30 results about "Oral immunization" patented technology

The invention relates to the technical field of biological genetic engineering, in particular to construction of recombinant lactobacillus plantarum of an African swine fever virus antigen, and the method comprises the following steps: constructing a lactobacillus plantarum shuttle vector containing different ASFV antigen genes; and referring to a genome sequence of a Chinese isolate CADCHN09 of ASFV disclosed in GenBank. The method has the advantages that an inducer is not needed, antigen protein is efficiently expressed, the dendritic cell (DC) binding peptide for enhancing immune cell uptake and processing is subjected to fusion expression, the oral immunization mode is simple, mucosal immunity is stimulated, and ASFV infection is effectively prevented and controlled; probiotics, namely lactobacillus plantarum, which naturally exist in animal intestinal tracts and can be colonized and copied in digestive tracts for a long time are used as a carrier; key epitopes of proteins such as nucleocapsid protein p72, outer envelope protein CD2v, inner envelope protein p54, p22, pE199L and pE248R, which play an important role in adsorption, internalization, endocytosis and membrane fusion of ASFV, are preferably fused with dendritic cell binding peptide and then are displayed on the surface of lactobacillus plantarum.

The invention belongs to the technical field of biological genetic engineering, and discloses a lactococcus lactis carrier oral vaccine for enterotoxigenic escherichia coli F18 fimbriae and application. Four lactococcus lactis antigen surface display strains are prepared and are mixed for oral immunization. The oral immunization mode is convenient to operate, the stress reaction of injection immunization to animals is avoided, meanwhile, the immunization effect is greatly improved, and diarrhea of enterotoxigenic escherichia coli to piglets can be effectively prevented. The escherichia coli vaccine is an oral escherichia coli vaccine, a mouse immune challenge experiment shows that the vaccine can be completely survived under the challenge dosage of LD50, a piglet immune challenge experiment shows that the diarrhea condition of piglets can be completely protected, and it is indicated that an immune group generates effective mucosal immune protection capacity.

The invention provides a novel ecological feed capable of preventing multiple grass-bream diseases, the novel ecological feed comprises a feed matrix and an ecological feed leavening agent, and the ecological feed leavening agent comprises probiotics with surface expression antigens, the antigen is any one or more of a grass carp reovirus antigen, a flavobacterium johnsonii antigen, an aeromonas veronii antigen and a clonorchis sinensis antigen. The novel ecological feed with a disease-resistant function in a parabramis pekinensis breeding mode is prepared for the first time, the resistance of the parabramis pekinensis to various pathogens can be enhanced, and the survival rate of the parabramis pekinensis in the disease attack peak period can be increased by 50% or above after immunization. An oral immunization mode is adopted and can be completed in the feeding process, and the immunization and breeding cost is greatly saved.

The invention relates to the field of veterinary biological products, in particular to a porcine epidemic diarrhea virus attenuated strain and a vaccine and application thereof. The porcine epidemic diarrhea virus attenuated strain provided by the invention is obtained through in-vitro separation culture and continuous five rounds of plaque purification and screening technology, and has stable culture characteristics. Meanwhile, the virulence of the strain on newborn piglets and weaned piglets is remarkably reduced, and the virulence does not have the phenomenon of reversion after continuous passage for five times in a pig body. The strain has good immunogenicity, and mucosal immunity and systemic immune response of an organism can be activated at the same time by adopting an oral immunization mode. A neutralizing antibody of 1: 8-1: 16 can be detected in oral fluid and serum after the piglets are immunized for 14 days, and a large amount of sIgA is generated in milk after the pregnant sows are immunized, so that the piglets obtain effective passive immune protection.

The present application relates to the technical field of biological agents, and discloses a kind of M cell targeted peptide and chitosan modified dendritic mesoporous silica oral adjuvant and application thereof.The oral adjuvant includes dendritic mesoporous silica (DFNSs), and M cell targeted peptide (CKS9) modified chitosan (CS), the dendritic mesoporous silica is mixed with M cell targeted peptide modified chitosan at mass ratio 4-6:1, and is made into CKS9-CS-DFNSs oral nano adjuvant.The CKS9-CS-DFNSs oral nano adjuvant provided in the present application has good stability and safety, can effectively induce mucosal immune response and humoral immune response by oral immunization, improve the effect of oral vaccine immunization, and can be widely used in the preparation of animal inactivated vaccine, recombinant protein vaccine and polypeptide vaccine.The oral nano adjuvant preparation has low cost, simple process and is easy to mass-produce.

The invention belongs to the technical field of genetically engineered bacteria, and particularly discloses an African swine fever oral vaccine saccharomyces cerevisiae engineered bacterium as well as a construction method and application thereof. A free plasmid system is adopted to transform saccharomyces cerevisiae, and an engineering strain for displaying ASFV p30 / p54 antigens is constructed. The matched plasmid system has the advantages of being simple and convenient to operate and short in construction period, construction and transformation of recombinant plasmids can be completed in a short time, and the research and development efficiency of vaccine candidate strains is remarkably improved. Meanwhile, the free plasmid can be stably inherited and copied in a yeast host, so that the expression stability of a target antigen in a passage process is guaranteed, and large-scale fermentation preparation is facilitated. After oral immunization, the saccharomyces cerevisiae engineering bacterium constructs a system-mucous membrane dual defense barrier in a tested animal pig body, not only induces high-level system IgG, but also induces remarkable sIgA mucous membrane immunity in intestinal tracts, and provides a new idea and technical reserve for research and development of ASFV subunit vaccines.

The application belongs to the technical field of biological medicine, and discloses a method for preparing an avian infectious bursal disease VP2 nanoparticle oral vaccine by using Kluyveromyces marxianus and application thereof. The applicant assembles and prepares a double replicon expression vector suitable for Kluyveromyces marxianus through screening, and the vector achieves the effect of high conversion rate and high expression efficiency in Kluyveromyces marxianus. The double replicon expression vector pGKD32 is shown in SEQ ID NO. 2. The applicant successfully expresses the avian infectious bursal disease VP2 in the form of nanoparticles by using the Kluyveromyces marxianus expression system provided in the application, and provides a new idea for the research and development of the avian infectious bursal disease virus oral immunization strategy.

The application belongs to the technical field of biological gene engineering, and discloses a lactococcus lactis carrier oral vaccine for producing enterotoxigenic escherichia coli F18 pilus type and application. Four lactococcus lactis antigen surface display strains are prepared, and are mixed to perform oral immunization. The oral immunization mode is convenient to operate, avoids the stress reaction caused by injection immunization on animals, and greatly improves the immunization effect, so that the diarrhea caused by enterotoxigenic escherichia coli on piglets can be effectively prevented. 50 The application is an oral escherichia coli vaccine, and mouse immunization challenge experiment shows that all the mice can survive under the challenge dose of LD , and piglet immunization challenge experiment shows that the piglets can be 100% protected from diarrhea, which shows that the immunization group has effective mucosal immune protection ability.

The present invention belongs to the field of genetic engineering technology and specifically relates to an attenuated Pasteurella multocida strain, a method for its construction, and applications. The attenuated Pasteurella multocida strain lacks the arcA and gatA genes of Pasteurella multocida. The arcA gene of Pasteurella multocida is numbered A0R64_00400. The arcA and gatA double-gene mutant strain constructed by the present invention is highly attenuated in ducks and has a certain level of in vivo colonization, demonstrating its potential as an attenuated live vaccine. Oral immunization with this mutant strain does not affect the production performance of ducks and can provide immune protection against a lethal dose of Pasteurella multocida. The present invention provides a new strategy for developing a live attenuated vaccine against Pasteurella multocida or other bacteria.

The invention belongs to the technical field of genetic engineering, and particularly relates to recombinant lactobacillus plantarum for preventing and controlling chicken necrotic enteritis as well as a preparation method and application of the recombinant lactobacillus plantarum. The invention provides an oral vaccine vector capable of co-expressing various key virulence factor antigens (FimB, NetB, CnaA and FBA) of clostridium perfringens, and by fusing dendritic cell (DC) targeting molecules (nano antibody Phi74), the efficiency of recognizing and taking in the vaccine vector by full-time antigen presenting cells is remarkably improved, so that the immunogenicity is enhanced. When the recombinant lactobacillus is used for immunizing animals, mucosal immunity, cellular immunity and humoral immunity reactions can be induced, the immunity of organisms is comprehensively enhanced, and a safe and efficient oral immunization scheme suitable for large-scale breeding is established for treating chicken necrotic enteritis.

The invention provides attenuated salmonella typhimurium and application thereof, and the provided attenuated salmonella typhimurium is prepared by knocking out a relA gene, a manA gene and a sifA gene. According to the three-gene salmonella attenuated strain provided by the invention, an antigen is presented to a host cell through oral immunization, so that an organism is induced to generate immune response. The action mechanism of the salmonella as the live vector is that after carrying bacterial antigens to pass through intestinal epithelium, the bacterial antigens are transferred to immune cells through intestinal mucosa, and the immune response of a host is triggered. Safety detection shows that the attenuated strain constructed by the invention is high in safety, and can be used as a carrier to carry other pathogenic antigens or drugs to an action site so as to play a role of the carrier.

The application belongs to the technical field of biological medicine, and discloses a method for preparing porcine epidemic diarrhea virus (PEDV) epitope nanoparticles by using Kluyveromyces marxianus and application thereof. The applicant assembles and prepares a double replicon expression vector suitable for the Kluyveromyces marxianus through screening, and the vector achieves the effect of high conversion rate and high expression efficiency in the Kluyveromyces marxianus. The double replicon expression vector pGKD32 is shown as SEQ ID NO. 2. The Kluyveromyces marxianus expression system provided by the application is used to successfully express the PEDV epitope in the form of nanoparticles, thereby providing a new idea for the research and development of the PED oral immunization strategy.

The invention discloses a preparation method and application of an oral chicken infectious anemia vaccine. The preparation method comprises the following steps: providing an eimeria vector; genes of VP1 protein and VP2 protein for coding the chicken infectious anemia virus are introduced into the eimeria vector; after oral immunization, the recombinant eimeria can effectively reduce the clinical symptoms and virus load of chicken lean transmission, saves the manpower and material resources of vaccine immunization, is suitable for prevention and control of poultry epidemic diseases in large-scale breeding industry, and has a wide application prospect.

The invention discloses a rabies virus glycoprotein chimeric recombinant vesicular stomatitis virus and application thereof, and belongs to the technical field of biological medicine. According to the invention, glycoproteins of two rabies viruses are assembled into the vesicular stomatitis virus vector, and the vesicular stomatitis virus recombinant rabies virus prepared from the vesicular stomatitis virus vector has high virus titer and is convenient for in-vitro culture; a vector vaccine prepared from the recombinant virus can induce animals to generate strong rabies virus neutralizing antibody response through oral immunization, and provides 100% challenge protection for the immunized animals, so that the vaccine can be used as a convenient and effective mucosal immune candidate vaccine; the vaccine has important significance on immunization of rabies virus susceptible wild animals and stray dogs and cats. The oral vaccine provided by the invention overcomes the defects of intramuscular injection vaccines, and provides a basis for the development of novel rabies virus vaccines.

This invention discloses a recombinant lactic acid bacteria expressing a truncated avian adenovirus Hexon protein, its construction method, and its applications, belonging to the field of biomedical technology. This invention is the first to construct recombinant lactic acid bacteria strains pNZ8149-FAdV4ΔHexon / NZ3900 and pNZ8149-FAdV8bΔHexon / NZ3900 expressing the truncated avian adenovirus Hexon protein. SDS-PAGE and Western blotting analyses showed that the exogenous genes FADV4-ΔHexon and FADV-8bΔHexon were successfully expressed in *Lactococcus lactis* and exhibited reactivity. Oral immunization of one-day-old chicks showed that oral administration of the recombinant lactic acid bacteria induced humoral and cellular immunity, as well as local mucosal immunity. The results indicate that the recombinant constitutive lactic acid bacteria prepared in this invention can enhance the immunity of chicks.

The invention provides a feline coronavirus recombinant adenovirus. The feline coronavirus recombinant adenovirus is constructed by inserting a feline coronavirus S gene into an adenovirus vector. The invention further provides a preparation method and application of the recombinant adenovirus, the recombinant adenovirus is prepared based on the I-type FCoV S protein, mouse and cat immunization experiments verify that the recombinant adenovirus can significantly enhance the intramuscular injection and oral immune efficacy of FCoV, and the SIgA antibody level and cellular immune response effect induced by oral immunization are better. The recombinant adenovirus vaccine can simultaneously stimulate an organism to generate cellular immunity and humoral immunity, and is good in immune effect, good in protectiveness and high in safety.

The present invention belongs to the field of biomedical technology, and discloses a dual-replicon expression vector applicable to Kluyveromyces marxianus and its application in the preparation of ASFV-P54 nanoparticles. Through screening, assembly and preparation, the applicant prepared a dual-replicon expression vector, which achieved high conversion rate and high expression efficiency in Kluyveromyces marxianus. The dual-replicon expression vector pGKD32 is shown as SEQ ID NO.2. By using the Kluyveromyces marxianus expression system provided by the present invention, the applicant successfully expressed the ASFV-P54 protein in the form of nanoparticles, providing a new idea for the research and development of ASFV oral immunization strategy.

The present application belongs to the technical field of genetic engineering, and particularly relates to a recombinant plant lactobacillus for preventing and controlling chicken necrotic enteritis as well as a preparation method and application thereof. The present application provides an oral vaccine carrier capable of co-expressing multiple key virulence factor antigens (FimB, NetB, CnaA, FBA) of Clostridium perfringens, and the efficiency of the vaccine carrier being recognized and taken by professional antigen presenting cells is significantly improved through fusion of dendritic cell (DC) targeting molecules (nanobody Phi74), so as to enhance immunogenicity. Immunization of animals using the recombinant lactobacillus can induce mucosal immunity, cellular immunity and humoral immunity response, and comprehensively enhance the immune ability of the body, thereby establishing a safe, efficient and suitable oral immunization scheme for large-scale breeding for treating chicken necrotic enteritis.

Immunogenic formulations by oral route comprising: i) one or more chimeric proteins consisting of an immunologically associated antigen from a pathogenic microorganism of the pig and a molecular adjuvant CD154 from the pig; ii) an adjuvant selected from the cyclic dinucleotide monophosphate family; and iii) an excipient acceptable for oral immunization. A method of inducing an immune response against a pathogen characterized in that an immunogenic formulation comprising: i) one or more chimeric proteins formed from an immunologically related antigen from a swine pathogen fused with a molecular adjuvant CD154 from the swine is administered to the swine by oral route; and ii) an adjuvant selected from the cyclic dinucleotide monophosphate family.

The application discloses an oral immune system with a cascade response characteristic as well as a preparation method and application thereof, and belongs to the technical field of biological medicine and nano-preparation. The oral immune system comprises, from inside to outside, a core layer, an intermediate modification layer, a functional unit and a protective shell. The core layer is a live rumen coccus outer membrane vesicle. The intermediate modification layer is a polydopamine coating layer coated on the surface of the core layer. The functional unit is a cerium nanoscale enzyme covalently connected to the surface of the polydopamine coating layer through a thio-ketal bond connecting arm. The protective shell is made of calcium carbonate. The oral immune system integrates oral delivery barrier breakthrough, inflammation microenvironment responsiveness antioxidant and pathogenic bacteria specific immune clearance in the same platform, can break the vicious cycle of flora imbalance, oxidative stress and barrier damage in cooperation, and reduces the interference on beneficial symbiotic flora.

This invention belongs to the field of biomedical technology and discloses a method for preparing porcine rotavirus VP8 nanoparticles using *Kluyveromyces martensii* and its application. The applicant prepared a double-replicon expression vector suitable for *Kluyveromyces martensii* through screening, assembly, and analysis. This vector achieved high conversion rate and high expression efficiency in *Kluyveromyces martensii*. The double-replicon expression vector pGKD32 is shown in SEQ ID NO.2. Using the *Kluyveromyces martensii* expression system provided by this invention, the applicant successfully expressed porcine rotavirus VP8 in nanoparticle form, providing a new approach for the development of oral immunization strategies for porcine rotavirus.

This invention belongs to the field of biomedical technology and discloses a method for preparing porcine parvovirus VP2 nanoparticles from the auxotrophic Kluyveromyces martensii. The applicant, through screening, assembly, and preparation, developed a double-replicant expression vector suitable for Kluyveromyces martensii. This vector achieved high conversion rate and high expression efficiency in Kluyveromyces martensii. The double-replicant expression vector pGKD32 is shown in SEQ ID NO.2. Using the Kluyveromyces martensii expression system provided by this invention, the applicant successfully expressed porcine parvovirus VP2 in nanoparticle form, providing a new approach for the development of oral immunization strategies for porcine parvovirus.

The utility model relates to a bat oral immunization device which comprises a first container which is of a container structure with the upper end open and the lower end closed, and a vaccine solution or a mixed solution of vaccines and attraction liquid is contained in the first container. The grid structure is circumferentially arranged at the opening of the first container; the second container is of a container structure with the upper end open and the lower end closed, and the second container is arranged above the first container and fixed to the grid structure; an inducing liquid is contained in the second container; and the artificial fountain is arranged in the second container and is used for emitting the luring liquid in a fountain manner so as to luring the bats. According to the utility model, the bat can be immunized by oral administration aiming at the wild bat population.

The invention provides a mucosal immune vaccine based on a porcine parainfluenza virus PIV5 carrier and application of the mucosal immune vaccine in immunization of sows and piglets, and belongs to the technical field of animal vaccines. According to the invention, on the basis of a porcine PIV5 reverse genetic operating system, the S gene of the porcine delta coronavirus (PDCoV) is inserted into a P-M gene spacer region of the PIV5 by using a P-M exogenous gene expression cassette of the PIV5, and a recombinant vaccine capable of expressing the PDCoV S is successfully rescued, and is named as JS17-PDCoV-S. A high-titer neutralizing antibody can be generated after the piglets are immunized by oral administration; after a sow orally takes the JS17-PDCoV-S vaccine, high-level serum IgG and mucous membrane sIgA antibodies can be generated. Meanwhile, the piglets are passively immunized by sucking milk, whole body and mucous membrane immune protection aiming at specific pathogens is provided for the piglets, early infection is effectively prevented, and the problem that IgG is easily degraded by digestive tracts of the piglets is solved.

The invention provides an oral vaccine based on a porcine parainfluenza virus PIV5 carrier as well as a preparation method and application of the oral vaccine, and belongs to the technical field of viral vectors and animal vaccines. According to the invention, pig source PIV5 and monkey source PIV5 are respectively taken as carriers, S protein of porcine delta coronavirus (PDCoV) is taken as an antigen, and monkey source PIV5 recombinant virus and pig source PIV5 recombinant virus for expressing PDCoV-S are respectively constructed. Piglet immunization is carried out in an oral-oral immunization mode, the antibody level after immunization is detected, and the result shows that the level of a neutralizing antibody induced by the pig source PIV5 recombinant virus is higher than that of a monkey source PIV5 recombinant virus. The ELISA (enzyme-linked immunosorbent assay) result proves that the IgG (immunoglobulin G) level mediated by the porcine PIV5 recombinant virus is higher than the IgG level mediated by the monkey PIV5 recombinant virus, which indicates that the PIV5-JS17 strain is more suitable for serving as an exogenous gene delivery system of pigs than the PIV5-W3A strain.

The invention discloses a novel ecological feed with a disease-resistant function in a perch-siniperca chuatsi breeding mode, the feed comprises a feed matrix and an ecological feed leavening agent, and the ecological feed leavening agent comprises at least two of lactococcus lactis, bacillus subtilis and pichia pastoris. The novel ecological feed with the disease-resistant function in the persiniperca chuatsi breeding mode is prepared for the first time, the resistance of persiniperca chuatsi to various pathogens can be enhanced, and the survival rate of persiniperca chuatsi breeding in the disease attack peak period can be increased by 50% or above after immunization. An oral immunization mode is adopted and can be completed in the feeding process, and the immunization and breeding cost is greatly saved.

The invention discloses a preparation method and application of salmonella choleraesuis bionic vesicles capable of being used for oral delivery of double antigens, and relates to the field of vaccines.Lipid A of a salmonella choleraesuis attenuated strain SC014 is modified through a genetic engineering technology, low-endotoxin salmonella choleraesuis engineering bacteria are obtained, then SpyCatcher and SpG are expressed on the surfaces of the salmonella choleraesuis engineering bacteria at the same time through ClyA, and the salmonella choleraesuis bionic vesicles are obtained. According to the present invention, the subunit vaccine model GDH-SpyTag and gD-Fc of the clinically common mixed infection pathogen Streptococcus suis and Pseudorabies virus (PRV) can be respectively coupled, and the negative electricity characteristic of the surface of the subunit vaccine model GDH-SpyTag and gD-Fc are used to wrap the subunit vaccine model GDH-SpyTag and gD-Fc by using the cationic compound COS so as to protect the antigen from being damaged by the gastrointestinal tract environment after the oral immunization; and the presentation of the antigen to digestive tract mucosa immune cells is promoted.

This invention belongs to the field of biomedical technology and discloses a method for efficiently and stably expressing canine parvovirus VP2 nanoparticles in *Kluyveromyces martensii*. The applicant prepared a double-replicant expression vector suitable for *Kluyveromyces martensii* through screening, assembly, and analysis. This vector achieved high conversion rate and high expression efficiency in *Kluyveromyces martensii*. The double-replicant expression vector pGKD32 is shown in SEQ ID NO.2. Using the *Kluyveromyces martensii* expression system provided by this invention, the applicant successfully expressed canine parvovirus VP2 in nanoparticle form, providing a new approach for the development of oral immunization strategies for canine parvovirus.

The invention provides a novel multi-connected ecological feed with a disease-resistant function in a carp and crucian carp culture mode, which comprises an ecological feed leavening agent, and the ecological feed leavening agent comprises probiotics of which the surfaces display or secrete expression antigens, the antigen is any one or more of a crucian herpesvirus CaHV antigen, a cyprinid herpesvirus type 2 CyHV2 antigen, a covert death nodavirus CMNV antigen, a carp edema virus CEV antigen and an aeromonas veronii antigen. The novel multi-connected ecological feed for carps and crucians is prepared for the first time, the resistance of carps and crucians to co-infection pathogens can be improved, and the survival rate of carps and crucians can be increased after immunization. An oral immunization mode is adopted and can be completed in the feeding process, and the immunization and breeding cost is greatly saved.

The application discloses yheS The application relates to the field of genetic engineering, and relates to application of a gene in regulation of Riemerella anatipestifer virulence and immunogenicity. ΔyheS The application constructs a gene deletion strain and a gene back-supplementation strain C ΔyheS The results show that yheS Gene deletion can significantly affect growth of the Riemerella anatipestifer, especially under iron limitation conditions; yheS After deletion, the RA bacterial morphology becomes more elongated. yheS Gene deletion leads to significant decrease of virulence of the Riemerella anatipestifer, and the bacterial load in liver, spleen and brain is decreased. yheS The gene deletion strain can realize 100% protection of an animal body through intramuscular injection immunization and / or oral immunization. The above results all show that yheS yheS The gene is an important virulence factor of the Riemerella anatipestifer, and the gene deletion strain can be used as an ideal selection of a new live vaccine of the Riemerella anatipestifer.