Antigenic chimeric tick-borne encephalitis virus/dengue virus type 4 recombinant viruses

摘要

Disclosed herein are chimeric TBEV/DEN4 flaviviruses including a first nucleic acid molecule including a 5′ non-coding region (NCR) from a DEN4 virus, a nucleic acid encoding a C protein and non-structural proteins from a DEN4 virus, and a 3′ NCR from a DEN4 virus, wherein nonstructural protein NS4B includes a phenylalanine at amino acid position 112, nonstructural protein NS5 includes an alanine at amino acid position 654 and an alanine at amino acid position 655, and the 3′ NCR includes a deletion of nucleotides 10478-10507. The chimeric construct also includes a second nucleic acid molecule, which is operably linked to the first nucleic acid molecule, encoding a prM protein and an E protein from a TBEV, wherein the E protein includes an amino acid substitution that differs from the wild type TBEV at amino acid position 315 and a tryptophan at amino acid position 240. Also disclosed are methods of eliciting an immune response using the disclosed TBEV/DEN4 chimeric flaviviruses and immunogenic compositions including the disclosed chimeric flaviviruses and a pharmaceutically acceptable carrier.

主权项

1. A recombinant chimeric virus comprising:
a first nucleic acid molecule comprising:
a 5′ non-coding region from a dengue type 4 virus;a nucleic acid encoding non-structural proteins from a dengue type 4 virus, wherein nonstructural protein NS4B comprises a phenylalanine at amino acid position 112 corresponding to amino acid position 2359 of SEQ ID NO: 2, and nonstructural protein NS5 comprises an alanine at amino acid position 654 corresponding to amino acid position 3146 of SEQ ID NO: 2 and an alanine at amino acid position 655 corresponding to amino acid position 3147 of SEQ ID NO: 2; and
a 3′ non-coding region from a dengue type 4 virus, wherein the 3′ non-coding region comprises a deletion of nucleotides corresponding to nucleotides 10478-10507 of SEQ ID NO: 27; and a second nucleic acid molecule operably linked to the first nucleic acid molecule, the second nucleic acid molecule encoding:
a prM protein from a tick borne encephalitis virus; andan E protein from a tick borne encephalitis virus, wherein the E protein comprises an amino acid substitution from wild-type at amino acid position 315 corresponding to amino acid position 598 of SEQ ID NO: 2 and a tryptophan at amino acid position 240 corresponding to amino acid position 523 of SEQ ID NO: 2, wherein the chimeric virus does not encode a C protein.