CN-121991183-A - Bovine parainfluenza virus 3A and 3C multi-epitope antigen peptide, and complexes and application thereof

Abstract

The invention discloses bovine parainfluenza virus 3A and 3C multi-epitope antigen peptide, a complex thereof and application thereof. The amino acid sequences of the multi-epitope antigen peptides BPMEV-3A and BPMEV-3C are respectively shown as SEQ ID NO. 1 and SEQ ID NO. 3, and are formed by connecting CTL epitopes, HTL epitopes and B cell epitopes screened from HN and F proteins of BPIV-3A, BPIV-3C strains. Animal immunity experiments show that the antigen peptide and the compound thereof can effectively stimulate organisms to generate specific IgG antibodies and neutralizing antibodies, induce Th1 type cell immune response, effectively remove viruses, alleviate lung tissue lesions and show good immunogenicity and protection effects. The invention provides efficient and safe vaccine candidates for prevention and control of BPIV-3, and has good application prospect.

Inventors

• GUO AIZHEN
• GUO LEI
• JI WANFENG
• CHEN JIANGUO
• ZHANG LEI
• ZHAO MENGYING
• CHEN YINGYU
• CHEN XI
• XU XIAOWEN
• WU WENYING
• YANG CHAORAN
• CHEN SIYU
• QI ZIJUN

Assignees

• 华中农业大学

Dates

Publication Date

20260508

Application Date

20260410

Claims (9)

1. 1. The amino acid sequence of the bovine parainfluenza virus 3A type multi-epitope antigen peptide is shown as SEQ ID NO. 1.
2. 2. The nucleotide sequence of the gene for encoding bovine parainfluenza virus 3A type multi-epitope antigen peptide is shown as SEQ ID NO. 2.
3. 3. The amino acid sequence of the bovine parainfluenza virus 3C type multi-epitope antigen peptide is shown as SEQ ID NO. 3.
4. 4. A gene encoding the bovine parainfluenza virus 3C multi-epitope antigen peptide of claim 3, the nucleotide sequence of which is shown in SEQ ID No. 4.
5. 5. An antigenic peptide complex comprising the bovine parainfluenza virus 3A-type multi-epitope antigenic peptide of claim 1 and the bovine parainfluenza virus 3C-type multi-epitope antigenic peptide of claim 3.
6. 6. Use of the antigenic peptide of claim 1 or 3 or the antigenic peptide complex of claim 5 in the manufacture of a kit for detecting bovine parainfluenza virus type 3.
7. 7. A kit for detecting bovine parainfluenza virus type 3 comprising the antigenic peptide of claim 1 or 3 or the antigenic peptide complex of claim 5.
8. 8. Use of the antigenic peptide of claim 1 or 3 or the antigenic peptide complex of claim 5 for the preparation of a bovine parainfluenza virus type 3 vaccine.
9. 9. A bovine parainfluenza virus type 3 vaccine comprising the antigenic peptide of claim 1 or 3 or the antigenic peptide complex of claim 5.

Description

Bovine parainfluenza virus 3A and 3C multi-epitope antigen peptide, and complexes and application thereof Technical Field The invention belongs to the field of biology, relates to bovine parainfluenza virus 3A and 3C multi-epitope antigen peptides and application thereof, and also relates to an antigen peptide complex and application thereof, in particular to application in preparation of bivalent vaccine. Background Bovine parainfluenza virus type 3 (Bovine parainfluenza-3 virus, bpiv-3), belonging to the genus respiratory virus of the family paramyxoviridae, is the primary pathogen responsible for bovine parainfluenza (Bovine parainfluenza, BPI). The disease is an acute contagious disease, can independently cause bovine respiratory disease, can also cause bovine respiratory syndrome (Bovine respiratory disease complex, BRDC) by mixed infection of mycoplasma bovis, pasteurella multocida A-type and other pathogens, and causes serious economic loss to the global cattle raising industry. With the large-scale development of cattle industry, effective prevention and control of BPI is particularly important, and vaccination is the most effective prevention and control strategy at present. BPIV-3 is an enveloped, non-segmented, single-stranded negative-strand RNA virus. BPIV-3 is classified into A, B, C genotypes according to the nucleotide homology of HN gene, and the major popular genotypes in China are BPIV-3A type and BPIV-3C type. HN protein and F protein on the surface of the virus are main antigen proteins of the virus, can induce the organism to generate neutralizing antibodies, and are core targets of vaccine design. The BPIV-3 vaccine commercialized at present mainly comprises an inactivated vaccine and a attenuated vaccine. However, these traditional vaccines have the respective limitations of good safety of inactivated vaccines but poor capability of inducing cellular immunity, and strong immunogenicity of attenuated vaccines but potential risk of virulence return. Furthermore, because BPIV-3 is subject to genetic mutation and antigenic drift under evolutionary pressure, it is often difficult for a single subtype of vaccine to provide effective cross-protection for different subtypes of epidemic strains. Epitope vaccines have been developed rapidly in recent years as a novel vaccine. The technology can induce high-specificity immune response aiming at humoral immunity and cellular immunity simultaneously by screening multiple types of immunogenic epitopes in antigen proteins and constructing the multiple epitope peptide antigens in series. Compared with the traditional vaccine, the multi-epitope vaccine has higher flexibility in design, can optimize the most protective epitope according to the evolution characteristics of the virus, and is particularly suitable for the virus with highly variable genome. Although studies have been made to predict epitopes of BPIV-3 using bioinformatics, most of these studies remain in the theoretical prediction stage, and no specific, experimentally verified multi-epitope antigenic peptide sequences capable of simultaneously targeting both BPIV-3A and BPIV-3C epidemic strains have been disclosed. Therefore, there is a need in the art to develop a BPIV-3A and 3C bivalent multi-epitope antigen peptide having a definite amino acid sequence, which can be efficiently expressed by a prokaryotic system, and which has good immunogenicity as verified by animal experiments, and based on which a high-efficiency, safe vaccine capable of covering various subtypes is prepared. Based on the two novel multi-epitope antigen peptides BPMEV-3A and BPMEV-3C are provided for the first time through bioinformatics screening, molecular docking design, prokaryotic expression purification and animal experiment verification, and the bivalent vaccine formed by combining the two novel multi-epitope antigen peptides BPMEV-3A and BPMEV-3C is proved to have good immune protection effect, and important research value and application prospect are achieved. Disclosure of Invention The first object of the invention is to provide bovine parainfluenza virus type 3A and type 3C multi-epitope antigen peptides. When the multi-epitope antigen peptide is constructed, CTL epitopes, HTL epitopes and B cell epitopes are fused, and the synergistic effect of the three epitopes can simulate the complete pathogen infection process, comprehensively activate the adaptive immune system of organisms and realize double protection of humoral immunity and cellular immunity. By optimizing the connection mode and sequence of the linker, each surface can be ensured to be correctly processed, presented and identified, and finally, balanced and durable humoral immunity and cellular immunity response are induced, so that comprehensive and effective protection is provided for a host. The amino acid sequence of the bovine parainfluenza virus 3A type multi-epitope antigen peptide is shown as SEQ ID NO. 1, and the nucleotide sequence