38 results about "B-Cell Epitopes" patented technology

The invention discloses a conservative B cell epitope peptide of goose parvovirus VP3 protein, a nucleic acid molecule, a recombinant vector and application thereof. Conservative B cell epitope EW12 is inserted into F18 Escherichia coli Fod fimbriae, inert carrier bacteria S9H are introduced, recombinant bacteria capable of functionally exhibiting and expressing the EW12 conservative B cell epitope on the surface of the bacteria are obtained, the recombinant bacteria can specifically generate specific binding reaction with gosling plague egg yolk antibody and gosling plague vaccine immune serum, and the gosling plague egg yolk antibody and the gosling plague vaccine immune serum can be specifically combined with the gosling plague egg yolk antibody and the gosling plague vaccine immune serum. The method has the advantages that macroscopic agglutination particles are generated, cross agglutination reaction with other pathogen positive serum and healthy goose serum is avoided, the method is good in sensitivity, and the specific antibody can be detected in the 5th day after immunization at the earliest. A plate agglutination test directly mediated by the goose parvovirus VP3 protein conservative B cell epitope peptide EW12 has the advantages of specificity, sensitivity, convenience in operation and suitability for on-site large-scale rapid detection, and is expected to provide a new technical means for diagnosis, prevention and control and purification of GPV infection.

The invention provides a vaccine composition comprising an influenza virus peptide comprising a CD8+ T cell epitope and an influenza virus peptide comprising a B cell epitope, wherein each peptide is attached to a nanoparticle.

The invention belongs to the technical field of genetic engineering, and particularly relates to a bovine group A rotavirus multi-epitope fusion protein and application thereof. The invention provides a bovine group A rotavirus multi-epitope peptide based on a ferritin nano-carrier, a fusion protein and application of the bovine group A rotavirus multi-epitope peptide and the fusion protein. The multi-epitope peptide disclosed by the invention is a multi-epitope fusion antigen which is formed by splicing cytotoxic T lymphocyte epitopes, helper T cell epitopes and B cell epitopes with high conservative property and strong immunogenicity and is formed by screening and obtaining the cytotoxic T lymphocyte epitopes, helper T cell epitopes and B cell epitopes with high conservative property and strong immunogenicity on the basis of VP4 and VP7 protein sequences of bovine group A rotaviruses by utilizing an immunoinformatics technology. Immunological evaluation shows that the monoclonal antibody has good antigen specificity and neutralizing activity. Animal experiment results show that the epitope peptide and the fusion protein can induce an organism to generate high-level neutralizing antibodies aiming at BRVA G6, G8, G10 and other multi-genotype strains, and the broad spectrum and durability of immune protection are remarkably improved.

The application discloses a bovine parainfluenza virus 3A and 3C type polyepitope antigen peptide, a complex thereof and application. The polyepitope antigen peptides BPMEV-3A and BPMEV-3C have amino acid sequences as shown in SEQ ID NO:1 and SEQ ID NO:3 respectively, and are connected by screening CTL epitopes, HTL epitopes and B cell epitopes from HN and F proteins of BPIV-3A and BPIV-3C strains. Animal immunization tests show that the antigen peptide and the complex thereof can effectively stimulate the body to produce specific IgG antibodies and neutralizing antibodies, induce Th1 type cellular immune response, and effectively eliminate viruses and reduce lung tissue lesions, and show good immunogenicity and protection effect. The application provides an efficient and safe vaccine candidate for prevention and control of BPIV-3, and has a good application prospect.

The invention discloses a fusion protein for diagnosing mycoplasma bovis infection as well as a preparation method and application of the fusion protein. According to the fusion protein disclosed by the invention, dominant B cell epitope peptides of two strong immunogenic lipoproteins of mycoplasma bovis are connected, mycoplasma bovis infection can be effectively distinguished, and the fusion protein is further used for preparing a detection product for diagnosing mycoplasma bovis infection. A detection product prepared from the fusion protein disclosed by the invention has excellent sensitivity and specificity, and is of great significance for improving the economic benefit of the cattle breeding industry.

The invention relates to the technical field of veterinary drugs, and particularly provides a polynucleotide molecule composition for preventing cat infectious peritonitis, a chimeric polynucleotide molecule and an mRNA vaccine. In order to overcome the ADE effect caused by S protein full-length antigen in the prior art, FIPV N protein without ADE risk is adopted and is combined with screened S protein specific T cell and B cell epitopes. Core schemes include two classes: one is a separate polynucleotide composition comprising epitope peptides encoding N and S proteins; and 2, a single chimeric polynucleotide encoding N protein and S protein epitope peptides. The polynucleotides are optimized by codons and encapsulated in lipid nanoparticles (LNP) to prepare vaccines. Experiments prove that the vaccine can synergistically stimulate powerful immune response, the survival rate of immune cats is remarkably increased to 80%-100% from standard 40% on the premise that ADE is not caused, and FIP is effectively prevented.

The invention belongs to the technical field of biology, and discloses a recombinant turkey herpesvirus, and a tandem epitope expression cassette is inserted between UL45 and UL46 of the recombinant turkey herpesvirus; the tandem epitope expression cassette is used for expressing a plurality of B cell epitopes and a plurality of T cell epitopes. According to the recombinant herpesvirus of turkeys, a tandem epitope is inserted into an HVT (herpesvirus of turkeys) vector, and experiments prove that the recombinant herpesvirus of turkeys can induce higher HI antibody and neutralizing antibody titer, obviously stimulate spleen T lymphocyte response of immunized chicken, and can generate an obvious challenge protection effect in 3 days; the challenge protection effect is obviously superior to that of a combined immunization scheme of H9N2 AIV multi-epitope recombinant baculovirus (BV-BNT) and InV provided by the applicant; meanwhile, the recombinant turkey herpesvirus is verified to be capable of performing intraembryonic vaccination and generating an obvious immune protection effect. Meanwhile, the invention further provides a preparation method and application of the recombinant turkey herpesvirus.

The invention discloses a camel source single-domain antibody CB3 for recognizing linear B cell epitope in clostridium difficile methylase CamA and application of the camel source single-domain antibody CB3. The nucleotide sequence of the camel source single-domain antibody CB3 is as shown in SEQ ID NO: 1, and the amino acid sequence of the camel source single-domain antibody CB3 is as shown in SEQ ID NO: 2. The camel source single-domain antibody CB3 disclosed by the invention has the advantages of strong specificity, high sensitivity, good accuracy and the like, and it is proved that the camel source single-domain antibody CB3 can be used for detecting recombinant expressed CD-CamA and endogenous CD-CamA by methods such as enzyme-linked immunosorbent assay (ELISA) and western blot hybridization (WB).

The invention discloses a B cell epitope peptide based on human papilloma virus E6 protein and application of the B cell epitope peptide in cancer diagnosis. According to the invention, the epitope peptide is displayed and expressed on the surface of an inert carrier bacterium S9H, so that an RI5 epitope peptide-antibody direct mediated agglutination test detection system based on thallus surface epitope display is successfully constructed. Compared with an immunoserological detection technology based on a complete protein antigen, the kit provided by the invention can be used for more specifically, sensitively, conveniently and quickly detecting the virus oncoprotein specific antibody in the body of an HPV persistent infection patient, and can be used for qualitatively and quantitatively determining the content of the antibody. The epitope peptide-antibody direct mediation agglutination test detection technology provides an efficient and reliable new method for accurate diagnosis of dynamic changes of virus oncoprotein antibodies in persistent infection and cancer stages of HPV patients, and has potential important clinical application value.

This invention belongs to the field of bioengineering technology, specifically relating to a Brucella OMP16 B-cell epitope fusion protein and its applications. The fusion protein comprises multiple repeating units linked by flexible linking peptides. Each repeating unit is formed by the tandem sequence of five Brucella OMP16 B-cell epitopes in their native order; the amino acid sequences of the five B-cell epitopes are: TLSKQAQW, LQRYPQY, GQRRAAAT, RDFLASRG, and VPTNRMRT. Based on this, this invention screened for the fusion protein with the best structural stability and theoretical solubility. It strongly binds to Brucella whole-cell polyclonal antibodies and OMP16 polyclonal antibodies, and is specifically recognized by five corresponding OMP16 monoclonal antibodies, indicating that the flexible linking peptides and tandem sequence effectively maintain the native conformation and spatial accessibility of each B-cell epitope.

The application discloses a recombinant virus-like nanoparticle for immunotherapy of gastric cancer and application thereof. The recombinant virus-like nanoparticle is a chimeric recombinant virus-like nanoparticle formed by self-assembly of a fusion protein HBC-CLDN18.2 of a hepatitis B virus core protein and CLDN18.2 tight junction protein. The application uses genetic engineering technology to truncate the C-terminal end of a natural hepatitis B virus core antigen, and mutate cysteine residues at positions 48 and 107 into serine, so that the hepatitis B virus core antigen can self-assemble into a virus-like nanoparticle with strong stability. The virus-like nanoparticle is used as a carrier, a B cell epitope peptide (tight junction protein CLDN18.2) of a gastric cancer tumor-related antigen is inserted into an immunodominant site of the carrier, so that the humoral immune response against gastric cancer is enhanced. Two T cell epitope peptides of the heterologous hepatitis B virus core antigen are used to replace T cell epitope peptides on the carrier, so that the cellular immune response against gastric cancer is enhanced, and strong antitumor effect and immune memory effect are shown.

The application provides an antigen epitope peptide and a paralichthys olivaceus mucin Muc13 specific antibody prepared by the antigen epitope peptide, determines a B cell epitope from the paralichthys olivaceus Muc13 protein through antigen epitope screening, and the sequence is PGTTEEPDPTAGSPP. After the epitope is coupled with a KLH carrier protein, the epitope is used for immunizing New Zealand white rabbits, and a rabbit-derived anti-paralichthys olivaceus Muc13 specific antibody is successfully obtained. The antibody can specifically recognize the Muc13 protein in paralichthys olivaceus mucosa tissue and Muc13 positive mucous cells, and provides a key detection means for in-depth exploration of the function mechanism of mucous cells in the fish mucosa immune response process.

The application belongs to the technical field of animal vaccine preparation, and discloses an African swine fever virus (ASFV) multi-epitope fusion protein, a ferritin nanoparticle vaccine of the ASFV multi-epitope fusion protein and application. The application obtains 16 dominant B cell epitopes by using an ASFV phage display library in combination with reverse screening of clinical positive serum, and constructs a nanoparticle vaccine by linking the 16 epitopes with ferritin self-assembly. The vaccine has excellent biosafety, can induce high-level and sustained specific antibodies and significant IFN-gamma cellular immune response in pigs, can effectively delay the clinical onset and death process of pigs in ASFV virulent challenge experiments, and can significantly reduce the virus load in whole-body organ tissues, thereby providing 25% survival protection rate. The application provides an effective new strategy for the development of a subunit vaccine for responding to various ASFV mutant strains.

The application discloses a monoclonal antibody of a porcine rotavirus VP6 protein, an epitope of the monoclonal antibody and application of the monoclonal antibody. The monoclonal antibody can specifically recognize a linear B cell epitope on the VP6 protein, and a core sequence of the epitope is TTRLSFQLVR. The antibody has high affinity and high specificity, and can cross-react with porcine rotaviruses of various genotypes such as G5, G9 and G11. The application further includes an indirect ELISA detection kit containing the monoclonal antibody, and application of the linear epitope in detection of porcine rotaviruses, development of vaccines and research on functions of viral proteins. The application provides an important tool and theoretical basis for serological diagnosis, vaccine evaluation and etiological research of porcine rotaviruses.

The invention discloses a Klebsiella pneumoniae antigen epitope chimeric protein as well as preparation and application thereof, and relates to the fields of genetic engineering technologies, vaccines and diagnostic reagents. Amino acid sequences of Klebsiella pneumoniae outer membrane proteins OMPA, OMPN and CusC are analyzed through a computer, two B cell epitopes, one HTL epitope and one CTL epitope are respectively screened from the three outer membrane proteins, a cholera toxin subunit B (CTB) is used as an internal adjuvant, an EAAAK joint is used for connecting the amino acid sequence of the CTB and the B cell epitope of the OMPA, the B cell epitope and the HTL epitope are connected through a GPGPG flexible joint, and the CTL epitope and the CusC epitope are connected through a GMPG flexible joint. CTL inner epitopes are connected through an AAY connector to form a chimeric protein with multiple antigen fragments connected in series. A codon preferred by a pronucleus is selected, and a full-length gene of the chimeric protein is chemically synthesized. The chimeric protein is expressed and purified by using a gene engineering technology, and the chimeric protein has 312 amino acids in total length. The expressed chimeric protein can be used for developing vaccines, antibodies or antigen detection reagents.

This invention concerns immunogenic B cell epitopes of the RPO30 protein of the Lumpy Skin Disease virus isolate from East Java as vaccine candidates in the field of animal health. This invention relates to the identification, characterization, and utilization of immunogenic B cell epitopes derived from the RPO30 protein of the Lumpy Skin Disease virus isolate from East Java. The identified B cell epitopes were selected from the group consisting of SL, D, TSSYESTSSQ, and QGMGGYT, which have the ability to induce specific humoral immune responses against the Lumpy Skin Disease virus. These B cell epitopes can be chemically synthesized or recombinantly expressed and formulated with a veterinary acceptable carrier or pharmaceutical adjuvant to produce an epitope-based vaccine candidate. This invention provides an alternative vaccine candidate that is safer than live attenuated vaccines, has the potential to reduce post-vaccination side effects, and is more specific to local Indonesian isolates.This invention is expected to be used for the prevention of Lumpy Skin Disease in cattle and buffalo.

The application provides an antigen epitope polypeptide and a paralichthys ovatus mucin Muc1 specific antibody prepared by the antigen epitope, a B cell epitope from the paralichthys ovatus Muc1 protein is identified through antigen epitope screening, and the amino acid sequence is SEQ ID NO:1. After the epitope is coupled with a KLH carrier protein, a New Zealand white rabbit is immunized, and a rabbit anti-paralichthys ovatus Muc1 polyclonal antibody is successfully prepared. The antibody can specifically recognize the Muc1 protein in paralichthys ovatus mucosa tissue and positive mucous cells, and provides an important detection tool for studying the function mechanism of mucous cells in fish mucosal immunity.

The application discloses a kind of for preventing Bungowenia virus Polyepitope fusion protein, gene, vaccine and preparation method thereof, belong to biological medicine technical field.The Polyepitope fusion protein for preventing Bungowenia virus provided in the application is obtained in series by E. coli outer membrane anchoring protein ClyA, four MHC I class restricted T cell epitopes, four MHC II class restricted T cell epitopes, two B cell epitopes and BALB / c mouse immunoglobulin Fc fragment.The vaccine of the application is prepared by high-pressure homogenization process to obtain the engineered artificial nanovesicle (E-aBNV) with high-density surface display antigen array.The E-aBNV utilizes itself pathogen-associated molecular pattern to provide "self-adjuvant" effect, can efficiently activate B cell receptor and promote antigen cross-presentation, thereby synergistically inducing high level of neutralizing antibody and Th1-biased cellular immunity, which can effectively block the infection of Bungowenia virus to host cell.

The invention belongs to the technical field of animal vaccine preparation, and discloses an African swine fever virus multi-epitope fusion protein and a ferritin nanoparticle vaccine and application thereof. According to the invention, an ASFV phage display library is combined with clinical positive serum reverse screening to obtain 16 dominant B cell epitopes, and the 16 epitopes are connected in series with an epitope and are self-assembled with ferritin to construct the nanoparticle vaccine. The vaccine has excellent biological safety; a high-level and continuous specific antibody and remarkable IFN-gamma cellular immune response can be induced and generated in a pig body; in an ASFV virulent virus challenge experiment, the vaccine can effectively delay clinical morbidity and death processes of pigs, remarkably reduce virus load in visceral organ tissues of the whole body and provide a survival protection rate of 25%. The invention provides an effective new strategy for research and development of subunit vaccines for coping with ASFV multi-variant strains.

The application discloses a fusion protein PP13138R and application thereof in tuberculosis prevention. Specifically disclosed is a fusion protein comprising HTL, CTL and B cell epitopes in series, PorB, PADRE and RS-09. The application screens 34 epitopes for Mycobacterium tuberculosis, which have good immunogenicity and antigenicity and no toxicity and no sensitization, and have the characteristics of high population coverage and the like. The auxiliary peptide PADRE is further added to improve the immunogenicity of the vaccine, and the PorB and RS-09 are added to endow the vaccine with a targeted delivery function. In-vitro experiments prove that the PP13138R can stimulate human peripheral blood mononuclear cells to produce an immune response, and is an advantage protective antigen. The PP13138R has the advantages of simple preparation method, low cost, high yield and higher safety as a vaccine. The application has great value for the prevention and treatment of active tuberculosis and latent tuberculosis infection.

Provided are a virus peptide-protein subunit combination vaccine based on DNA nanotechnology, and a preparation method therefor and use thereof, which belong to the technical field of biological products. Provided is a virus peptide-protein subunit combination vaccine based on DNA nanotechnology, in which a tetrahedral framework nucleic acid formed by assembly of DNA strands is used as a vector, a protein antigen triggering virus-specific T cell activation is coupled to one edge of the tetrahedral framework nucleic acid, and B cell epitope peptides of a virus are coupled to four vertices of the tetrahedral framework nucleic acid.

The invention provides a design method for improving the population coverage rate of a multi-epitope vaccine, which comprises the following steps: carrying out epitope prediction on B cells and T cells according to candidate proteins to obtain B cell epitopes and T cell epitopes; setting a sliding window for the candidate antigen protein; judging whether the needed epitope exists in the sliding window or not; if the needed epitope exists, shortening the length of the window by one amino acid, and judging whether the specifically bound MHC-I / II allele is reduced or not; if the MHC-1 / II allele is reduced, the window shortening operation is revoked, and overlapping epitopes which are specifically combined with the maximum MHC-1 / II allele are reserved, so that candidate overlapping epitopes are obtained. According to the invention, a three-type epitope maximization strategy based on a sliding window is adopted, and population coverage rate maximization and dual immune recognition are realized in a vaccine construct with a compact structure by recognizing the number of specific binding MHC-I / II alleles and the number of partially overlapped B cell epitope amino acids.

The application relates to a self-adjuvant based on a virus-derived glycopeptide and application thereof in preparation of a vaccine, the self-adjuvant being a varicella-zoster virus (VZV)-derived glycopeptide fragment, which is a B cell epitope domain V50 glycopeptide (from a valine at 50 to an isoleucine at 135, V50-I135) in a glycoprotein E. After fusion expression of the V50 glycopeptide and a novel coronavirus RBD, coupling on the surface of a ferritin-based nanoparticle, a self-adjuvant vaccine is formed, which can significantly enhance the RBD-specific antibody and pseudovirus neutralizing antibody levels induced by the RBD nanoparticle vaccine, and also has a synergistic effect in B cell response and T cell response of systemic and mucosal immunity. The immunoprotective effect of the novel coronavirus RBD nanoparticle vaccine is significantly improved.

The invention discloses a bovine parainfluenza virus 3A and 3C type multi-epitope antigen peptide as well as a compound and application thereof. The amino acid sequences of the multi-epitope antigen peptides BPIV-3A and BPIV-3C are respectively shown as SEQ ID NO: 1 and SEQ ID NO: 3, and the multi-epitope antigen peptides are formed by connecting CTL epitopes, HTL epitopes and B cell epitopes screened from HN and F proteins of BPIV-3A and BPIV-3C strains. Animal immune tests show that the antigen peptide and the compound thereof can effectively stimulate a body to generate a specific IgG antibody and a neutralizing antibody, induce immune response of Th1 type cells, effectively eliminate viruses and relieve lung tissue lesions, and show good immunogenicity and protection effect. The invention provides an efficient and safe vaccine candidate for prevention and control of BPIV-3, and has a good application prospect.

The invention belongs to the technical field of biology, and discloses a recombinant turkey herpesvirus, and a tandem epitope expression cassette is inserted between UL45 and UL46 of the recombinant turkey herpesvirus; the tandem epitope expression cassette is used for expressing a plurality of B cell epitopes and a plurality of T cell epitopes. According to the recombinant turkey herpesvirus, tandem epitopes are inserted into an HVT (turkey herpesvirus) carrier, and through experimental verification, good cellular immune response can be induced and generated in 1-day-old chicks and 18-day-old chick embryos by immunizing the multi-epitope recombinant HVT vaccine for one time, and compared with inactivated vaccines needing two-time immunization, the recombinant turkey herpesvirus can provide earlier and better challenge protection effect. Meanwhile, the recombinant turkey herpesvirus is verified to be capable of performing intraembryonic vaccination and generating an obvious immune protection effect. Meanwhile, the invention further provides a preparation method and application of the recombinant turkey herpesvirus.

The invention provides a specific monoclonal antibody for resisting pseudorabies virus IE180 protein as well as a preparation method and application of the specific monoclonal antibody, and belongs to the technical field of biological medicines. The invention provides a hybridoma cell strain pIE180 mAb for producing a specific monoclonal antibody against the pseudorabies virus IE180 protein, and successfully prepares the monoclonal antibody against the PRV IE180 protein. An overlapping peptide library technology shows that the monoclonal antibody can recognize a new B cell epitope, and the epitope is highly conservative in various classic and mutant isolates, can be used for specific detection of PRV infection and development of diagnostic reagents, provides an important molecular basis for design of subunit vaccines, and has broad application prospects. Important clinical application and market prospects are realized.

A recombinant multi-epitope immunogenic polypeptide derived from *C. bancroftian* and its applications are disclosed. The amino acid sequence of the recombinant multi-epitope immunogenic polypeptide includes a fragment of the *Mycobacterium tuberculosis* 50S ribosomal protein L7 / L12, a cytotoxic T lymphocyte epitope, a helper T lymphocyte epitope, and a linear B cell epitope, formed by tandem linkers. The nucleotide coding sequence of the polypeptide is also disclosed. This invention further discloses a recombinant expression vector containing the nucleic acid molecule, a host cell containing the recombinant expression vector, an immunogenic composition containing the recombinant multi-epitope immunogenic polypeptide, and a method for preparing the recombinant multi-epitope immunogenic polypeptide. Immuninformatics analysis results show that the polypeptide is antigenic, non-sensitizing, and non-toxic; structural prediction, molecular docking, and computer immunosimulation results suggest its application potential as a candidate immunogenic polypeptide related to *C. bancroftian*.

The present application belongs to the technical field of genetic engineering, and particularly relates to a bovine rotavirus group A multi-epitope fusion protein and application thereof. The present application provides a bovine rotavirus group A multi-epitope peptide, fusion protein based on ferritin nanocarriers and application thereof. The multi-epitope peptide is based on bovine rotavirus group A VP4 and VP7 protein sequences, and highly conserved and strong immunogenicity cytotoxic T lymphocyte epitopes, helper T cell epitopes and B cell epitopes are screened by using immunoinformatics technology, and are spliced to form a multi-epitope fusion antigen with good conformational stability. Immunological evaluation shows that the multi-epitope fusion antigen has good antigen specificity and neutralizing activity. Animal experiment results show that the epitope peptide and fusion protein can induce the body to produce high-level neutralizing antibodies against multiple genotypes of BRVA G6, G8 and G10, and significantly improve the broad spectrum and durability of immunoprotection.

The application belongs to the technical field of biology and relates to a broad-spectrum monoclonal antibody for recognizing a linear antigen epitope at the C-terminal of foot-and-mouth disease virus VP2 protein and application thereof. The monoclonal antibody pOTB-1 is successfully constructed, the amino acid sequence of the heavy chain variable region is as shown in SEQ ID No. 6, and the amino acid sequence of the light chain variable region is as shown in SEQ ID No. 7. The biological activity of the antibody is verified by indirect immunofluorescence test, enzyme-linked immunosorbent assay and Western-blot. The antibody exhibits broad-spectrum reactivity to O type and A type FMDV and has strong binding affinity. The antibody recognizes a continuous linear B cell epitope at the C-terminal of the structural protein VP2. The epitope sequence is highly conserved among FMDV O type, A type and Asia1 type, which indicates that there is an inter-type common antigen site in the capsid protein VP2. A competitive ELISA detection method is established based on biotinylated pOTB-1, which provides a tool for the prevention and control of FMDV and serological detection, and lays a foundation for the design of FMDV marker vaccine and the evaluation of immunization effect.

The application belongs to the technical field of biotechnology, and provides a Streptococcus suis type 2 (SS2) multi-epitope inhalation subunit vaccine and application thereof.Four CTL epitopes, seven HTL epitopes and six B cell epitopes are screened from the conserved epitopes of SS2 virulence factors SSU05-1022 and SpaA, and the four CTL epitopes, the seven HTL epitopes and the six B cell epitopes are fused by AAY / GPGPG / EAAAK linkers and introduced into L7 / L12 ribosomal protein as an adjuvant to construct an inhalation subunit vaccine 1022-SpaA V3.The vaccine is intranasally primed and boosted at 60 μg, can improve respiratory mucosa sIgA and serum IgG levels, activate related immune cells, and form a mucosal and systemic double immune mechanism.The survival rate of mice after SS2 multi-strain challenge is up to 100%, and the bacterial load can be reduced.The vaccine is safe and non-toxic, inhalation can save cost, and is suitable for industrial scale production.