BROAD-SPECTRUM MULTI-ANTIGEN PAN-CORONAVIRUS VACCINE
Waning immunity induced by first-generation Spike-alone-based COVID-19 has failed to prevent immune escape by many variants of concern (VOCs) that emerged from 2020 to 2024, resulting in a prolonged COVID-19 pandemic. Thus, a next-generation Coronavirus (CoV) vaccine incorporating highly conserved non-Spike SARS-CoV-2 antigens is described herein. Conserved non-Spike T cell antigens in combination with a Spike antigen encapsulated in lipid nanoparticles: (i) Induced high frequencies of lung-resident antigen-specific CXCR5+CD4+ T follicular helper cells, GzmB+CD4+ and GzmB+CD8+ cytotoxic T cells, and CD69+IFN-γ+TNFα+CD4+ and CD69+IFN-γ+TNFα+CD8+ effector T cells; and (ii) Reduced viral load and COVID-19-like symptoms caused by various VOCs. The combined antigen/LNP-based pan-CoV vaccine could be rapidly adapted for clinical use to confer broader cross-protective immunity against emerging highly mutated and pathogenic VOCs.
1 .- 24 . (canceled)
25 . A composition, comprising a lipid nanoparticle, wherein the lipid nanoparticle comprises a first messenger ribonucleic acid (mRNA) encapsulated therein, a second mRNA encapsulated therein, and a third mRNA encapsulated therein, wherein:
the first mRNA comprises an open reading frame encoding a single Coronavirus NSP14 protein;
the second mRNA comprises an open reading frame encoding a single Coronavirus Nucleoprotein; and
the third mRNA comprises an open reading frame encoding a single Coronavirus NSP2 protein.
26 . The composition of claim 25 , wherein each mRNA further comprises a 5′ untranslated region (UTR) and a 3′ UTR.
27 . The composition of claim 25 , wherein each mRNA further comprises a poly(A) tail and a 5′ cap.
28 . The composition of claim 25 , wherein the lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid.
29 . The composition of claim 28 , wherein the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol.
30 . The composition of claim 29 , wherein the cationic lipid is selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane, dilinoleyl-methyl-4-dimethylaminobutyrate, or di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate.
31 . A composition, comprising a lipid nanoparticle, wherein the lipid nanoparticle comprises a first messenger ribonucleic acid (mRNA) encapsulated therein, a second mRNA encapsulated therein, and a third mRNA encapsulated therein, wherein:
the first mRNA comprises an open reading frame encoding a single Coronavirus NSP14 protein encoded by SEQ ID NO:4 or SEQ ID NO:5;
the second mRNA comprises an open reading frame encoding a single Coronavirus Nucleoprotein encoded by SEQ ID NO:7 or SEQ ID NO:8; and
the third mRNA comprises an open reading frame encoding a single Coronavirus NSP2 protein encoded by SEQ ID NO:1 or SEQ ID NO:2.
32 . The composition of claim 31 , wherein each mRNA further comprises a 5′ untranslated region (UTR) and a 3′ UTR.
33 . The composition of claim 31 , wherein each mRNA further comprises a poly(A) tail and a 5′ cap.
34 . The composition of claim 31 , wherein the lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid.
35 . The composition of claim 34 , wherein the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol.
36 . The composition of claim 35 , wherein the cationic lipid is selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane, dilinoleyl-methyl-4-dimethylaminobutyrate, or di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate.