Vaccine compositions against porcine reproductive and respiratory syndrome and porcine circovirus associated diseases
Inventors
CHIEN, Yu-Hsin • Tsai, Meng-Ju • LAI, Pao-Yen • Chou, Wei-I • Chang, Hsiu-Kang
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-10010602-B2
Publication Date
2018-07-03
Expiration Date
Abstract
A fusion protein comprising an antigen-presenting cell (APC)-binding domain or a CD91 receptor-binding domain, a translocation peptide, a fusion antigen, an endoplasmic reticulum retention sequence, and optionally a nuclear export signal is disclosed. The fusion antigen comprises a porcine reproductive and respiratory syndrome virus (PRRSV) ORF7 antigen, a PRRSV ORF1b antigen, a PRRSV ORF6 antigen, and a PRRSV ORF5 antigen. The fusion protein is useful for inducing antigen-specific cell-mediated and humoral responses.
Core Innovation
A fusion protein is described for use as a PRRSV vaccine fusion protein. The fusion protein comprises an antigen-presenting cell (APC)-binding domain at the N-terminus, wherein the APC-binding domain is a Pseudomonas exotoxin A (PE) binding domain. A translocation peptide of 34-112 amino acid residues, comprising an amino acid sequence of SEQ ID NO: 4, 2, 3, or 6, is located at the C-terminus of the APC-binding domain.
The fusion protein further comprises a fusion antigen that includes a PRRSV ORF7 antigen, a PRRSV ORF1b antigen, a PRRSV ORF6 antigen, and a PRRSV ORF5 antigen. The fusion antigen does not comprise full-length ORF7, ORF6, ORF5, and ORF1b protein sequences. A nuclear export signal comprising the amino acid sequence of SEQ ID NO: 13 is located at the C-terminus of the fusion antigen or between the translocation peptide and the fusion antigen.
An endoplasmic reticulum retention sequence is also included, positioned at the C-terminus of the fusion antigen when the nuclear export signal is located between the translocation peptide and the fusion antigen, or positioned at the C-terminus of the nuclear export signal when the nuclear export signal is located at the C-terminus of the fusion antigen. The document further describes an optional inclusion of a PCV2 fusion protein component with a C-terminal-truncated PCV2 ORF2 antigen as part of vaccine formulations.
Claims Coverage
The claims cover one fusion protein architecture that combines a PE-based APC-binding domain, a 34-112 amino acid translocation peptide, a PRRSV multi-antigen fusion antigen based on ORF7/ORF1b/ORF6/ORF5 without full-length sequences, and trafficking signals including a nuclear export signal and an endoplasmic reticulum retention sequence. Additional claim elements specify subranges and motif constraints and include an administration-based immunogenicity use, with optional extension to PCV2 fusion protein components in compositions.
PE binding domain–based APC-targeting fusion protein architecture
An antigen-presenting cell (APC)-binding domain located at the N-terminus of the fusion protein, wherein the APC-binding domain is a Pseudomonas exotoxin A (PE) binding domain.
C-terminal translocation peptide (34-112 aa; SEQ ID options)
A translocation peptide of 34-112 amino acid residues in length, comprising the amino acid sequence of SEQ ID NO: 4, 2, 3, or 6, located at the C-terminus of the APC-binding domain.
PRRSV multi-antigen fusion antigen without full-length ORF proteins
A fusion antigen comprising a PRRSV ORF7 antigen, a PRRSV ORF1b antigen, a PRRSV ORF6 antigen, and a PRRSV ORF5 antigen, wherein the fusion antigen does not comprise full-length ORF7, ORF6, ORF5, and ORF1b protein sequences.
Nuclear export signal placement with ER retention sequence positioning
A nuclear export signal comprising the amino acid sequence of SEQ ID NO: 13, located at the C-terminus of the fusion antigen or between the translocation peptide and the fusion antigen, and an endoplasmic reticulum retention sequence positioned at the C-terminus of the fusion antigen when the nuclear export signal is located between the translocation peptide and the fusion antigen, or positioned at the C-terminus of the nuclear export signal when the nuclear export signal is located at the C-terminus of the fusion antigen.
Therapeutically effective administration to induce antigen-specific immunity
A method of inducing antigen-specific cell-mediated and humoral immune responses by administering to a subject a therapeutically effective amount of the fusion protein of claim 1.
Optional PCV2 fusion protein component with constrained PCV2 ORF2 antigen
A composition including the fusion protein of claim 1 together with a porcine circovirus type 2 (PCV2) fusion protein engineered with a PCV2 ORF2 antigen containing only the C-terminal portion of PCV2 ORF2 protein, together with trafficking-signal features including a nuclear export signal comprising the amino acid sequence of SEQ ID NO: 13 and an endoplasmic reticulum retention sequence positioned based on the nuclear export signal placement.
Overall, the claims center on a PRRSV multi-antigen fusion protein that uses a PE binding domain at the N-terminus, a 34-112 aa PE-related translocation peptide at the C-terminus of that domain, and truncated PRRSV ORF7/ORF1b/ORF6/ORF5 fusion antigen components, combined with a nuclear export signal (SEQ ID NO: 13) and an endoplasmic reticulum retention sequence whose position depends on the nuclear export signal location.
Stated Advantages
Stronger antigen-specific IFN-γ (CMI) responses and IgG (humoral) responses compared with PRRSFREE 4-in-1 single-fusion versus multi-antigen formulations, as described in the document.
Combination vaccine comparisons show reductions in swine viremia after PRRSV challenge, as described in the document.
Documented Applications
Vaccine formulations using the described PRRSV fusion proteins in mice and swine, with reported immunogenicity (antigen-specific IFN-γ and IgG) and PRRSV challenge outcome measurements including swine viremia reduction.
Use in combination vaccine comparisons including PRRSV/PCV2 formulations, as described in the document.
Interested in licensing this patent?