DENV RNA VACCINES
Aspects of the disclosure relate to nucleic acid vaccines. The vaccines include one or more RNA polynucleotides having an open reading frame encoding one or more Chikungunya antigen(s), one or more Zika virus antigens, and one or more Dengue antigens. Methods for preparing and using such vaccines are also described.
1 .- 111 . (canceled)
112 . A Dengue virus (DENV) vaccine, comprising:
a ribonucleic acid (RNA) polynucleotide having an open reading frame encoding a DENV antigenic polypeptide.
113 . The DENV vaccine of claim 112 , wherein the DENV antigenic polypeptide is a DENV2 prME polypeptide.
114 . The DENV vaccine of claim 113 , wherein the DENV antigenic polypeptide is a DENV2 prME peptide epitope.
115 . The DENV vaccine of claim 113 , wherein the DENV antigenic polypeptide is a concatemeric DENV2 prME antigen.
116 . The DENV vaccine of claim 115 , wherein the concatemeric DENV2 prME antigen comprises 2-100 DENV2 prME peptide epitopes interspersed by cleavage sensitive sites.
117 - 118 . (canceled)
119 . The DENV vaccine of claim 114 , wherein the of the DENV2 prME peptide epitopes is a T cell epitope, a B cell epitope, or a combination of T cell epitopes and B cell epitopes.
120 .- 121 . (canceled)
122 . The DENV vaccine of claim 114 , wherein the DENV2 prME peptide epitope comprises a MHC class I epitope, a MHC class II epitope, or a combination of MHC class I epitopes and MHC class II epitopes.
123 .- 128 . (canceled)
129 . The DENV vaccine of claim 113 , wherein at least one of the DENV2 prME polypeptide comprises an amino acid sequence of SEQ ID NO: 15, 17, 19, 21, 23, 26, 29, 32, 169-297 or 298.
130 . The DENV vaccine of claim 112 , wherein the RNA polynucleotide further comprises a ubiquitination signal and/or an endosomal targeting sequence.
131 . (canceled)
132 . The DENV vaccine of claim 116 , wherein the cleavage sensitive sites are protease cleavage sites.
133 . The DENV vaccine of claim 132 , wherein the protease cleavage site is a cleavage site for a serine protease, a threonine protease, a cysteine protease, an aspartate protease, a glutamic acid protease, or a metalloprotease.
134 .- 139 . (canceled)
140 . The DENV vaccine of claim 113 , further comprising a RNA polynucleotide having an open reading frame encoding a DENV antigen selected from a DENV1, DENV3, DENV4, and/or DENV5 prME polypeptide.
141 .- 142 . (canceled)
143 . The DENV vaccine of claim 112 , wherein the open reading frame encodes at least two DENV antigenic polypeptides, or wherein the vaccine comprises at least two RNA polynucleotides each having an open reading frame encoding a DENV antigenic polypeptide.
144 .- 164 . (canceled)
165 . The DENV vaccine of claim 112 , wherein the antigenic polypeptide or immunogenic fragment thereof is obtained from a Brazilian DENV strain, ACS380, ACS46, ACS538, ACS542, ACS721, DGV106, DGV34, DGV37, DGV69, DGV91 or JHA1.
166 . The DENV vaccine of claim 112 , wherein the RNA polynucleotide comprises a chemical modification.
167 .- 168 . (canceled)
169 . The DENV vaccine of claim 166 , wherein the chemical modification is a N1-methylpseudouridine.
170 . The DENV vaccine of claim 166 , wherein at least 80% of the uracil in the open reading frame have a chemical modification.
171 . The DENV vaccine of claim 170 , wherein 100% of the uracil in the open reading frame have a chemical modification.
172 . The DENV vaccine of claim 112 , wherein the RNA polynucleotide further encodes a 5′ 7mG(5′)ppp(5′)NlmpNp terminal cap.
173 .- 177 . (canceled)
178 . The DENV vaccine of claim 112 , formulated in a lipid nanoparticle.
179 .- 180 . (canceled)
181 . The DENV vaccine of claim 178 , wherein the lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol and a non-cationic lipid.
182 . The DENV vaccine of claim 181 , wherein the lipid nanoparticle carrier comprises a molar ratio of about 20-60% cationic lipid, 0.5-15% PEG-modified lipid, 25-55% sterol, and 25% non-cationic lipid.
183 . The DENV vaccine of claim 181 , wherein the cationic lipid is an ionizable cationic lipid and the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol.
184 . The DENV vaccine of claim 181 , wherein the cationic lipid is selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319).
185 .- 190 . (canceled)
191 . A method of inducing an immune response in a subject, the method comprising administering to the subject the DENV vaccine of claim 112 in an amount effective to produce an antigen-specific immune response in the subject.
192 .- 237 . (canceled)
238 . A Dengue virus (DENV) vaccine, comprising a ribonucleic acid (RNA) polynucleotide having an open reading frame encoding a DENV antigenic polypeptide, a 5′ terminal cap, and a chemical modification, wherein the vaccine is formulated within a lipid nanoparticle.
239 . The DENV vaccine of claim 238 , wherein at least 80% of the uracil in the open reading frame have a chemical modification.
240 .- 244 . (canceled)
245 . A Dengue virus (DENV) vaccine, comprising a ribonucleic acid (RNA) polynucleotide having an open reading frame encoding a DENV antigenic peptide linked to a signal peptide.
246 . The DENV vaccine of claim 245 , wherein the DENV antigenic polypeptide is a DENV prM protein, a DENV envelope (E) protein, or a DENV prME fusion protein.
247 .- 375 . (canceled)
376 . The vaccine of claim 112 , further comprising a RNA polynucleotide having an open reading frame encoding a CHIKV antigenic polypeptide and/or a ZIKV antigenic polypeptide.
377 .- 477 . (canceled)