterinary use is greater than about 10 ng per dose, but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose. In some embodiments, the dose of each antigen component in an RNA vaccine intended for veterinary use is more than about 100 ng per dose, but less than about 10 μg per dose, less than about 9 μg per dose, less than about 8 μg per dose, less than about 7 μg per dose, less than about 6 μg per dose, less than about 5 μg per dose, less than about 4 μg per dose, less than about 3 μg per dose, less than about 2 μg per dose, less than about 1 μg per dose, or less than about 0.5 μg per dose.

[0036] In some embodiments, vaccine compositions of the invention comprise RNA encoding six or more homologous, distinct antigenic components, where each pair of antigens shares 25%-96% sequence identity, and the amount of each RNA is 50 ng per dose to 2 μg per dose. In some embodiments, human adult vaccine compositions of the invention comprise RNA encoding six or more homologous, distinct antigenic components, where each pair of antigens shares 25%-96% sequence identity, and the amount of each RNA is 50 ng per dose to 5 μg per dose. In some embodiments, human pediatric vaccine compositions of the invention comprise RNA encoding six or more homologous, distinct antigenic components, where each pair of antigens shares 25%-96% sequence identity, and the amount of each RNA is 10 ng per dose to 2 μg per dose.

[0037] In some embodiments, the vaccine compositions of the invention comprise six or more homologous distinct antigenic components, where each pair of antigens shares between 25% and 96% sequence identity, and the amount of each antigenic component is between 600 ng per dose and 3 μg per dose.

[0038] In some embodiments, the antigenic component can be an inactivated virus, in which case the dose for a given protein of antigenic interest is calculated as follows: Dose of a given protein of antigenic target = (estimated number of viral particles in the dose) x (number of copies of the protein of antigenic target per viral particle) x (molecular weight of the protein of antigenic target).

[0039] In some embodiments, the vaccine composition may further comprise an adjuvant.

[0040] In some embodiments, each antigen component of the vaccine composition may be present in the same dose as the other antigen components.

[0041] In other embodiments, each antigenic component of the vaccine composition may be present in a different dose than the other antigenic components. The dose of a given antigenic component may be calculated to be proportional to that component's phylogenetic distance to the nearest other component, or may be calculated to be proportional to its average phylogenetic distance to all other components.

[0042] For protein subunit vaccines, a typical dose per antigen component may be at least about 15 μg to at least about 30 μg for adults. For protein subunit vaccines, a typical dose per antigen component may be at least about 7.5 μg to at least about 15 μg for children. The minimum dose of a clinically approved protein subunit vaccine may be at least about 3.75 μg. An example of such a clinically approved protein subunit vaccine may include the monovalent H5N1 influenza HA vaccine (ID Biomedical Corporation).

[0043] In some embodiments, the invention includes human adult vaccine compositions comprising six or more homologous distinct antigenic components, where each pair of antigens shares less than about 95% sequence identity, and the antigenic components are proteins and each distinct antigenic component is less than about 3 μg per dose, or where the antigenic components are RNA and each distinct antigenic component is less than about 5 μg per dose, or where the antigenic components are proteins displayed on heterologous VLPs and each distinct antigenic component is less than about 5 μg per dose. In some embodiments, the invention includes human adult vaccine compositions comprising six or more homologous distinct antigenic components, where each pair of antigens shares less than about 95% sequence identity, and the antigenic components are proteins and each distinct antigenic component is less than about 3 μg per dose to more than about 600 ng per dose. In some embodiments, the invention includes human adult vaccine compositions comprising mRNA encoding six or more homologous distinct antigenic components, where each pair of antigenic components shares less than about 95% sequence identity, and each distinct antigenic component is less than about 5 μg per dose to more than about 50 ng per dose.

[0044] In some embodiments, the invention includes human pediatric vaccine compositions comprising six or more homologous distinct antigenic components, each pair of antigens sharing less than about 95% sequence identity, and the antigenic components are proteins and each distinct antigenic component is less than about 1 μg per dose, or the antigenic components are RNA and each distinct antigenic component is less than about 2.5 μg per dose, or the antigenic components are proteins displayed on heterologous VLPs and each distinct antigenic component is less than about 2.5 μg per dose. In some embodiments, the invention includes human pediatric vaccine compositions comprising mRNA encoding six or more homologous distinct antigenic components, each pair of antigenic components sharing less than about 95% sequence identity, and the mRNA encoding each distinct antigenic component is less than about 1 μg per dose to more than about 10 ng per dose.

[0045] In some embodiments, the invention includes veterinary vaccine compositions comprising six or more homologous different antigenic components, wherein each pair of antigens shares less than about 95% sequence identity, and wherein the antigenic components are proteins and each different antigenic component is less than about 3 μg per dose, or wherein the antigenic components are RNA and each different antigenic component is less than about 5 μg per dose, or wherein the antigenic components are proteins displayed on heterologous VLPs and each different antigenic component is less than about 5 μg per dose.

[0046] In some embodiments, the vaccine compositions of the present invention comprise RNA encoding homologous distinct antigenic components. In some embodiments, the RNA is messenger RNA (mRNA). In some embodiments, the RNA is encapsulated in a lipid nanoparticle (LNP). In some embodiments, the LNP comprises an ionizable or cationic lipid, a helper phospholipid, a PEG-conjugated lipid, and a cholesterol-based lipid. In some embodiments, the ionizable lipid is ALC-0315, SM-102, or MC-3. In some embodiments, the cholesterol-based lipid is cholesterol. In some embodiments, the helper phospholipid is DSPC. In some embodiments, the PEG-conjugated lipid is ALC-0159. In some embodiments, the ionizable lipid is present in a molar ratio of 35-55%. In some embodiments, the PEG-conjugated lipid is present in a molar ratio of 0.25-3%. In some embodiments, the helper phospholipid is present in a molar ratio of 5-20%. In some embodiments, the cholesterol-based lipid is present in a molar ratio of 20-45%. All lipid molar ratios described herein are relative to the total lipid content of the LNPs.

[0047] In certain embodiments, the mRNA comprises at least one chemical modification. In certain embodiments, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the uracil nucleotides of the mRNA are chemically modified. In certain embodiments, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the uracil nucleotides of the ORF are chemically modified. In certain embodiments, the chemical modification is selected from the group consisting of pseudouridine, N1-methylpseudouridine, 2-thiouridine, 4'-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine, 5-methyluridine, 5-methoxyuridine, and 2'-O-methyluridine. In certain embodiments, the chemical modification is selected from the group consisting of pseudouridine, N1-methylpseudouridine, 5-methylcytosine, 5-methoxyuridine, and combinations thereof. In certain embodiments, the chemical modification is N1-methylpseudouridine.

[0048] Influenza antigen

[0049] The present disclosure provides a vaccine comprising a plurality of RNAs encoding six or more homologous distinct antigenic components. The vaccine may be for the prevention of influenza. The six or more homologous distinct antigenic components may comprise one or more influenza proteins or one or more fragments thereof. The six or more homologous distinct antigenic components may comprise a hemagglutinin component. In some embodiments, one or more hemagglutinin components are derived from the H1N1 subtype. In some embodiments, one or more hemagglutinin components are derived from the H3N2 subtype.

[0050] In some embodiments, the invention includes human adult vaccine compositions comprising a plurality of RNAs encoding six or more homologous distinct antigenic components, wherein the homologous distinct antigenic components are influenza hemagglutinin or fragments thereof, each pair of antigenic components sharing less than about 96% sequence identity, and the amount of each mRNA is less than about 5 μg per dose. In some embodiments, the invention includes vaccine compositions comprising a plurality of RNAs encoding six or more homologous distinct antigenic components, wherein the homologous distinct antigenic components are influenza hemagglutinin or fragments thereof, each pair of antigenic components sharing less than about 96% sequence identity, and the amount of each mRNA is less than about 4 μg per dose, less than about 3 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose. In some embodiments, the invention includes vaccine compositions comprising a plurality of RNAs encoding six or more homologous distinct antigenic components, wherein the homologous distinct antigenic components are influenza hemagglutinin or fragments thereof, and each pair of antigenic components shares less than about 95% sequence identity, and the amount of each mRNA is less than about 4 μg per dose, less than about 3 μg per dose, less than about 1 μg per dose, less than about 0.5 μg per dose, or less than about 0.1 μg per dose.

[0051] In some embodiments, vaccine compositions of the invention comprise RNA encoding six or more homologous distinct antigenic components, where the homologous distinct antigenic components are influenza hemagglutinin or fragments thereof. In some embodiments, each pair of antigens shares 25% to 96% sequence identity, and the amount of each RNA is 50 ng per dose to 2 μg per dose. In some embodiments, the amount of each RNA is 50 ng per dose to 5 μg per dose. In some embodiments intended for pediatric use, the amount of each RNA is 10 ng per dose to 2 μg per dose.

[0052] The present disclosure provides vaccines for preventing influenza. In some embodiments, one or more hemagglutinin components are derived from one or more influenza strains. In some embodiments, the one or more influenza strains include A / Wisconsin / 588 / 2019. In some embodiments, the one or more influenza strains include A / California / 07 / 2009. In some embodiments, the one or more influenza strains include A / Denver / 57. In some embodiments, the one or more influenza strains include A / Brisbane / 59 / 2007. In some embodiments, the one or more influenza strains include A / Pekin / 262 / 1995. In some embodiments, the one or more influenza strains include A / Puerto Rico / 8 / 1934. In some embodiments, the one or more influenza strains include A / New York / 1 / 1918. In some embodiments, the one or more influenza strains include A / Wisconsin / 28 / 2011, 2011 / 12. In some embodiments, the one or more influenza strains include A / Indiana / 11 / 2018. In some embodiments, the one or more influenza strains include A / California / 07 / 2004. In some embodiments, the one or more influenza strains include A / Alaska / 01 / 2021.

[0053] In some embodiments, one or more influenza strains comprise A / Cambodia / e0826360 / 2020. In some embodiments, one or more influenza strains comprise A / Nanchang / 933 / 1995. In some embodiments, one or more influenza strains comprise A / Memphis / 1 / 1980. In some embodiments, one or more influenza strains comprise A / Bilthoven / 1761 / 1976. In some embodiments, one or more influenza strains comprise A / Hong Kong / 1 / 1968. In some embodiments, one or more influenza strains comprise A / Indiana / 08 / 2011. In some embodiments, the antigen included in or encoded by the vaccine has at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95% or about 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33 or 35.

[0054] The present disclosure provides a vaccine for preventing influenza, the vaccine comprising a plurality of RNAs encoding a plurality of different homologous antigenic components. The different homologous antigenic components can be influenza proteins or fragments thereof. In some embodiments, one RNA of the plurality of RNAs is messenger RNA (mRNA). In some embodiments, the mRNA encodes an influenza antigen or fragment thereof. In some embodiments, the mRNA encodes an influenza hemagglutinin or fragment thereof. In some embodiments, the mRNA encoding the influenza antigen comprises an mRNA insert region. In some embodiments, the mRNA insertion region has at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% identity to any one of SEQ ID NOs: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36, and a thymine in any one of SEQ ID NOs: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36 is replaced with a uridine, a uridine isomer, pseudouridine, a pseudouridine methylated derivative, or N1-methyl-pseudouridine. In some embodiments, the mRNA encoding the influenza antigen comprises a 3'UTR region. In some embodiments, the 3'UTR region has at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% identity to SEQ ID NO: 2, wherein the thymine in SEQ ID NO: 2 is replaced with a uridine, a uridine isomer, pseudouridine, a pseudouridine methylated derivative, or N1-methyl-pseudouridine. In some embodiments, the mRNA encoding the influenza antigen comprises a 5'UTR region.In some embodiments, the 5'UTR comprises at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% identity to SEQ ID NO:1, wherein the thymine in SEQ ID NO:1 is replaced with a uridine, a uridine isomer, pseudouridine, a pseudouridine methylated derivative, or N1-methyl-pseudouridine. Vaccine sequence list [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] [Table 1-13] [Table 1-14] Table 1-15 Table 1-16 Table 1-17 Table 1-18 Table 1-19 Table 1-20 Table 1-21 Table 1-22 Table 1-23 Table 1-24 Table 1-25 Table 1-26 Table 1-27 Table 1-28 Table 1-29 Table 1-30 Table 1-31 Table 1-32 Table 1-33 Table 1-34 Table 1-35 Table 1-36 Table 1-37 Table 1-38 Table 1-39 Table 1-40 Table 1-41 Table 1-42 Table 1-43 Table 1-44 Table 1-45 Table 1-46 Table 1-47 Table 1-48 [Table 1-49] Numbering implementation example Embodiment 1. A vaccine composition comprising six or more homologous distinct antigenic components, wherein any two of the six or more homologous distinct antigenic components share less than 95% sequence identity; a. the homologous distinct antigenic component comprises a protein, and the concentration / amount of the protein in a dose of a human adult vaccine composition is from about 1 nanogram (ng) to about 3 micrograms (μg); b. the homologous distinct antigenic components comprise a plurality of RNAs, and the concentration / amount of RNA of the plurality of RNAs in a dose of the vaccine composition is from about 1 ng to about 5 μg per dose; or c. The homologous distinct antigenic components comprise multiple proteins displayed on heterologous virus-like particles (VLPs), and the concentration / amount of the multiple proteins displayed on the heterologous VLPs in a dose of the vaccine composition is from about 1 ng to about 5 μg per dose. Embodiment 2. The vaccine composition of embodiment 1, wherein the vaccine composition is for administration to a human subject. Embodiment 3. The vaccine composition of embodiment 2, wherein the vaccine composition is for administration to adults 18 years of age or older. Embodiment 4. The vaccine composition of embodiment 3, wherein the vaccine composition is for administration to adults 25 years of age or older. Embodiment 5. The vaccine composition of embodiment 3, wherein the vaccine composition is for administration to adults 50 years of age or older. Embodiment 6 The vaccine composition of embodiment 3, wherein the vaccine composition is for administration to adults 75 years of age or older. Embodiment 7. The vaccine composition of embodiment 2, wherein the vaccine composition is for administration to children aged from about 1 day to about 18 years. Embodiment 8. The vaccine composition of embodiment 2, wherein the vaccine composition is for administration to children aged from about 1 day to about 5 years. Embodiment 9. The vaccine composition of embodiment 2, wherein the vaccine composition is for administration to children between the ages of about 5 years and about 18 years. Embodiment 10. The vaccine composition of embodiment 1, wherein the vaccine composition is for administration to an animal. Embodiment 11 The vaccine composition of embodiment 10, wherein the animal is a livestock animal. Embodiment 12. The vaccine composition of embodiment 11, wherein the livestock animal is a cow, bull, alpaca, llama, sheep, pig, or bird. Embodiment 13 The vaccine composition of embodiment 10, wherein the animal is a domestic animal. Embodiment 14. The vaccine composition of embodiment 13, wherein the domestic animal is a primate. Embodiment 15. The vaccine composition of embodiment 1, wherein the six or more homologous distinct antigenic components comprise antigens selected from the group consisting of viral, bacterial, fungal, prion, plant, or combinations thereof. Embodiment 16. The vaccine composition of embodiment 1, wherein the concentration / amount of one of the homologous distinct antigenic components in one dose of the vaccine composition is from about 1 nanogram (ng) to about 5 micrograms (μg). Embodiment 17. The vaccine composition of embodiment 1, wherein the concentration / amount of one of the homologous distinct antigenic components in a dose of the vaccine composition is from about 1 nanogram (ng) to about 1 microgram (μg). Embodiment 18. A vaccine composition according to any one of embodiments 1 to 17, wherein the homologous distinct antigenic components comprise components of a virus. Embodiment 19 The vaccine composition of embodiment 18, wherein the viral component is a receptor-binding domain. Embodiment 20. The vaccine composition of embodiment 18, wherein at least two of said six or more homologous distinct antigenic components comprise receptor binding domains for cell surface proteins. Embodiment 21 The vaccine composition of embodiment 20, wherein the cell surface protein is mammalian. Embodiment 22. The vaccine composition of embodiment 20, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 99% sequence identity. Embodiment 23. The vaccine composition of embodiment 22, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 98% sequence identity. Embodiment 24. The vaccine composition of embodiment 23, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 95% sequence identity. Embodiment 25. The vaccine composition of embodiment 24, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 95% sequence identity. Embodiment 26. The vaccine composition of embodiment 25, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 90% sequence identity. Embodiment 27. The vaccine composition of embodiment 26, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 85% sequence identity. Embodiment 28. The vaccine composition of embodiment 27, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 80% sequence identity. Embodiment 29. The vaccine composition of embodiment 28, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 75% sequence identity. Embodiment 30. The vaccine composition of embodiment 29, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 70% sequence identity. Embodiment 31. The vaccine composition of any one of embodiments 1 to 30, wherein administration of the vaccine composition reduces the predominance of immunogenic single mutant epitopes corresponding to the homologous distinct antigenic components. Embodiment 32. The vaccine composition of any one of embodiments 1 to 31, wherein any two of the six or more homologous different antigenic components share less than about 90% sequence identity. Embodiment 33. The vaccine composition of embodiment 31, wherein any two of said six or more homologous different antigenic components share less than about 85% sequence identity. Embodiment 34. The vaccine composition of embodiment 33, wherein any two of said six or more homologous different antigen components share less than about 80% sequence identity. Embodiment 35. The vaccine composition of embodiment 34, wherein any two of said six or more homologous different antigenic components share less than about 75% sequence identity. Embodiment 36. The vaccine composition of embodiment 35, wherein any two of said six or more homologous different antigenic components share less than about 70% sequence identity. Embodiment 37. The vaccine composition of any one of embodiments 1 to 36, wherein the vaccine composition comprises seven or more homologous different antigenic components. Embodiment 38. The vaccine composition of any one of embodiments 1 to 37, wherein the vaccine composition comprises 10 or more homologous different antigenic components. Embodiment 39. The vaccine composition of any one of embodiments 1 to 38, wherein the vaccine composition comprises 15 or more homologous different antigenic components. Embodiment 40. The vaccine composition of any one of embodiments 1 to 39, wherein the vaccine composition comprises 20 or more homologous different antigenic components. Embodiment 41. The vaccine composition of any one of embodiments 1 to 40, wherein the vaccine composition comprises 25 or more homologous different antigenic components. Embodiment 42. The vaccine composition of any one of embodiments 1 to 41, wherein the vaccine composition comprises 30 or more homologous different antigenic components. Embodiment 43. The vaccine composition of any one of embodiments 1 to 42, wherein any three of the six or more homologous different antigenic components share less than 95% sequence identity. Embodiment 44. The vaccine composition of any one of embodiments 1 to 43, wherein any four of the six or more homologous different antigenic components share less than 95% sequence identity. Embodiment 45. The vaccine composition of any one of embodiments 1 to 44, wherein any five of the six or more homologous different antigenic components share less than 95% sequence identity. Embodiment 46. The vaccine composition of any one of embodiments 1 to 45, wherein any six of the six or more homologous different antigen components share less than 95% sequence identity. Embodiment 47. The vaccine composition of any one of embodiments 1 to 46, further comprising one or more heterologous antigen components. Embodiment 48. A vaccine composition according to any one of embodiments 1 to 47, further comprising an adjuvant. Embodiment 49. The vaccine composition of any one of embodiments 1 to 48, wherein the six or more homologous different antigenic components are present in the vaccine composition at the same concentration / amount. Embodiment 50. The vaccine composition of any one of embodiments 1 to 49, wherein the six or more homologous different antigenic components are present in the vaccine composition at different concentrations / amounts. Embodiment 51. A vaccine composition according to any one of embodiments 1 to 50, wherein one dose of a homologous distinct antigenic component among the six or more homologous distinct antigenic components is calculated to be proportional to the phylogenetic distance between the homologous distinct antigenic component and another homologous distinct antigenic component, wherein the other homologous distinct antigenic component has the smallest distance to the homologous distinct antigenic component among the six or more homologous distinct antigenic components. Embodiment 52. A vaccine composition according to any one of embodiments 1 to 51, wherein a dose of a homologous distinct antigenic component among the six or more homologous distinct antigenic components is calculated to be proportional to its average phylogenetic distance from other homologous distinct antigenic components among the six or more homologous distinct antigenic components. Embodiment 53. The vaccine composition of any one of embodiments 1 to 52, wherein the vaccine composition comprises a fragment of a SARS virus. Embodiment 54. A vaccine composition according to any one of embodiments 1 to 53, comprising a fragment of an influenza virus. Embodiment 55. A vaccine composition according to any one of embodiments 1 to 54, wherein the vaccine composition comprises a fragment of the HIV virus. Embodiment 56. The vaccine composition of any one of embodiments 1 to 55, wherein the vaccine composition comprises a fragment of the SARS1 virus. Embodiment 57. A vaccine composition for human adults comprising six or more homologous distinct antigenic components, wherein two of the six or more homologous distinct antigenic components share less than 95% sequence identity; a. the homologous distinct antigenic component comprises a protein, and the concentration / amount of protein in a dose of the human adult vaccine composition is from about 1 nanogram (ng) to about 3 micrograms (μg); b. the homologous distinct antigenic components comprise a plurality of RNAs, and the concentration / amount of RNA of the plurality of RNAs in a dose of the human adult vaccine composition is from about 1 ng to about 5 μg per dose; or c. The homologous distinct antigenic components comprise multiple proteins presented on heterologous virus-like particles (VLPs), and the concentration / amount of the multiple proteins presented on heterologous VLPs in a dose of the human adult vaccine composition is from about 1 ng to about 5 μg per dose. Embodiment 58 The vaccine composition of embodiment 57, wherein the vaccine composition is for administration to adults 25 years of age or older. Embodiment 59. The vaccine composition of embodiment 57, wherein the vaccine composition is for administration to adults 50 years of age or older. Embodiment 60 The vaccine composition of embodiment 57, wherein the vaccine composition is for administration to adults 75 years of age or older. Embodiment 61. The vaccine composition of any one of embodiments 57 to 60, wherein the concentration / amount of the homologous distinct antigenic component in one dose of the vaccine composition is from about 1 nanogram (ng) to about 2 μg. Embodiment 62. The vaccine composition of embodiment 61, wherein the concentration / amount of one of the homologous distinct antigenic components in one dose of the vaccine composition is from about 1 nanogram (ng) to about 1 microgram (μg). Embodiment 63. The vaccine composition of embodiment 62, wherein the concentration / amount of one of the homologous distinct antigenic components in one dose of the vaccine composition is from about 1 nanogram (ng) to about 0.1 microgram (μg). Embodiment 64. A vaccine composition according to any one of embodiments 57 to 63, wherein the homologous distinct antigenic components comprise components of a virus. Embodiment 65. The vaccine composition of embodiment 64, wherein the viral component is a receptor-binding domain. Embodiment 66. The vaccine composition of embodiment 64, wherein at least two of the six or more homologous distinct antigenic components comprise receptor-binding domains for cell surface proteins. Embodiment 67 The vaccine composition of embodiment 66, wherein the cell surface protein is mammalian. Embodiment 68. The vaccine composition of embodiment 66, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 99% sequence identity. Embodiment 69. The vaccine composition of embodiment 68, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 98% sequence identity. Embodiment 70. The vaccine composition of embodiment 69, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 95% sequence identity. Embodiment 71. The vaccine composition of embodiment 70, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 95% sequence identity. Embodiment 72. The vaccine composition of embodiment 71, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 90% sequence identity. Embodiment 73. The vaccine composition of embodiment 72, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 85% sequence identity. Embodiment 74. The vaccine composition of embodiment 73, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 80% sequence identity. Embodiment 75. The vaccine composition of embodiment 74, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 75% sequence identity. Embodiment 76. The vaccine composition of embodiment 75, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 70% sequence identity. Embodiment 77. A vaccine composition according to any one of embodiments 57 to 76, wherein administration of the vaccine composition reduces the predominance of immunogenic single mutant epitopes corresponding to the homologous distinct antigenic components. Embodiment 78. The vaccine composition of any one of embodiments 57 to 77, wherein any two of the six or more homologous different antigen components share less than about 90% sequence identity. Embodiment 79. The vaccine composition of embodiment 78, wherein any two of said six or more homologous different antigenic components share less than about 85% sequence identity. Embodiment 80. The vaccine composition of embodiment 79, wherein any two of said six or more homologous different antigen components share less than about 80% sequence identity. Embodiment 81. The vaccine composition of embodiment 80, wherein any two of said six or more homologous different antigenic components share less than about 75% sequence identity. Embodiment 82. The vaccine composition of embodiment 81, wherein any two of said six or more homologous different antigen components share less than about 70% sequence identity. Embodiment 83. The vaccine composition of any one of embodiments 57 to 82, wherein the vaccine composition comprises seven or more homologous different antigenic components. Embodiment 84. The vaccine composition of any one of embodiments 57 to 83, wherein the vaccine composition comprises 10 or more homologous different antigen components. Embodiment 85. The vaccine composition of any one of embodiments 57 to 84, wherein the vaccine composition comprises 15 or more homologous different antigen components. Embodiment 86. The vaccine composition of any one of embodiments 57 to 85, wherein the vaccine composition comprises 20 or more homologous different antigenic components. Embodiment 87. The vaccine composition of any one of embodiments 57 to 86, wherein the vaccine composition comprises 25 or more homologous different antigen components. Embodiment 88. The vaccine composition of any one of embodiments 57 to 87, wherein the vaccine composition comprises 30 or more homologous different antigen components. Embodiment 89. The vaccine composition of any one of embodiments 57 to 87, wherein any three of the six or more homologous different antigen components share less than 95% sequence identity. Embodiment 90. The vaccine composition of any one of embodiments 57 to 87, wherein any four of the six or more homologous different antigenic components share less than 95% sequence identity. Embodiment 91. The vaccine composition of any one of embodiments 57 to 87, wherein any five of the six or more homologous different antigenic components share less than 95% sequence identity. Embodiment 92. The vaccine composition of any one of embodiments 57 to 87, wherein any six of the six or more homologous different antigen components share less than 95% sequence identity. Embodiment 93. The vaccine composition of any one of embodiments 57 to 87, wherein any two of the six or more homologous different antigen components share at least about 95% sequence identity. Embodiment 94. The vaccine composition of any one of embodiments 57 to 87, wherein any three of the six or more homologous different antigenic components share at least about 95% sequence identity. Embodiment 95. The vaccine composition of any one of embodiments 57 to 94, wherein any four of the six or more homologous different antigenic components share at least about 95% sequence identity. Embodiment 96. The vaccine composition of any one of embodiments 57 to 95, further comprising one or more heterologous antigen components. Embodiment 97. A vaccine composition according to any one of embodiments 57 to 96, further comprising an adjuvant. Embodiment 98. The vaccine composition of any one of embodiments 57 to 97, wherein the six or more homologous different antigenic components are present in the vaccine composition at the same concentration / amount. Embodiment 99. The vaccine composition of any one of embodiments 57 to 98, wherein the six or more homologous different antigenic components are present in the vaccine composition at different concentrations / amounts. Embodiment 100. The vaccine composition of any one of embodiments 57 to 99, wherein a dose of a homologous distinct antigenic component among the six or more homologous distinct antigenic components is calculated to be proportional to the phylogenetic distance between the homologous distinct antigenic component and another homologous distinct antigenic component, wherein the other homologous distinct antigenic component has the smallest distance to the homologous distinct antigenic component among the six or more homologous distinct antigenic components. Embodiment 101. The vaccine composition of any one of embodiments 57 to 100, wherein a dose of a homologous distinct antigenic component among the six or more homologous distinct antigenic components is calculated to be proportional to its average phylogenetic distance from other homologous distinct antigenic components among the six or more homologous distinct antigenic components. Embodiment 102. The vaccine composition of any one of embodiments 57 to 101, wherein the vaccine composition comprises a fragment of a SARS virus. Embodiment 103. A vaccine composition according to any one of embodiments 57 to 102, comprising a fragment of an influenza virus. Embodiment 104. The vaccine composition of any one of embodiments 57 to 103, wherein the vaccine composition comprises a fragment of the HIV virus. Embodiment 105. The vaccine composition of any one of embodiments 57 to 104, wherein the vaccine composition comprises a fragment of the SARS1 virus. Embodiment 106. A vaccine composition comprising six or more homologous different antigenic components, wherein two of the six or more homologous different antigenic components share less than 95% sequence identity; a. the homologous distinct antigenic component comprises a protein, and the concentration / amount of protein in a dose of the vaccine composition is from about 1 nanogram (ng) to about 1 microgram (μg); b. the homologous distinct antigenic components comprise a plurality of RNAs, and the concentration / amount of RNA of the plurality of RNAs in a dose of the vaccine composition is from about 1 ng to about 2.5 μg per dose; or c. The homologous distinct antigenic components comprise a plurality of proteins presented on heterologous virus-like particles (VLPs), and the concentration / amount of the plurality of proteins presented on the heterologous VLPs in a dose of the vaccine composition is from about 1 ng to about 2.5 μg per dose, and the vaccine composition is a human pediatric vaccine composition. Embodiment 107. The vaccine composition of embodiment 106, wherein the vaccine composition is for administration to children aged from about 1 day to about 18 years. Embodiment 108. The vaccine composition of embodiment 106, wherein the vaccine composition is for administration to children aged from about 1 day to about 5 years. Embodiment 109. The vaccine composition of embodiment 106, wherein the vaccine composition is for administration to children aged from about 5 years to about 18 years. Embodiment 110. The vaccine composition of any one of embodiments 106 to 109, wherein the concentration / amount of the homologous distinct antigenic component of the homologous distinct antigenic component in a dose of the vaccine composition is from about 1 nanogram (ng) to about 0.5 micrograms (μg). Embodiment 111. The vaccine composition of embodiment 106, wherein the concentration / amount of one of the homologous distinct antigenic components in one dose of the vaccine composition is from about 1 nanogram (ng) to about 0.1 microgram (μg). Embodiment 112. A vaccine composition according to any one of embodiments 106 to 111, wherein the homologous distinct antigenic components comprise components of a virus. Embodiment 113. The vaccine composition of embodiment 112, wherein the viral component is a receptor-binding domain. Embodiment 114. The vaccine composition of embodiment 113, wherein at least two of the six or more homologous distinct antigenic components comprise receptor-binding domains for cell surface proteins. Embodiment 115. The vaccine composition of embodiment 114, wherein the cell surface protein is mammalian. Embodiment 116. The vaccine composition of embodiment 113, wherein the receptor-binding domains of at least two of the six or more homologous distinct antigenic components share less than 99% sequence identity. Embodiment 117. The vaccine composition of embodiment 113, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 98% sequence identity. Embodiment 118. The vaccine composition of embodiment 113, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 95% sequence identity. Embodiment 119. The vaccine composition of embodiment 113, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 95% sequence identity. Embodiment 120. The vaccine composition of embodiment 113, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 90% sequence identity. Embodiment 121. The vaccine composition of embodiment 113, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 85% sequence identity. Embodiment 122. The vaccine composition of embodiment 113, wherein the receptor-binding domains of at least two of the six or more homologous distinct antigenic components share less than 80% sequence identity. Embodiment 123. The vaccine composition of embodiment 113, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 75% sequence identity. Embodiment 124. The vaccine composition of embodiment 113, wherein the receptor-binding domains of at least two of the six or more homologous distinct antigenic components share less than 70% sequence identity. Embodiment 125. A vaccine composition according to any one of embodiments 106 to 124, wherein administration of the vaccine composition reduces the predominance of immunogenic single mutant epitopes corresponding to the homologous distinct antigenic components. Embodiment 126. The vaccine composition of any one of embodiments 106 to 125, wherein any two of the six or more homologous different antigen components share less than about 90% sequence identity. Embodiment 127. The vaccine composition of embodiment 126, wherein any two of said six or more homologous different antigenic components share less than about 85% sequence identity. Embodiment 128. The vaccine composition of embodiment 127, wherein any two of said six or more homologous different antigenic components share less than about 80% sequence identity. Embodiment 129. The vaccine composition of embodiment 128, wherein any two of said six or more homologous different antigenic components share less than about 75% sequence identity. Embodiment 130. The vaccine composition of embodiment 129, wherein any two of said six or more homologous different antigenic components share less than about 70% sequence identity. Embodiment 131. The vaccine composition of any one of embodiments 106 to 130, wherein the vaccine composition comprises seven or more homologous different antigenic components. Embodiment 132. The vaccine composition of any one of embodiments 106 to 131, wherein the vaccine composition comprises 10 or more homologous different antigen components. Embodiment 133. The vaccine composition of any one of embodiments 106 to 132, wherein the vaccine composition comprises 15 or more homologous different antigen components. Embodiment 134. The vaccine composition of any one of embodiments 106 to 133, wherein the vaccine composition comprises 20 or more homologous different antigenic components. Embodiment 135. The vaccine composition of any one of embodiments 106 to 134, wherein the vaccine composition comprises 25 or more homologous different antigenic components. Embodiment 136. The vaccine composition of any one of embodiments 106 to 135, wherein the vaccine composition comprises 30 or more homologous different antigenic components. Embodiment 137. The vaccine composition of any one of embodiments 106 to 136, wherein any three of the six or more homologous different antigenic components share less than 95% sequence identity. Embodiment 138. The vaccine composition of any one of embodiments 106 to 137, wherein any four of the six or more homologous different antigenic components share less than 95% sequence identity. Embodiment 139. The vaccine composition of any one of embodiments 106 to 138, wherein any five of the six or more homologous different antigen components share less than 95% sequence identity. Embodiment 140. The vaccine composition of any one of embodiments 106 to 139, wherein any six of the six or more homologous different antigen components share less than 95% sequence identity. Embodiment 141. The vaccine composition of any one of embodiments 106 to 140, wherein any two of the six or more homologous different antigenic components share at least about 95% sequence identity. Embodiment 142. The vaccine composition of any one of embodiments 106 to 141, wherein any three of the six or more homologous different antigenic components share at least about 95% sequence identity. Embodiment 143. The vaccine composition of any one of embodiments 106 to 142, wherein any four of the six or more homologous different antigenic components share at least about 95% sequence identity. Embodiment 144. The vaccine composition of any one of embodiments 106 to 143, further comprising one or more heterologous antigen components. Embodiment 145. A vaccine composition according to any one of embodiments 106 to 144, further comprising an adjuvant. Embodiment 146. The vaccine composition of any one of embodiments 106 to 145, wherein the six or more homologous different antigenic components are present in the vaccine composition at the same concentration / amount. Embodiment 147. The vaccine composition of any one of embodiments 106 to 146, wherein the six or more homologous different antigenic components are present in the vaccine composition at different concentrations / amounts. Embodiment 148. The vaccine composition of any one of embodiments 106 to 147, wherein one dose of a homologous distinct antigenic component among the six or more homologous distinct antigenic components is calculated to be proportional to the phylogenetic distance between the homologous distinct antigenic component and another homologous distinct antigenic component, wherein the other homologous distinct antigenic component has the smallest distance to the homologous distinct antigenic component among the six or more homologous distinct antigenic components. Embodiment 149. The vaccine composition of any one of embodiments 106 to 148, wherein a dose of a homologous distinct antigenic component of the six or more homologous distinct antigenic components is calculated to be proportional to its average phylogenetic distance from other homologous distinct antigenic components of the six or more homologous distinct antigenic components. Embodiment 150. The vaccine composition of any one of embodiments 106 to 149, wherein the vaccine composition comprises a fragment of a SARS virus. Embodiment 151. A vaccine composition according to any one of embodiments 106 to 150, comprising a fragment of an influenza virus. Embodiment 152. The vaccine composition of any one of embodiments 106 to 151, wherein the vaccine composition comprises a fragment of the HIV virus. Embodiment 153. The vaccine composition of any one of embodiments 106 to 152, wherein the vaccine composition comprises a fragment of the SARS1 virus. Embodiment 154. A vaccine composition comprising six or more homologous different antigenic components, wherein two of the six or more homologous different antigenic components share less than 95% sequence identity; a. the homologous distinct antigenic component comprises a protein, and the concentration / amount of protein in a dose of the vaccine composition is from about 1 nanogram (ng) to about 3 micrograms (μg); b. the homologous distinct antigenic components comprise a plurality of RNAs, and the concentration / amount of RNA of the plurality of RNAs in a dose of the vaccine composition is from about 1 ng to about 5 μg per dose; or c. The homologous distinct antigenic components comprise multiple proteins displayed on heterologous virus-like particles (VLPs), and the concentration / amount of the multiple proteins displayed on the heterologous VLPs in a dose of the vaccine composition is from about 1 ng to about 5 μg per dose, and the vaccine composition is a veterinary vaccine composition. Embodiment 155. The vaccine composition of embodiment 154, wherein the vaccine composition is for administration to livestock animals. Embodiment 156 The vaccine composition of embodiment 155, wherein the vaccine composition is for administration to cows. Embodiment 157. The vaccine composition of embodiment 155, wherein the vaccine composition is for administration to sheep. Embodiment 158. The vaccine composition of embodiment 155, wherein the vaccine composition is for administration to pigs. Embodiment 159. The vaccine composition of embodiment 154, wherein the vaccine composition is for administration to birds. Embodiment 160. The vaccine composition of embodiment 154, wherein the vaccine composition is for administration to dogs. Embodiment 161 The vaccine composition of embodiment 154, wherein the vaccine composition is for administration to cats. Embodiment 162. The vaccine composition of any one of embodiments 154 to 161, wherein the concentration / amount of the homologous distinct antigenic component of the homologous distinct antigenic component in one dose of the vaccine composition is from about 1 nanogram (ng) to about 3 micrograms (μg). Embodiment 163. The vaccine composition of embodiment 162, wherein the concentration / amount of one of the homologous distinct antigenic components in one dose of the vaccine composition is from about 1 nanogram (ng) to about 1 microgram (μg). Embodiment 164. The vaccine composition of embodiment 163, wherein the concentration / amount of one of the homologous distinct antigenic components in one dose of the vaccine composition is from about 1 nanogram (ng) to about 0.1 microgram (μg). Embodiment 165. The vaccine composition of any one of embodiments 154 to 164, wherein the homologous distinct antigenic components comprise components of a virus. Embodiment 166. The vaccine composition of embodiment 165, wherein the viral component is a receptor-binding domain. Embodiment 167. The vaccine composition of embodiment 166, wherein at least two of the six or more homologous distinct antigenic components comprise receptor-binding domains for cell surface proteins. Embodiment 168. The vaccine composition of embodiment 167, wherein the cell surface protein is mammalian. Embodiment 169. The vaccine composition of embodiment 167, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 99% sequence identity. Embodiment 170. The vaccine composition of embodiment 169, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 98% sequence identity. Embodiment 171. The vaccine composition of embodiment 169, wherein the receptor-binding domains of at least two of the six or more homologous distinct antigenic components share less than 95% sequence identity. Embodiment 172. The vaccine composition of embodiment 169, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 95% sequence identity. Embodiment 173. The vaccine composition of embodiment 169, wherein the receptor-binding domains of at least two of the six or more homologous distinct antigenic components share less than 90% sequence identity. Embodiment 174. The vaccine composition of embodiment 169, wherein the receptor-binding domains of at least two of the six or more homologous distinct antigenic components share less than 85% sequence identity. Embodiment 175. The vaccine composition of embodiment 169, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 80% sequence identity. Embodiment 176. The vaccine composition of embodiment 169, wherein the receptor binding domains of at least two of the six or more homologous distinct antigenic components share less than 75% sequence identity. Embodiment 177. The vaccine composition of embodiment 169, wherein the receptor-binding domains of at least two of the six or more homologous distinct antigenic components share less than 70% sequence identity. Embodiment 178. A vaccine composition according to any one of embodiments 154 to 177, wherein administration of the vaccine composition reduces the predominance of immunogenic single mutant epitopes corresponding to the homologous distinct antigenic components. Embodiment 179. The vaccine composition of any one of embodiments 154 to 178, wherein any two of the six or more homologous different antigen components share less than about 90% sequence identity. Embodiment 180. The vaccine composition of embodiment 179, wherein any two of said six or more homologous different antigenic components share less than about 85% sequence identity. Embodiment 181. The vaccine composition of embodiment 180, wherein any two of said six or more homologous different antigenic components share less than about 80% sequence identity. Embodiment 182. The vaccine composition of embodiment 181, wherein any two of said six or more homologous different antigenic components share less than about 75% sequence identity. Embodiment 183. The vaccine composition of embodiment 182, wherein any two of said six or more homologous different antigenic components share less than about 70% sequence identity. Embodiment 184. The vaccine composition of any one of embodiments 154 to 183, wherein the vaccine composition comprises seven or more homologous different antigenic components. Embodiment 185. The vaccine composition of any one of embodiments 154 to 184, wherein the vaccine composition comprises 10 or more homologous different antigen components. Embodiment 186. The vaccine composition of any one of embodiments 154 to 185, wherein the vaccine composition comprises 15 or more homologous different antigen components. Embodiment 187. The vaccine composition of any one of embodiments 154 to 186, wherein the vaccine composition comprises 20 or more homologous different antigen components. Embodiment 188. The vaccine composition of any one of embodiments 154 to 187, wherein the vaccine composition comprises 25 or more homologous different antigenic components. Embodiment 189. The vaccine composition of any one of embodiments 154 to 188, wherein the vaccine composition comprises 30 or more homologous different antigenic components. Embodiment 190. The vaccine composition of any one of embodiments 154 to 189, wherein any three of the six or more homologous different antigenic components share less than 95% sequence identity. Embodiment 191. The vaccine composition of any one of embodiments 154 to 190, wherein any four of the six or more homologous different antigen components share less than 95% sequence identity. Embodiment 192. The vaccine composition of any one of embodiments 154 to 191, wherein any five of the six or more homologous different antigen components share less than 95% sequence identity. Embodiment 193. The vaccine composition of any one of embodiments 154 to 192, wherein any six of the six or more homologous different antigen components share less than 95% sequence identity. Embodiment 194. The vaccine composition of any one of embodiments 154 to 193, wherein any two of the six or more homologous different antigenic components share at least about 95% sequence identity. Embodiment 195. The vaccine composition of any one of embodiments 154 to 194, wherein any three of the six or more homologous different antigenic components share at least about 95% sequence identity. Embodiment 196. The vaccine composition of any one of embodiments 154 to 195, wherein any four of the six or more homologous different antigenic components share at least about 95% sequence identity. Embodiment 197. The vaccine composition of any one of embodiments 154 to 196, further comprising one or more heterologous antigen components. Embodiment 198. A vaccine composition according to any one of embodiments 154 to 197, further comprising an adjuvant. Embodiment 199. The vaccine composition of any one of embodiments 154 to 198, wherein the six or more homologous different antigenic components are present in the vaccine composition at the same concentration / amount. Embodiment 200. The vaccine composition of any one of embodiments 154 to 199, wherein the six or more homologous different antigenic components are present in the vaccine composition at different concentrations / amounts. Embodiment 201. The vaccine composition of any one of embodiments 154 to 200, wherein one dose of a homologous distinct antigenic component among the six or more homologous distinct antigenic components is calculated to be proportional to the phylogenetic distance between the homologous distinct antigenic component and another homologous distinct antigenic component, wherein the other homologous distinct antigenic component has the smallest distance to the homologous distinct antigenic component among the six or more homologous distinct antigenic components. Embodiment 202. The vaccine composition of any one of embodiments 154 to 201, wherein a dose of a homologous distinct antigenic component of the six or more homologous distinct antigenic components is calculated to be proportional to its average phylogenetic distance from other homologous distinct antigenic components of the six or more homologous distinct antigenic components. Embodiment 203. The vaccine composition of any one of embodiments 154 to 202, wherein the vaccine composition comprises a fragment of a SARS virus. Embodiment 204. A vaccine composition according to any one of embodiments 154 to 203, comprising a fragment of an influenza virus. Embodiment 205. The vaccine composition of any one of embodiments 154 to 204, wherein the vaccine composition comprises a fragment of the HIV virus. Embodiment 206. The vaccine composition of any one of embodiments 154 to 205, wherein the vaccine composition comprises a fragment of the SARS1 virus. Embodiment 207. A method of inducing an immune response against a pathogen in a subject, comprising administering a vaccine composition according to any one of embodiments 1 to 206. Embodiment 208. The method of embodiment 207, which is for prophylaxis against diseases caused by pathogens. [Example]

[0055] Example 1: Serum activity against mRNA antigens in mice

[0056] Mice were injected with either Centi-Flu (mRNA encoding a mixture of 16 hemagglutinin antigens, including eight H1 antigens corresponding to SEQ ID NOS: 4, 6, 8, 10, 12, 14, 16, and 18, and eight H3 antigens corresponding to SEQ ID NOS: 20, 22, 24, 26, 28, 30, 32, and 34) or Flu-Biv (mRNA encoding a mixture of two hemagglutinin antigens, including one H1 antigen (SEQ ID NOS: 10) and one H3 antigen (SEQ ID NOS: 22)). All mRNA vaccine compositions were complexed with lipid nanoparticles (LNPs). Specifically, the same LNP composition as for BNT162b2 was used. Serum was collected 16 days after immunization and measured by ELISA for antibodies binding to recombinant hemagglutinin proteins of H3N2 Hong Kong / 1 / 1968 (present in Centi-Flu), H3N2 California / 07 / 2004 (present in Centi-Flu and Flu-Biv), H3N2 Alaska / 01 / 2021 (present in Centi-Flu), H3N2 A / Victoria / 361 / 2011 (heterologous to both Centi-Flu and Flu-Biv), or H3N2 A / Maryland / 02 / 2021 (heterologous to both Centi-Flu and Flu-Biv). Figure 2A suggests that at the corresponding lower per-antigen dose, Flu-Biv did not induce a response, whereas Centi-Flu induced a robust response. For example, when mice were injected with 0.031 μg of Centi-Flu LNP per antigen (=0.5 μg total mRNA) or 0.031 μg of Flu-Biv per antigen (=0.063 μg total mRNA), Flu-Biv failed to induce a response, whereas Centi-Flu induced a robust response. Furthermore, 0.25 μg of Flu-Biv per antigen (=0.5 μg total mRNA) still induced weaker responses to all non-matching antigens than 0.031 μg of Centi-Flu per antigen (=0.5 μg total mRNA), so this difference cannot be explained by the total antigen dose. Figure 2B suggests that at higher doses per antigen, Flu-Biv induced weaker responses, whereas Centi-Flu induced stronger responses.For example, when mice were injected with 0.25 μg of Centi-Flu LNP per antigen (=4 μg total mRNA) or 0.25 μg of Flu-Biv per antigen (=0.5 μg total mRNA), Flu-Biv induced a weak response, while Centi-Flu induced a robust response. Furthermore, 2 μg of Flu-Biv per antigen (=4 μg total mRNA) still induced a weaker response to all non-matching antigens than 0.25 μg of Centi-Flu per antigen (=4 μg total mRNA), a difference that cannot be explained by the total antigen dose. Therefore, achieving the desired serological effect requires a unique combination of (1) six or more homologous, distinct antigen components and (2) low doses of mRNA encoding each component.

[0057] One exemplary set of eight homologous distinct antigenic components encoded by the mRNA of the Centi-Flu vaccine composition is A / Hong Kong / 1 / 1968 (SEQ ID NO:33), A / Nanchang / 933 / 1995 (SEQ ID NO:27), A / California / 07 / 2004 (SEQ ID NO:21), A / Memphis / 1 / 1980 (SEQ ID NO:29), A / Alaska / 01 / 2021 (SEQ ID NO:23), A / Indiana / 11 / 2018 (SEQ ID NO:19), A / Cambodia / e0826360 / 2020 (SEQ ID NO:25), and A / Bilthoven / 1971 / 1976 (SEQ ID NO:31). Figure 6 shows that any two of these eight H3N2 homologous distinct antigenic components have less than 96% and more than 80% pairwise sequence identity. Any subset of six of these eight constitutes a set of six homologous distinct antigenic components with pairwise sequence identity of less than 96% and greater than 80%.

[0058] mRNA production from DNA plasmids and subsequent encapsulation into lipid nanoparticles (LNPs) was performed using methods well known in the art and summarized herein. A DNA plasmid was constructed containing the same 5'UTR used in the clinically approved BNT162b2 vaccine (SEQ ID NO: 1), DNA encoding the antigen component, the same 3'UTR used in BNT162b2 (SEQ ID NO: 2), and a polyadenosine (120) tail under the SP6 promoter. The plasmid was linearized using the NotI restriction site and used as a template for in vitro transcription (IVT) using SP6 RNA polymerase and a mixture of nucleoside triphosphates. N1-methyl-pseudouridine-5'-triphosphate was used instead of uridine-5'-triphosphate for IVT. Capped mRNA was generated using the Vaccinia enzyme capping system (New England BioLabs). The purified mRNA was further encapsulated in the same LNP formulation as BNT162b2 (46.3% ionizable lipid ALC-0315, 9.4% phospholipid DSPC, 1.6% ALC-0159 PEG-lipid, and 42.7% cholesterol) to obtain LNP-encapsulated RNA.

[0059] Serum ELISA assays in all examples were performed as follows: The indicated proteins (5 μg / mL) were added to microtiter plates (CoStar) in coating buffer (0.1 M sodium bicarbonate, pH 8.6). After overnight incubation at 4°C and blocking with 3% bovine serum albumin (BSA) in PBS for 1 hour at 37°C, serum / plasma serially diluted in blocking buffer was added to individual wells and incubated for 1 hour at 37°C. The plates were then washed three times with 0.05% PBST. Horseradish peroxidase (HRP)-conjugated anti-IgG secondary antibody was added to the wells and incubated for 1 hour at 37°C. After washing three times with PBST, 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfone (Thermo Fisher Scientific Inc., Waltham, MA, USA) or 3,3′,5,5′-tetramethylbenzidine liquid substrate system (Thermo Fisher Scientific Inc.) was added to the wells. The absorbance was measured at 405 nm or 650 nm, respectively, using a microplate spectrophotometer (Multiskan GO, Thermo Fisher Scientific Inc.).

[0060] Example 2: Serum activity against mRNA antigens in ferrets

[0061] Ferrets were immunized once with mRNA encoding one of eight different antigens complexed with lipid nanoparticles (LNPs). Specifically, ferrets were immunized with 1 μg of LNP-encapsulated mRNA encoding hemagglutinin corresponding to H3N2 Alaska / 01 / 2021 (SEQ ID NO: 24), H3N2 California / 07 / 2004 (SEQ ID NO: 22), H3N2 Cambodia / 2020 (SEQ ID NO: 26), H3N2 Indiana / 11 / 2018 (SEQ ID NO: 20), H3N2 Bilthoven / 1761 / 1976 (SEQ ID NO: 32), H3N2 Nanchang / 933 / 1995 (SEQ ID NO: 28), or H3N2 Memphis / 1 / 1980 (SEQ ID NO: 30). Serum was obtained 28 days after immunization. Seroreactivity to the recombinant proteins of each antigen with which ferrets were immunized was measured as shown in Figure 3A. Seroreactivity corresponding to the antigen (i.e., sera from mice immunized with a given antigen tested against the same antigen) was very weak in all cases (EC50 was not achieved in any group, even at the highest tested serum dilution of 1:100).

[0062] Ferrets were immunized with LNP-encapsulated mRNA encoding hemagglutinins corresponding to eight H3N2 antigens (SEQ ID NOS: 24, 32, 22, 26, 20, 34, 28, and 30), each at approximately 0.5 μg per antigen (4 μg total H3N2 antigen). Serological reactivity to the recombinant proteins of each antigen with which ferrets were immunized was measured, as shown in Figure 3B. As shown in Figure 3B, ferrets immunized with Centi-Flu demonstrated strong responses to all immunizing antigens. This suggests that a single antigen at a low per-antigen dose (1 μg per antigen) is insufficient, but that Centi-Flu at an even lower per-antigen dose (0.5 μg per antigen) induces strong and broad reactivity. Thus, it is the unique combination of (1) six or more homologous, distinct antigenic components and (2) low doses of mRNA encoding each component that achieves the desired serological effect.

[0063] All LNP-encapsulated RNA constructs were produced as described in Example 1. Serum ELISA was performed as described in Example 1.

[0064] One exemplary set of eight homologous distinct antigenic components encoded by the mRNA of the Centi-Flu vaccine composition is A / Hong Kong / 1 / 1968 (SEQ ID NO:33), A / Nanchang / 933 / 1995 (SEQ ID NO:27), A / California / 07 / 2004 (SEQ ID NO:21), A / Memphis / 1 / 1980 (SEQ ID NO:29), A / Alaska / 01 / 2021 (SEQ ID NO:23), A / Indiana / 11 / 2018 (SEQ ID NO:19), A / Cambodia / e0826360 / 2020 (SEQ ID NO:25), and A / Bilthoven / 1971 / 1976 (SEQ ID NO:31). Figure 6 shows that any two of these eight H3N2 homologous distinct antigenic components have less than 96% and more than 80% pairwise sequence identity. Any subset of six of these eight constitutes a set of six homologous distinct antigenic components with pairwise sequence identity of less than 96% and greater than 80%.

[0065] Example 3: Serum activity against HIV protein antigens in mice

[0066] Centi-HIV is a mixture of gp120 / gp41 recombinant fusion proteins (linked by a flexible glycine-serine linker) from 10 different HIV antigens. Mice were immunized twice (days 0 and 14) with either Centi-HIV or a single gp120 / gp41 recombinant fusion protein ("single HIV antigen"). Serum was obtained on day 28. Serological reactivity to the same fusion proteins used in the "single HIV antigen" is shown in Figure 4. Figure 4 demonstrates that at a human equivalent (i.e., allometrically scaled) dose of 1 μg per antigen (=10 μg total human equivalent dose ("HED")), Centi-HIV elicits a robust response, while 1 μg of single HIV antigen fails to elicit reactivity even against the same antigens used for immunization. This cannot be explained by the total antigen dose, as 10 μg of single HIV antigen still initiates an inadequate response compared to Centi-HIV. Thus, it is the unique combination of (1) using six or more homologous distinct antigenic components and (2) administering each component at a low dose that achieves the desired serological effect.

[0067] Each antigen was recombinantly expressed in HEK293 cells by Sino Biological using methods well known in the art. The amino acid sequences of each immunogen are disclosed in SEQ ID NOs: 37, 41, 45, 49, 53, 57, 61, 65, 69, and 73. Each immunogen contains a given gp120 strain followed by a glycine-serine flexible linker (SEQ ID NO: 39), a given gp41 strain, a 3C protease cleavage site, a fold-on domain that promotes trimerization, and a C-terminal His tag. Serum ELISA was performed as described in Example 1. Figure 7 shows that any two of the 10 homologous distinct antigen components in the Centi-HIV composition share less than 80% pairwise sequence identity and more than 70% pairwise sequence identity.

[0068] Example 4: Seroreactivity to venom protein antigens in mice

[0069] Centi-Venom is a mixture of snake venom proteins from 15 different snake species (Agkistrodon contortrix contortrix, Agkistrodon piscivorus leucostoma, Agkistrodon piscivorus piscivorus, Agkistrodon bilineatus, Bitis arietans, Bothrops alternatus, Bothrops asper, Bothrops atrox, Bothrops jararaca, Crotalus atrox, Crotalus horridus, Daboia russelii, Daboia russelii limitis, Deinagkistrodon acutus, and Echis ocellatus). Mice were immunized six times at two-week intervals with either Centi-Venom or venom from a single snake species (single antigen) (Crotalus horridus). Serum was extracted from blood collected one week after the final immunization. Seroreactivity to snake venom from a single snake species (Crotalus horridus) is shown in Figure 5. Figure 5 demonstrates that Centi-Venom produces a strong response at a human equivalent (i.e., allometrically scaled) dose per antigen of 3 μg (=45 μg total human equivalent dose (“HED”)), while 3 μg of single antigen fails to elicit reactivity even against the same venom used for immunization. This cannot be explained by the total antigen dose, as 45 μg of single antigen still initiates an inadequate response compared to Centi-Venom. Therefore, it is the unique combination of (1) using six or more homologous, distinct antigenic components and (2) administering each component at a low dose that achieves the desired serological effect.

[0070] Serum ELISA was performed as described in Example 1. The amino acid sequences of the primary antigen (phospholipase A2; PLA2) in each snake venom are shown in SEQ ID NOs: 77-90. Figure 8 shows that any two of the homologous distinct antigenic components of the Centi-Venom composition have less than 85% pairwise sequence identity and more than 30% pairwise sequence identity.

[0071] In some embodiments, the invention includes a method of inducing an immune response against a pathogen in a subject, comprising administering a vaccine composition of the invention, hi some embodiments, the method is for prophylaxis against disease caused by the pathogen.

[0072] Whenever the terms "at least," "greater than," or "greater than or equal to" precede the first number in a series of two or more numbers, the terms "at least," "greater than," or "greater than or equal to" apply to each number in the series. For example, 1, 2, or 3 or more is equal to 1 or more, 2 or more, or 3 or more.

[0073] Whenever the term "less than or equal to," "less than," or "less than or equal to" precedes the first number in a series of two or more numbers, the term "less than or equal to," "less than," or "less than or equal to" applies to each number in the series. For example, 3, 2, or 1 or less is equivalent to 3 or less, 2 or less, or 1 or less.

[0074] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. It is not intended that the present invention be limited by the specific examples provided herein. While the present invention has been described with reference to the foregoing specification, the description and illustration of the embodiments herein are not meant to be construed in a limiting sense. Numerous variations, changes, and substitutions will occur to those skilled in the art without departing from the invention. Furthermore, it should be understood that all aspects of the present invention are not limited to the specific depictions, configurations, or relative proportions set forth herein, which depend upon a variety of conditions and variables. It should be understood that various alternatives to the embodiments of the present invention described herein may be used in practicing the present invention. It is therefore contemplated that the present invention also encompasses any such alternatives, modifications, variations, or equivalents. It is intended that the following claims define the scope of the invention, and that methods and structures within the scope of these claims and their equivalents be covered thereby. Specific Definitions

[0075] All terms are intended to be understood as understood by one of ordinary skill in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0076] The following definitions supplement the definitions in the art, and are directed to the current application, and should not be attributed to related or unrelated cases, such as jointly owned patents or applications. Although any method and material similar or equivalent to those described herein can be used to carry out the test of the present disclosure, preferred materials and methods are described herein.Therefore, the terms used herein are only intended to describe specific embodiments, and are not intended to be limiting.

[0077] The terms used herein are for the purpose of describing particular instances only and are not intended to be limiting. In this application, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the singular forms "a," "an," and "the" are intended to include the plural as well, unless the context clearly indicates otherwise. In this application, the use of "or" means "and / or" unless expressly stated otherwise. As used herein, the terms "and / or" and "any combination thereof," and their grammatical equivalents, may be used interchangeably. These terms may convey that any combination is specifically contemplated. For illustrative purposes only, the following phrases "A, B, and / or C" or "A, B, C, or any combination thereof" may mean "A alone, B alone, C alone, A and B, B and C, A and C, and A, B, and C." The term "or" may be used conjunctively or disjunctively unless the context specifically dictates a disjunctive use.

[0078] The terms "about" or "approximately" can mean within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean within one standard deviation or more than one standard deviation, in accordance with practice in the art. Alternatively, "about" can mean a range of up to 20%, up to 15%, up to 10%, up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, within five-fold, or within two-fold of a value. When particular values ​​are described in this application and claims, unless otherwise specified, the term "about" should be assumed to mean within an acceptable error range for the particular value.

[0079] As used in this specification and claims, the terms "comprising" (and any form of "comprise" or "comprises"), "having" (and any form of "have" or "has"), "including" (and any form of "includes" or "include"), or "containing" (and any form of "contains" or "contain") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. It is contemplated that any embodiment discussed herein can be implemented with respect to any method or composition of the disclosure, and vice versa. Furthermore, the compositions of the disclosure can be used to achieve the methods of the disclosure.

[0080] References herein to "some embodiments," "one embodiment," "an embodiment," or "other embodiments" mean that a particular feature, structure, or characteristic described in connection with an embodiment is included in at least some embodiments of the present disclosure, but not necessarily in all embodiments. To facilitate understanding of this disclosure, certain terms and phrases are defined below.

[0081] Ranges provided herein are understood to be shorthand for all values ​​within that range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or subrange from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50, as well as all intervening decimal values ​​between the aforementioned integers, such as, for example, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9. With respect to subranges, "nested subranges" extending from either end of the range are specifically contemplated. For example, nested subranges of the exemplary range of 1 to 50 could include 1 to 10, 1 to 20, 1 to 30, and 1 to 40 in one direction, or 50 to 40, 50 to 30, 50 to 20, and 50 to 10 in the other direction.

[0082] The term "pharmaceutically acceptable" refers to approved or approvable by a regulatory agency of a federal or state government, or listed in the United States Pharmacopeia (USP) or other generally recognized pharmacopeia for use in animals, including humans.

[0083] The term "subject" refers to an animal that has been the object of treatment, observation, or experiment. By way of example only, a subject includes, but is not limited to, a mammal, including a human or a non-human mammal, such as a non-human primate, cow, horse, dog, sheep, or cat.

[0084] The term "optional" or "optionally" means that the subsequently described event or circumstance can occur, but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not occur.

Claims

1. 1. A vaccine composition comprising: (a) a plurality of RNAs collectively encoding six or more homologous distinct antigenic components, wherein any two of the six or more homologous distinct antigenic components share less than 98% sequence identity, and wherein each RNA encoding a distinct antigenic component is present in the composition in an amount of 1 ng to 5 micrograms per dose; (b) a plurality of virus-like particles collectively displaying at least six or more homologous distinct antigenic components, wherein any two of the six or more homologous antigenic components share less than 98% sequence identity, and each distinct antigenic component is present in the composition in an amount of 1 ng to 5 micrograms; or (c) at least six or more homologous distinct antigenic components, wherein any two of the six or more homologous distinct antigenic components share less than 98% sequence identity, and each distinct antigenic component of the six or more homologous distinct antigenic components is present in the composition in an amount of 550 ng to 5 micrograms.

10. A vaccine composition comprising:

2. The vaccine composition of claim 1, wherein the vaccine composition is for the prevention of influenza.

3. A vaccine composition according to any one of claims 1 to 2, wherein each RNA encoding a different antigenic component is present in the composition in an amount of from 1 ng to 2.5 micrograms per dose.

4. A vaccine composition according to any one of claims 1 to 3, wherein each RNA encoding a different antigenic component is present in the composition in an amount of from 1 ng to 1.5 micrograms per dose.

5. A vaccine composition according to any one of claims 1 to 4, wherein each RNA encoding a different antigenic component is present in the composition in an amount of 1 ng to 1 microgram per dose.

6. 6. The vaccine composition of any one of claims 1 to 5, wherein each RNA encoding a different antigenic component is present in the composition in an amount of about 0.5 micrograms per dose.

7. A vaccine composition according to any one of claims 1 to 5, wherein each RNA encoding a different antigenic component is present in the composition in an amount of 1 microgram per dose.

8. A vaccine composition according to any one of claims 1 to 3, wherein each RNA encoding a different antigenic component is present in the composition in an amount of 2 micrograms per dose.

9. A vaccine composition according to any one of claims 1 to 2, wherein each RNA encoding a different antigenic component is present in the composition in an amount of from 1 ng to 4 micrograms per dose.

10. 10. The vaccine composition of any one of claims 1 to 9, wherein each RNA encoding a different antigenic component is present in the composition in an amount greater than 10 nanograms per dose.

11. 11. The vaccine composition of any one of claims 1 to 10, wherein each RNA encoding a different antigenic component is present in the composition in an amount greater than 100 nanograms per dose.

12. 12. The vaccine composition of any one of claims 1 to 11, wherein each RNA encoding a different antigenic component is present in the composition in an amount greater than 250 nanograms per dose.

13. 13. The vaccine composition of any one of claims 1 to 12, wherein each RNA encoding a different antigenic component is present in the composition in an amount greater than 500 nanograms per dose.

14. A vaccine composition according to any one of claims 1 to 2, wherein each different antigen component presented on a VLP is present in the composition in an amount of 1 ng to 2.5 micrograms per dose.

15. 15. The vaccine composition of any one of claims 1 to 2 or 14, wherein each different antigen component presented on a VLP is present in the composition in an amount of 1 ng to 1.5 micrograms per dose.

16. 16. A vaccine composition according to any one of claims 1 to 2 or 14 to 15, wherein each different antigen component presented on a VLP is present in the composition in an amount of 1 ng to 1 microgram per dose.

17. 17. The vaccine composition of any one of claims 1-2 or 14-16, wherein each different antigen component displayed on a VLP is present in the composition in an amount of about 0.5 micrograms per dose.

18. 3. The vaccine composition of any one of claims 1 to 2, wherein each different antigenic component of the six or more homologous different antigenic components is present in the composition in an amount of 550 ng to 2.5 micrograms per dose.

19. 20. The vaccine composition of any one of claims 1 to 2 or 18, wherein each different antigenic component of the six or more homologous different antigenic components is present in the composition in an amount of 550 ng to 1.5 micrograms per dose.

20. 20. The vaccine composition of any one of claims 1-2 or 18-19, wherein each different antigenic component of the six or more homologous different antigenic components is present in the composition in an amount of 550 ng to 1 microgram per dose.

21. 3. The vaccine composition of any one of claims 1 to 2, wherein each of the at least 6 or more homologous different antigenic components is present in the composition in an amount of 550ng to 3000ng.

22. 22. The vaccine composition of any one of claims 1 to 2 or 21, wherein each of the at least 6 or more homologous different antigenic components is present in the composition in an amount of 600ng to 3000ng.

23. 23. The vaccine composition of any one of claims 1-2 or 21-22, wherein each of the at least 6 or more homologous different antigenic components is present in the composition in an amount of 750ng to 3000ng.

24. 24. The vaccine composition of any one of claims 1 to 2 or 21 to 23, wherein each of the at least 6 or more homologous different antigenic components is present in the composition in an amount of 1000ng to 3000ng.

25. 24. The vaccine composition of any one of claims 1 to 2 or 21 to 23, wherein each of the at least 6 or more homologous different antigenic components is present in the composition in an amount of 750ng to 2000ng.

26. The vaccine composition of any one of claims 1 to 25, wherein the vaccine composition is for administration in a human subject.

27. 27. The vaccine composition of claim 26, wherein the vaccine composition is for administration to adults 18 years of age or older.

28. 27. The vaccine composition of claim 26, wherein the vaccine composition is for administration to adults 25 years of age or older.

29. 27. The vaccine composition of claim 26, wherein the vaccine composition is for administration to adults 50 years of age or older.

30. 27. The vaccine composition of claim 26, wherein the vaccine composition is for administration to adults 75 years of age or older.

31. 27. The vaccine composition of claim 26, wherein the vaccine composition is for administration to children aged from about 1 day to about 18 years.

32. 27. The vaccine composition of claim 26, wherein the vaccine composition is for administration to children aged from about 1 day to about 5 years.

33. 27. The vaccine composition of claim 26, wherein the vaccine composition is for administration to children between the ages of about 5 and about 18 years.

34. The vaccine composition according to any one of claims 1 to 17, wherein the vaccine composition is for administration to an animal.

35. 35. The vaccine composition of claim 34, wherein the animal is a livestock animal.

36. 37. The vaccine composition of claim 36, wherein the livestock animal is a cow, bull, alpaca, llama, sheep, pig, or bird.

37. 35. The vaccine composition of claim 34, wherein the animal is a domestic animal.

38. 38. The vaccine composition of claim 37, wherein the domestic animal is a primate.

39. 39. The vaccine composition of any one of claims 1 to 38, wherein the six or more homologous different antigenic components comprise antigens of viral, bacterial, fungal, prion or plant origin.

40. 40. The vaccine composition of any one of claims 1 to 39, wherein the homologous different antigenic components comprise components of a virus.

41. The vaccine composition of any one of claims 1 to 40, wherein the viral component is a receptor binding domain.

42. 42. The vaccine composition of any one of claims 1 to 41, wherein at least two of the six or more homologous distinct antigenic components comprise a receptor binding domain for a cell surface protein.

43. 43. The vaccine composition of claim 42, wherein the cell surface protein is mammalian.

44. 43. The vaccine composition of claim 42, wherein the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 99% sequence identity.

45. 43. The vaccine composition of claim 42, wherein the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 98% sequence identity.

46. 43. The vaccine composition of claim 42, wherein the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 95% sequence identity.

47. 43. The vaccine composition of claim 42, wherein the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 95% sequence identity.

48. 43. The vaccine composition of claim 42, wherein the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 90% sequence identity.

49. 43. The vaccine composition of claim 42, wherein the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 85% sequence identity.

50. 43. The vaccine composition of claim 42, wherein the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 80% sequence identity.

51. 43. The vaccine composition of claim 42, wherein the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 75% sequence identity.

52. 43. The vaccine composition of claim 42, wherein the receptor binding domains of any two of the six or more homologous distinct antigenic components share less than 70% sequence identity.

53. 53. The vaccine composition of any one of claims 1 to 52, wherein administration of said vaccine composition reduces the predominance of immunogenic single variant epitopes corresponding to said homologous distinct antigenic components.

54. 54. The vaccine composition of any one of claims 1 to 53, wherein any two of the six or more homologous different antigenic components share less than about 97.5% sequence identity.

55. 59. The vaccine composition of any one of claims 1 to 58, wherein any two of the six or more homologous different antigenic components share less than about 97% sequence identity.

56. 56. The vaccine composition of any one of claims 1 to 55, wherein any two of the six or more homologous different antigenic components share less than about 96% sequence identity.

57. 57. The vaccine composition of any one of claims 1 to 56, wherein any two of the six or more homologous different antigenic components share less than about 95% sequence identity.

58. 58. The vaccine composition of any one of claims 1 to 57, wherein any two of the six or more homologous different antigenic components share less than about 96% sequence identity.

59. 59. The vaccine composition of any one of claims 1 to 58, wherein any two of the six or more homologous different antigenic components share less than about 95% sequence identity.

60. 60. The vaccine composition of any one of claims 1 to 59, wherein any two of the six or more homologous different antigenic components share less than about 90% sequence identity.

61. 61. The vaccine composition of any one of claims 1 to 60, wherein any two of the six or more homologous different antigenic components share less than about 85% sequence identity.

62. 62. The vaccine composition of any one of claims 1 to 61, wherein any two of the six or more homologous different antigenic components share less than about 80% sequence identity.

63. 63. The vaccine composition of any one of claims 1 to 62, wherein any two of the six or more homologous different antigenic components share less than about 75% sequence identity.

64. 64. The vaccine composition of any one of claims 1 to 63, wherein any two of the six or more homologous different antigenic components share less than about 70% sequence identity.

65. 65. The vaccine composition of any one of claims 1 to 64, wherein the six or more homologous different antigenic components comprise seven or more homologous different antigenic components.

66. 66. The vaccine composition of any one of claims 1 to 65, wherein the six or more homologous different antigenic components comprises ten or more homologous different antigenic components.

67. 67. The vaccine composition of any one of claims 1 to 66, wherein the 6 or more homologous different antigenic components comprises 15 or more homologous different antigenic components.

68. 68. The vaccine composition of any one of claims 1 to 67, wherein the 6 or more homologous different antigenic components comprises 20 or more homologous different antigenic components.

69. 69. The vaccine composition of any one of claims 1 to 68, wherein the 6 or more homologous different antigenic components comprises 25 or more homologous different antigenic components.

70. 70. The vaccine composition of any one of claims 1 to 69, wherein the 6 or more homologous different antigenic components comprises 30 or more homologous different antigenic components.

71. 71. The vaccine composition of any one of claims 1 to 70, wherein any two of the six or more homologous different antigenic components share at least 30% sequence identity.

72. 72. The vaccine composition of any one of claims 1 to 71, wherein any two of the six or more homologous different antigenic components share at least 40% sequence identity.

73. 73. The vaccine composition of any one of claims 1 to 72, wherein any two of the six or more homologous different antigenic components share at least 50% sequence identity.

74. 74. The vaccine composition of any one of claims 1 to 73, wherein any two of the six or more homologous different antigenic components share at least 60% sequence identity.

75. 75. The vaccine composition of any one of claims 1 to 74, wherein any two of the six or more homologous different antigenic components share at least 70% sequence identity.

76. 76. The vaccine composition of any one of claims 1 to 75, wherein the vaccine composition further comprises an adjuvant.

77. 77. The vaccine composition of any one of claims 1 to 76, wherein the plurality of RNAs are present in the vaccine composition in the same amount.

78. 78. The vaccine composition of any one of claims 1 to 77, wherein the plurality of RNAs are present in the vaccine composition in different amounts.

79. 79. The vaccine composition of any one of claims 1 to 78, wherein the dose of one RNA of the plurality of RNAs is calculated to be proportional to the phylogenetic distance between a homologous distinct antigenic component of the six or more homologous distinct antigenic components and another homologous distinct antigenic component, wherein the other homologous distinct antigenic component has the smallest distance to the homologous distinct antigenic component among the six or more homologous distinct antigenic components.

80. 80. The vaccine composition of any one of claims 1 to 79, wherein the dose of one RNA of the plurality of RNAs is calculated to be proportional to the average phylogenetic distance of a homologous distinct antigenic component of the six or more homologous distinct antigenic components to other homologous distinct antigenic components of the six or more homologous distinct antigenic components.

81. 81. A method of inducing an immune response against an influenza pathogen in a subject, comprising administering a vaccine composition according to any one of claims 1 to 80.

82. 82. The method of claim 81 for prophylaxis against influenza.

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