Nucleic acids encoding chimeric dengue/Zika viruses optimized for growth and stability in vero cells
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services
InventorClaire Y. H. Kinney
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Abstract
Chimeric flaviviruses that include non-coding regions, non-structural proteins, a capsid (C) protein and a portion of a premembrane (prM) signal sequence from an attenuated or wild-type dengue serotype 2 virus (DENV-2), and a portion of a prM signal sequence, a prM protein and at least a portion of an envelope (E) protein from a Zika virus (ZIKV) are described. Also described are immunogenic compositions and methods for eliciting an immune response in a subject, such as an immune response directed against ZIKV.
Claims27
A nucleic acid chimera encoding a chimeric dengue virus serotype 2 (DENV-2)/Zika virus (ZIKV), wherein the ZIKV premembrane (prM) and envelope (E) genes are inserted into the genomic background of a DENV-2 strain, wherein the nucleic acid chimera comprises the following genomic arrangement:
- DENV-2 5′ non-coding region, DENV-2 capsid (C) gene, ZIKV prM/E genes, DENV-2 non-structural (NS) 1-NS5 genes, and DENV-2 3′ non-coding region, wherein:
- the nucleic acid chimera encodes a modified prM signal sequence comprising the first three amino acids of the DENV-2 prM signal sequence fused to the last 15 amino acids of the ZIKV prM signal sequence;
- or the first five amino acids of the DENV-2 prM signal sequence fused to the last 13 amino acids of the ZIKV prM signal sequence;
- the nucleic acid chimera encodes a chimeric ZIKV/DENV-2 E protein, wherein the last 14 amino acids of the E protein are from DENV-2;
- and
- the nucleic acid chimera comprises:
- (i) four Vero cell adaptation mutations, wherein the mutations result in:
- a glutamine to arginine substitution at residue 465 of the E protein;
- an isoleucine to phenylalanine substitution at residue 493 of the E protein;
- a lysine to asparagine substitution at residue 99 of the NS2A protein;
- and
- an aspartic acid to asparagine substitution at residue 23 of the NS4A protein;
- (ii) four Vero cell adaptation mutations, wherein the mutations result in:
- a glutamine to arginine substitution at residue 465 of the E protein;
- an isoleucine to threonine substitution at residue 484 of the E protein;
- an isoleucine to phenylalanine substitution at residue 493 of the E protein;
- and
- a lysine to asparagine substitution at residue 99 of the NS2A protein;
- or
- (iii) five Vero cell adaptation mutations, wherein the mutations result in:
- a glutamine to arginine substitution at residue 465 of the E protein;
- an isoleucine to threonine substitution at residue 484 of the E protein;
- an isoleucine to phenylalanine substitution at residue 493 of the E protein;
- a lysine to asparagine substitution at residue 99 of the NS2A protein;
- and
- an aspartic acid to asparagine substitution at residue 23 of the NS4A protein.
A nucleic acid chimera encoding a chimeric dengue virus serotype 2 (DENV-2)/Zika virus (ZIKV), wherein the ZIKV premembrane (prM) and envelope (E) genes are inserted into the genomic background of a DENV-2 strain, wherein the nucleic acid chimera comprises the following genomic arrangement:
- DENV-2 5′ non-coding region, DENV-2 capsid (C) gene, ZIKV prM/E genes, DENV-2 non-structural (NS) 1-NS5 genes, and DENV-2 3′ non-coding region, wherein:
- the nucleic acid chimera encodes a modified prM signal sequence comprising the first three amino acids of the DENV-2 prM signal sequence fused to the last 15 amino acids of the ZIKV prM signal sequence;
- or the first five amino acids of the DENV-2 prM signal sequence fused to the last 13 amino acids of the ZIKV prM signal sequence;
- the nucleic acid chimera encodes a chimeric ZIKV/DENV-2 E protein, wherein the last 13 amino acids of the E protein are from DENV-2;
- and
- the nucleic acid chimera comprises:
- (i) four Vero cell adaptation mutations, wherein the mutations result in:
- a glutamine to arginine substitution at residue 465 of the E protein;
- an isoleucine to phenylalanine substitution at residue 493 of the E protein;
- a lysine to asparagine substitution at residue 99 of the NS2A protein;
- and
- an aspartic acid to asparagine substitution at residue 23 of the NS4A protein;
- (ii) four Vero cell adaptation mutations, wherein the mutations result in:
- a glutamine to arginine substitution at residue 465 of the E protein;
- an isoleucine to threonine substitution at residue 484 of the E protein;
- an isoleucine to phenylalanine substitution at residue 493 of the E protein;
- and
- a lysine to asparagine substitution at residue 99 of the NS2A protein;
- or
- (iii) five Vero cell adaptation mutations, wherein the mutations result in:
- a glutamine to arginine substitution at residue 465 of the E protein;
- an isoleucine to threonine substitution at residue 484 of the E protein;
- an isoleucine to phenylalanine substitution at residue 493 of the E protein;
- a lysine to asparagine substitution at residue 99 of the NS2A protein;
- and
- an aspartic acid to asparagine substitution at residue 23 of the NS4A protein.
Dependencies are inferred from the claim text.